LOCUS pTRIX2-Luc::Neon-Hyg 9702 bp ds-DNA circular SYN 25-AUG-2017 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS . SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 9702) AUTHORS MCT TITLE Direct Submission JOURNAL Exported Jan 18, 2018 from SnapGene Viewer 4.0.6 http://www.snapgene.com FEATURES Location/Qualifiers source 1..9702 /organism="synthetic DNA construct" /mol_type="other DNA" rep_origin complement(6..461) /direction=LEFT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind 603..619 /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" promoter 626..644 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" promoter 659..1475 /label=RRNA promoter CDS 1481..3853 /codon_start=1 /label=LUC::NEON fusion gene /translation="MEDAKNIKKGPAPFYPLEDGTAGEQLHKAMKRYALVPGTIAFTDA HIEVNITYAEYFEMSVRLAEAMKRYGLNTNHRIVVCSENSLQFFMPVLGALFIGVAVAP ANDIYNERELLNSMNISQPTVVFVSKKGLQKILNVQKKLPIIQKIIIMDSKTDYQGFQS MYTFVTSHLPPGFNEYDFVPESFDRDKTIALIMNSSGSTGLPKGVALPHRALCVRFSHA RDPIFGNQIAPDTAILSVVPFHHGFGMFTTLGYLICGFRVVLMYRFEEELFLRSLQDYK IQTALLVPTLFSFLAKSTLIDKYDLSNLHEIASGGAPLSKEVGEAVAKGFHLPGIRQGY GLTETTSAILVTPIGDDKPGAVGKVVPFFEAKVVDLDTGKTLGVNQRGELCVRGPMIMS GYVNNPEATNALIDKDGWLHSGDIAYWDEDEHFFIVDRLKSLIKYKGYQVAPAELESIL LQHPNIRDAGVAGLPDDDAGELPAAVVVLEHGKTMTEKEIVDYVASQVTTAKKLRGGVV FVDEVPKGLTGKLDARKIREILIKAKKGGKITAGPGSATVSKGEEDNMASLPATHELHI FGSINGVDFDMVGQGTGNPNDGYEELNLKSTKGDLQFSPWILVPHIGYGFHQYLPYPDG MSPFQAAMVDGSGYQVHRTMQFEDGASLTVNYRYTYEGSHIKGEAQVKGTGFPADGPVM TNSLTAADWCRSKKTYPNDKTIISTFKWSYTTGNGKRYRSTARTTYTFAKPMAANYLKN QPMYVFRKTELKHSKTELNFKEWQKAFTDVMGMDELYK" CDS 4606..5631 /codon_start=1 /gene="aph(4)-Ia" /product="aminoglycoside phosphotransferase from E. coli" /label=HygR /note="confers resistance to hygromycin" /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK E" misc_feature 6409..7490 /label=3' RRNA target promoter complement(7517..7535) /label=T3 promoter /note="promoter for bacteriophage T3 RNA polymerase" primer_bind complement(7556..7572) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" protein_bind 7580..7596 /label=lac operator /bound_moiety="lac repressor encoded by lacI" /note="The lac repressor binds to the lac operator to inhibit transcription in E. coli. This inhibition can be relieved by adding lactose or isopropyl-beta-D-thiogalactopyranoside (IPTG)." promoter complement(7604..7634) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 7649..7670 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." rep_origin complement(7958..8546) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" CDS complement(8717..9577) /codon_start=1 /gene="bla" /product="beta-lactamase" /label=AmpR /note="confers resistance to ampicillin, carbenicillin, and related antibiotics" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS LIKHW" promoter complement(9578..9682) /gene="bla" /label=AmpR promoter ORIGIN 1 cacctaaatt gtaagcgtta atattttgtt aaaattcgcg ttaaattttt gttaaatcag 61 ctcatttttt aaccaatagg ccgaaatcgg caaaatccct tataaatcaa aagaatagac 121 cgagataggg ttgagtgttg ttccagtttg gaacaagagt ccactattaa agaacgtgga 181 ctccaacgtc aaagggcgaa aaaccgtcta tcagggcgat ggcccactac gtgaaccatc 241 accctaatca agttttttgg ggtcgaggtg ccgtaaagca ctaaatcgga accctaaagg 301 gagcccccga tttagagctt gacggggaaa gccggcgaac gtggcgagaa aggaagggaa 361 gaaagcgaaa ggagcgggcg ctagggcgct ggcaagtgta gcggtcacgc tgcgcgtaac 421 caccacaccc gccgcgctta atgcgccgct acagggcgcg tcccattcgc cattcaggct 481 gcgcaactgt tgggaagggc gatcggtgcg ggcctcttcg ctattacgcc agctggcgaa 541 agggggatgt gctgcaaggc gattaagttg ggtaacgcca gggttttccc agtcacgacg 601 ttgtaaaacg acggccagtg aattgtaata cgactcacta tagggcgaat tggagctcga 661 cgtcggtgca ccctgggata tttgcgcacc cacctttccc gtcgcaatcg gcgccgtccg 721 aacgcggaaa tgttccgaga ataatacggc tgggctgcgc cgcgcgttcc ccgcgtgaac 781 caactattgt gcgatgtgca gcatcctatt gtggtgcttt accgcacacc agtttccttg 841 cggggcttat ttggtgtgtt ggatttttaa ttttttgatg attcggagga attttttgca 901 atttttgggt acccctaaaa aaattttgga gtttccgggg ttcgatccaa cttgtgcttt 961 tttaaccgtc catttgtgtc tagtacatca tggagcggta ttctctccgg tggcccaggt 1021 ggcgtcggaa ggaatcggtg tccctcagga tgtttggttt ttcgtttttt tttttgcggt 1081 cttttgtttc gcatcgccgt atcgtgtgcg gcgcgacctt ctttttggct cattggtcag 1141 tgaggtttat aataaagtgg atcatttggg tttttggccc tttgcggcgc acccgcggaa 1201 tgggggaggg attttcatat aattgtagtg tacacattgt gtgtgggtcg caaaatgtca 1261 tgccgtgtgc aatgtttatt tgcgtgctgt gcgtgtcctt ttgtgattcc accacttgtg 1321 tgtattttac cttatcgtct ctccctgccg ttggctgtgc gtgaattgtg cttcgcattc 1381 gtgggaaatc aaaaggggaa tcaacgtgtt cttttttttt cagaagggga gtcaaagcgc 1441 cgtgtggatg ccaagtctgt gcaagagttc tagtggatcc atggaggacg ccaagaacat 1501 caagaaggga ccagccccct tctaccccct ggaggacggc acagccggcg agcagctgca 1561 caaggccatg aagcggtacg ccctggtgcc aggcaccatc gccttcaccg acgcccacat 1621 cgaggtgaac atcacctacg ccgagtactt cgagatgagc gtgcggctgg ccgaggccat 1681 gaagcggtac ggcctgaaca ccaaccaccg gatcgtggtg tgcagcgaga acagcctgca 1741 gttcttcatg cccgtgctgg gagccctgtt catcggcgtg gccgtggccc cagccaacga 1801 catctacaac gagcgggagc tgctgaacag catgaacatc agccagccca ccgtggtgtt 1861 cgtgagcaag aagggcctgc agaagatcct gaatgtgcag aagaagctgc ccatcatcca 1921 gaagatcatc atcatggaca gcaagaccga ttaccagggc ttccagagca tgtacacctt 1981 cgtgaccagc cacctgcccc caggcttcaa cgagtacgac ttcgtgcccg agagcttcga 2041 ccgggacaag accatcgccc tgatcatgaa cagcagcggc agcaccggcc tgcccaaggg 2101 cgtggccctg ccccaccggg ccctgtgcgt gcggttcagc cacgccagag accccatctt 2161 cggcaaccag atcgcccccg acaccgccat cctgagcgtg gtgcccttcc accacggctt 2221 cggcatgttc accaccctgg gctacctgat ctgcggcttc cgggtggtgc tgatgtacag 2281 gttcgaggag gagctgttcc tgcggagcct gcaggactac aagatccaga ccgccctgct 2341 ggtgcccacc ctgttcagct tcctggccaa gagcaccctg atcgacaagt acgacctgag 2401 caacctgcac gagatcgcct ctggcggagc cccactgagc aaggaggtgg gcgaggccgt 2461 ggccaagggc ttccacctgc caggcatccg gcagggctac ggcctgaccg agaccaccag 2521 cgccatcctg gtgaccccca tcggcgacga caagcccgga gccgtgggca aggtggtgcc 2581 cttcttcgag gccaaggtgg tggacctgga caccggcaag accctgggcg tgaaccagag 2641 aggcgagctg tgcgtgagag gccccatgat catgagcggc tacgtgaaca accccgaggc 2701 caccaacgcc ctgatcgaca aggacggctg gctgcacagc ggcgacatcg cctactggga 2761 cgaggacgag cacttcttca tcgtggaccg gctgaagagc ctgatcaagt acaagggcta 2821 ccaggtggcc ccagccgagc tggagagcat cctgctgcag caccccaaca tccgggacgc 2881 cggagtggcc ggactgcccg acgacgacgc cggagagctg ccagccgccg tggtggtgct 2941 ggagcacggc aagaccatga ccgagaagga gatcgtggac tacgtggcca gccaggtgac 3001 caccgccaag aagctgagag gcggcgtggt gttcgtggac gaggtgccca agggcctgac 3061 cggcaagctg gacgccagaa agatccggga gatcctgatc aaggccaaga agggcggcaa 3121 gatcacggcg ggcccgggct cggcgacggt ctcgaaaggt gaggaagaca atatggccag 3181 cttgccagca acgcacgagc ttcatatatt tgggtcgatc aacggtgtgg atttcgacat 3241 ggtgggtcaa ggcacaggta atcctaatga cgggtatgaa gaactcaatt tgaagagcac 3301 taagggagat ctccagtttt ccccgtggat tcttgtgccg catattggtt atggttttca 3361 tcagtactta ccataccccg atggaatgtc gcccttccaa gcagctatgg tggacggctc 3421 ggggtatcaa gtacatcgca ccatgcaatt tgaggacgga gcatccctca ccgtaaacta 3481 ccgctatacg tatgagggga gccacataaa aggtgaggct caagttaaag gcacaggctt 3541 cccagctgac ggcccggtca tgaccaattc attaaccgct gctgattggt gccgttctaa 3601 gaagacctac cctaacgaca agacgatcat tagtactttt aagtggagtt ataccacagg 3661 aaacggaaag cggtatcgaa gcactgctcg aacaacctac acatttgcga agccaatggc 3721 cgcgaactac ttgaagaacc agccaatgta tgtgtttcga aaaacggaac tgaagcattc 3781 gaaaaccgag cttaatttta aagagtggca gaaggcgttt acagacgtga tggggatgga 3841 cgaattgtat aagtgactcg agccatttac gactccaagg caacgctgca gaacaacctg 3901 ccgaaggagc gccgcttctt caagattgtg tcgtggtacg acaacgagtg gggatactcc 3961 caccgcgtgg tggaccttgt acgccacatg gcctcgaagg atcgttcggc aaggttgtag 4021 gcgtggcgat gacttcaggt ctttcttttg cgaataggga tcttataata cacgatgcgt 4081 gtcccgtgat gatcgttacc ggtgctgcca cgatccaatt gacacagcgt caagagcaaa 4141 acaattttac ttttcccttt aaggacaaca acaaaaaaat atataacttt tttttctttt 4201 tttttttttt ttgaaattat atttatggtc atctttggga aacaaaaagc agcaatttaa 4261 tgatgcggaa ggatgagtga aataatgttt aatcaatgta cgaggatttg gggtattgca 4321 aggaaaatgt agatgattta attgggtgtg tgatgcagct tgtggtaatt tttgctcact 4381 tccctttttg ccacatcttt ttagtttttc tgcttttctt tccccattat tccacttgtc 4441 tctcttttcc cacgtttcct gcacgaatgc agaaagtgat atttttactt tgaaagccat 4501 ctaccaacaa caattacatt gaacagaatt tgggattgcg aattaattcc tgcagccccg 4561 cgtgtggcct cgaacaccga gcgaccctgc agccaatgcg gccgcatgaa aaagcctgaa 4621 ctcaccgcga cgtctgtcga gaagtttctg atcgaaaagt tcgacagcgt ctccgacctg 4681 atgcagctct cggagggcga agaatctcgt gctttcagct tcgatgtagg agggcgtgga 4741 tatgtcctgc gggtaaatag ctgcgccgat ggtttctaca aagatcgtta tgtttatcgg 4801 cactttgcat cggccgcgct cccgattccg gaagtgcttg acattgggga attcagcgag 4861 agcctgacct attgcatctc ccgccgtgca cagggtgtca cgttgcaaga cctgcctgaa 4921 accgaactgc ccgctgttct gcagccggtc gcggaggcca tggatgcgat cgctgcggcc 4981 gatcttagcc agacgagcgg gttcggccca ttcggaccgc aaggaatcgg tcaatacact 5041 acatggcgtg atttcatatg cgcgattgct gatccccatg tgtatcactg gcaaactgtg 5101 atggacgaca ccgtcagtgc gtccgtcgcg caggctctcg atgagctgat gctttgggcc 5161 gaggactgcc ccgaagtccg gcacctcgtg cacgcggatt tcggctccaa caatgtcctg 5221 acggacaatg gccgcataac agcggtcatt gactggagcg aggcgatgtt cggggattcc 5281 caatacgagg tcgccaacat cttcttctgg aggccgtggt tggcttgtat ggagcagcag 5341 acgcgctact tcgagcggag gcatccggag cttgcaggat cgccgcggct ccgggcgtat 5401 atgctccgca ttggtcttga ccaactctat cagagcttgg ttgacggcaa tttcgatgat 5461 gcagcttggg cgcagggtcg atgcgacgca atcgtccgat ccggagccgg gactgtcggg 5521 cgtacacaaa tcgcccgcag aagcgcggcc gtctggaccg atggctgtgt agaagtactc 5581 gccgatagtg gaaaccgacg ccccagcact cgtccgaggg caaaggaata gaagcttgtc 5641 gactgagggg atcgatccgg aacaacctgc cgaaggagcg ccgcttcttc aagattgtgt 5701 cgtggtacga caacgagtgg ggatactccc accgcgtggt ggaccttgta cgccacatgg 5761 cctcgaagga tcgttcggca aggttgtagg cgtggcgatg acttcaggtc tttcttttgc 5821 gaatagggat cttataatac acgatgcgtg tcccgtgatg atcgttaccg gtgctgccac 5881 gatccaagtg acacagtgtc aagagcaaaa caattttact tttcccttta aggacaacaa 5941 caaaaaaata tataactttt ttttcttttt ttttttttga aattatattt atggtcatct 6001 ttgggaaaca aaaagcagca atttaatgat gcggaaggat gagtgaaata atgtttaatc 6061 aatgtacgag gatttggggt attgcaagga aaatgtagat gatttagttg ggtgtatgat 6121 gcagcttgtg gtaatttttt acaactttgc gttattggaa gattttggtt gttcagtttc 6181 tttttgttta taactgactg ttttaattta gtgtgatttt ttttttttaa ggatttaagc 6241 tgtttttctt tttttttctt tttcttcact gatggtgttt aaggtgtagt ttagtgtttg 6301 tgcattatgc gtcgtagtgt tgttgttctt tggacttttt cgaacaatac tgggcattat 6361 agcccacctg tgtatatgcc gctagagtat gctagccgtg tgggtaccta ttattactac 6421 tgctgctgct gctgccactc ctacatatgc gcacaagtac actatatata tgcgaataca 6481 cctgtgtgta ttatgctatg tgtgtgtctg tgtgggtgtg taggtgatgg ggtagtggtt 6541 gtattatagt atagtatata tagtagtata cggcaaaagg caaagacaca agcacggcga 6601 gtacgacgaa caaacccatt tcacacatga gtgtgcgcca cacacgcgac tgctcctttt 6661 ttttctgcct tgccaccacc ccatatgtat attatatata tatatatata agggtgtgtg 6721 tgtggtgaga gaaaatagga aatgattgcg tgtgtgtcgc cgccacgaac ctgtcagatg 6781 ctttcttacg gtattttgtt gcatgcaccc gcgtacacac aatatataca cacacaaaaa 6841 aaaacaccga tatgatggcg agtgtgcgat tctcctttta ttgtcttttt cctctttttc 6901 tgtgctctgc tgtgtgccaa taaaacacac gagagtagag aaacggggga aaaagtagta 6961 aataaatgga ggactgcttc gcagctgccc tgccgcactg tttttgctgc tgggttcgca 7021 acgcgatgct gttgagagtg aaatcacatc acaaaaagca cccctcagac tggtgtgaag 7081 tgtgaagtat gtgtacaaat gcgtgtgtgt gtgtatacac attgacgaat agcataccag 7141 ggaaaaaagg gaggacactc tctttcgtgc gtgctgaact ctgtgtgtgt gtgctgccgt 7201 tttgtttgca gcttgaaatg cttggttgaa tagaaatcgg tacttgattg ttgccgcatt 7261 tcgaggaatt tcatccaaaa aaaaaaagag gtcaacaaca accaactgac aaactttgtg 7321 ttggacctgt gtatgtggtg gtgtggtggg ggaagagaaa cacacacaca gacaaacaca 7381 aacaccatgt cattttggca tgcgtgtgcg tgcgtgcgtg cgttgaatat cctccacaca 7441 caaatatata tatataataa taccacaaca acatttcaaa ggatcatcaa ggcgcgccgg 7501 tacccagctt ttgttccctt tagtgagggt taatttcgag cttggcgtaa tcatggtcat 7561 agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa 7621 gcataaagtg taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc 7681 gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc 7741 aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact 7801 cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac 7861 ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa 7921 aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg 7981 acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa 8041 gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc 8101 ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac 8161 gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac 8221 cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg 8281 taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt 8341 atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagga 8401 cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct 8461 cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga 8521 ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg 8581 ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct 8641 tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt 8701 aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc 8761 tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg 8821 gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag 8881 atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt 8941 tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag 9001 ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt 9061 ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca 9121 tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg 9181 ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat 9241 ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta 9301 tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca 9361 gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct 9421 taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat 9481 cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa 9541 agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt 9601 gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa 9661 ataaacaaat aggggttccg cgcacatttc cccgaaaagt gc // LOCUS pLEW CAS9 13009 bp ds-DNA circular SYN 08-DEC-2016 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS . SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 13009) AUTHORS . TITLE Direct Submission JOURNAL Exported Jan 18, 2018 from SnapGene Viewer 4.0.6 http://www.snapgene.com FEATURES Location/Qualifiers source 1..13009 /organism="synthetic DNA construct" /mol_type="other DNA" misc_feature 1..906 /label=B Tub targeting misc_feature 910..1106 /label=ALD SAS /note="aldolase-derived sas from HD330" misc_feature 1107..1198 /label=PARP SAS /note="PARP-derived splice aceptor site from HD108" misc_feature 1236..3923 /label=T7RNAPOL CDS 1278..1298 /codon_start=1 /product="nuclear localization signal of SV40 large T antigen" /label=SV40 NLS /translation="PKKKRKV" primer_bind 2264..2283 /label=Primer 1 misc_feature 3946..4090 /label=ALD 3' UTR /note="truncated aldolase 3' UTR" misc_feature 4094..4205 /label=ACT SAS CDS 4262..5128 /codon_start=1 /label=NEO /translation="MREIVCVQAGQCGNQIGSKFWEVIRPSLDGLHAGSPAAWVERLFG YDWAQQTIGCSDAAVFRLSAQGRPVLFVKTDLSGALNELQDEAARLSWLATTGVPCAAV LDVVTEAGRDWLLLGEVPGQDLLSSHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKH RIERARTRMEAGLVDQDDLDEEHQGLAPAELFARLKARMPDGEDLVVTHGDACLPNIMV ENGRFSGFIDCGRLGVADRYQDIALATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFY RLLDEFF" misc_feature 5148..5435 /label=ACT 3' UTR /note="truncated aldolase 3' UTR" misc_feature 5436..5555 /label=ACT SAS CDS 5584..5649 /codon_start=1 /product="three tandem FLAG(R) epitope tags, followed by an enterokinase cleavage site" /label=3xFLAG /translation="DYKDHDGDYKDHDIDYKDDDDK" CDS 5656..5676 /codon_start=1 /product="nuclear localization signal of SV40 large T antigen" /label=SV40 NLS /translation="PKKKRKV" CDS 5701..9801 /codon_start=1 /product="Cas9 (Csn1) endonuclease from the Streptococcus pyogenes Type II CRISPR/Cas system" /label=Cas9 /note="generates RNA-guided double strand breaks in DNA" /translation="DKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL YYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD ATLIHQSITGLYETRIDLSQLGGD" CDS 9802..9849 /codon_start=1 /product="bipartite nuclear localization signal from nucleoplasmin" /label=nucleoplasmin NLS /translation="KRPAATKKAGQAKKKK" promoter complement(10245..10263) /label=SP6 promoter /note="promoter for bacteriophage SP6 RNA polymerase" promoter 11049..11153 /gene="bla" /label=AmpR promoter CDS 11154..12014 /codon_start=1 /gene="bla" /product="beta-lactamase" /label=AmpR /note="confers resistance to ampicillin, carbenicillin, and related antibiotics" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS LIKHW" rep_origin 12185..12773 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" ORIGIN 1 ctgaccgtat catgatgact ttctccatca tcccatcccc caaggtgtcc gacactgtcg 61 tcgagccgta caatacgact ctctccgtgc accaacttgt ggaaaactcc gatgagtcga 121 tgtgcattga caacgaggca ctgtacgata tttgcttccg caccctgaaa ctgacaacac 181 caacgttcgg tgacctgaac cacttggtgt ctgctgttgt gtccggcgtc acctgctgcc 241 tgcgcttccc tggtcagttg aactctgacc tccgtaagtt ggctgtgaac cttgtcccat 301 tcccgcgtct gcacttcttc atgatgggct tcgccccgct gaccagccgc ggctcgcagc 361 agtaccgcgg tctctccgtg cccgagctaa cgcagcagat gttcgatgcg aaaaacatga 421 tgcaagctgc agatcctcgt cacggccgct acctgacagc gtctgcactc ttccgcggcc 481 gcatgtcgac gaaggaggtt gatgagcaga tgctgaacgt gcagaacaag aactcgtcct 541 acttcattga gtggatcgat cccgaacaac atcaagtcct ctgtttgcga tatcccaccc 601 aagggactca agatggctgt caccttcatt ggcaacaaca cctgcatcca ggagatgttc 661 cgccgtgtgg gagagcagtt caccctcatg ttccgtcgca aggcgttctt gcactggtac 721 actggcgagg gtatggacga gatggaattc acggaggcag agtccaacat gaacgatctc 781 gtgtctgagt accagcagta ccaggatgcc acgattgagg aggagggcga gttcgacgag 841 gaggagcaat actagacgcg gacggggcat ttcccgttcg tcattagcag taggtaatga 901 agatgctcga gggtgctcaa gctgtgtagc gcacgcgttt ccttacatat ttctctaaca 961 ggcacggaag cctaacaaat acacttggct tatttttttg ccccctcatg tcttgtacaa 1021 atatttgcga tagcttagct atcagccaca ttaatcaaac aagtatacca acaagcccga 1081 aaacataaac tcaactgcaa cgaagctggg ctgcacgcgc cttcgagttt tttttccttt 1141 tccccatttt tttcaacttg aagacttcaa ttacaccaaa aagtaaaatt cacaagctga 1201 tctgatccgg atttactaac tggaagaggc actaaatgaa cacgattaac atcgctaaga 1261 acgaattcct cgagcctcca aaaaagaaga gaaaggtcga attctctgac atcgaactgg 1321 ctgctatccc gttcaacact ctggctgacc attacggtga gcgtttagct cgcgaacagt 1381 tggcccttga gcatgagtct tacgagatgg gtgaagcacg cttccgcaag atgtttgagc 1441 gtcaacttaa agctggtgag gttgcggata acgctgccgc caagcctctc atcactaccc 1501 tactccctaa gatgattgca cgcatcaacg actggtttga ggaagtgaaa gctaagcgcg 1561 gcaagcgccc gacagccttc cagttcctgc aagaaatcaa gccggaagcc gtagcgtaca 1621 tcaccattaa gaccactctg gcttgcctaa ccagtgctga caatacaacc gttcaggctg 1681 tagcaagcgc aatcggtcgg gccattgagg acgaggctcg cttcggtcgt atccgtgacc 1741 ttgaagctaa gcacttcaag aaaaacgttg aggaacaact caacaagcgc gtagggcacg 1801 tctacaagaa agcatttatg caagttgtcg aggctgacat gctctctaag ggtctactcg 1861 gtggcgaggc gtggtcttcg tggcataagg aagactctat tcatgtagga gtacgctgca 1921 tcgagatgct cattgagtca accggaatgg ttagcttaca ccgccaaaat gctggcgtag 1981 taggtcaaga ctctgagact atcgaactcg cacctgaata cgctgaggct atcgcaaccc 2041 gtgcaggtgc gctggctggc atctctccga tgttccaacc ttgcgtagtt cctcctaagc 2101 cgtggactgg cattactggt ggtggctatt gggctaacgg tcgtcgtcct ctggcgctgg 2161 tgcgtactca cagtaagaaa gcactgatgc gctacgaaga cgtttacatg cctgaggtgt 2221 acaaagcgat taacattgcg caaaacaccg catggaaaat caacaagaaa gtcctagcgg 2281 tcgccaacgt aatcaccaag tggaagcatt gtccggtcga ggacatccct gcgattgagc 2341 gtgaagaact cccgatgaaa ccggaagaca tcgacatgaa tcctgaggct ctcaccgcgt 2401 ggaaacgtgc tgccgctgct gtgtaccgca aggacaaggc tcgcaagtct cgccgtatca 2461 gccttgagtt catgcttgag caagccaata agtttgctaa ccataaggcc atctggttcc 2521 cttacaacat ggactggcgc ggtcgtgttt acgctgtgtc aatgttcaac ccgcaaggta 2581 acgatatgac caaaggactg cttacgctgg cgaaaggtaa accaatcggt aaggaaggtt 2641 actactggct gaaaatccac ggtgcaaact gtgcgggtgt cgataaggtt ccgttccctg 2701 agcgcatcaa gttcattgag gaaaaccacg agaacatcat ggcttgcgct aagtctccac 2761 tggagaacac ttggtgggct gagcaagatt ctccgttctg cttccttgcg ttctgctttg 2821 agtacgctgg ggtacagcac cacggcctga gctataactg ctcccttccg ctggcgtttg 2881 acgggtcttg ctctggcatc cagcacttct ccgcgatgct ccgagatgag gtaggtggtc 2941 gcgcggttaa cttgcttcct agtgaaaccg ttcaggacat ctacgggatt gttgctaaga 3001 aagtcaacga gattctacaa gcagacgcaa tcaatgggac cgataacgaa gtagttaccg 3061 tgaccgatga gaacactggt gaaatctctg agaaagtcaa gctgggcact aaggcactgg 3121 ctggtcaatg gctggcttac ggtgttactc gcagtgtgac taagcgttca gtcatgacgc 3181 tggcttacgg gtccaaagag ttcggcttcc gtcaacaagt gctggaagat accattcagc 3241 cagctattga ttccggcaag ggtctgatgt tcactcagcc gaatcaggct gctggataca 3301 tggctaagct gatttgggaa tctgtgagcg tgacggtggt agctgcggtt gaagcaatga 3361 actggcttaa gtctgctgct aagctgctgg ctgctgaggt caaagataag aagactggag 3421 agattcttcg caagcgttgc gctgtgcatt gggtaactcc tgatggtttc cctgtgtggc 3481 aggaatacaa gaagcctatt cagacgcgct tgaacctgat gttcctcggt cagttccgct 3541 tacagcctac cattaacacc aacaaagata gcgagattga tgcacacaaa caggagtctg 3601 gtatcgctcc taactttgta cacagccaag acggtagcca ccttcgtaag actgtagtgt 3661 gggcacacga gaagtacgga atcgaatctt ttgcactgat tcacgactcc ttcggtacca 3721 ttccggctga cgctgcgaac ctgttcaaag cagtgcgcga aactatggtt gacacatatg 3781 agtcttgtga tgtactggct gatttctacg accagttcgc tgaccagttg cacgagtctc 3841 aattggacaa aatgccagca cttccggcta aaggtaactt gaacctccgt gacatcttag 3901 agtcggactt cgcgttcgcg taacgccaaa tcaatacgac tccggatcct gcccatttgg 3961 cttttccctt gtctcgtgtc ttttccgtgg aaaggttccc ggagtaatct gatggcacag 4021 cagggaggtg cgcctgcagg ttggttagga aggggggatg atgtaaaaga agaaaatggg 4081 gggattcgag cccgggcaca gcaaggtctt ctgaaattca tgtttttttt ttttttactc 4141 tgcattgcag tctccgctct tatttagttt tgctttacgt aaggtctcgt tgctgccata 4201 aaataagctt agaggagaaa gaatagtaac cctttcatca aagaaaatag ttcaaacgaa 4261 tatgcgcgaa atcgtctgcg ttcaggctgg ccaatgcggt aaccagatcg gctcaaagtt 4321 ctgggaggtg atccggccaa gcttggatgg attgcacgca ggttctccgg ccgcttgggt 4381 ggagaggcta ttcggctatg actgggcaca acagacaatc ggctgctctg atgccgccgt 4441 gttccggctg tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc 4501 cctgaatgaa ctgcaggacg aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc 4561 ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg gactggctgc tattgggcga 4621 agtgccgggg caggatctcc tgtcatctca ccttgctcct gccgagaaag tatccatcat 4681 ggctgatgca atgcggcggc tgcatacgct tgatccggct acctgcccat tcgaccacca 4741 agcgaaacat cgcatcgagc gagcacgtac tcggatggaa gccggtcttg tcgatcagga 4801 tgatctggac gaagagcatc aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc 4861 gcgcatgccc gacggcgagg atctcgtcgt gacccatggc gatgcctgct tgccgaatat 4921 catggtggaa aatggccgct tttctggatt catcgactgt ggccggctgg gtgtggcgga 4981 ccgctatcag gacatagcgt tggctacccg tgatattgct gaagagcttg gcggcgaatg 5041 ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc gattcgcagc gcatcgcctt 5101 ctatcgcctt cttgacgagt tcttctgagc gggactctgg ggttcggatc ctaacaccgg 5161 gttgtgtggc caaaattgtt ctgtagtcgc tgtgagttga cacggctagt gcttatgatt 5221 ttcctcgcgt gtggtgcctg tactcagccc tatgccttat ttgcaacaca tttacgtaca 5281 gcgcacaaga gaagagaaga tcacttgaag ataataaata tagggttgta ggcatcttgt 5341 ttaactcaaa ttttctcgtc ttggtgtgtc gacatgattg aaatagtgcc accagttgtg 5401 tttgatgcgt ttgttatcta tgcagtattc tgcaaggggg cacagcaagg tcttctgaaa 5461 ttcatgtttt tttttttttt actctgcatt gcagtctccg ctcttattta gttttgcttt 5521 acgtaaggtc tcgttgctgc cataaaataa gcctagaact agagctttca tatgtctaga 5581 atggactata aggaccacga cggagactac aaggatcatg atattgatta caaagacgat 5641 gacgataaga tggccccaaa gaagaagcgg aaggtcggta tccacggagt cccagcagcc 5701 gacaagaagt acagcatcgg cctggacatc ggcaccaact ctgtgggctg ggccgtgatc 5761 accgacgagt acaaggtgcc cagcaagaaa ttcaaggtgc tgggcaacac cgaccggcac 5821 agcatcaaga agaacctgat cggagccctg ctgttcgaca gcggcgaaac agccgaggcc 5881 acccggctga agagaaccgc cagaagaaga tacaccagac ggaagaaccg gatctgctat 5941 ctgcaagaga tcttcagcaa cgagatggcc aaggtggacg acagcttctt ccacagactg 6001 gaagagtcct tcctggtgga agaggataag aagcacgagc ggcaccccat cttcggcaac 6061 atcgtggacg aggtggccta ccacgagaag taccccacca tctaccacct gagaaagaaa 6121 ctggtggaca gcaccgacaa ggccgacctg cggctgatct atctggccct ggcccacatg 6181 atcaagttcc ggggccactt cctgatcgag ggcgacctga accccgacaa cagcgacgtg 6241 gacaagctgt tcatccagct ggtgcagacc tacaaccagc tgttcgagga aaaccccatc 6301 aacgccagcg gcgtggacgc caaggccatc ctgtctgcca gactgagcaa gagcagacgg 6361 ctggaaaatc tgatcgccca gctgcccggc gagaagaaga atggcctgtt cggaaacctg 6421 attgccctga gcctgggcct gacccccaac ttcaagagca acttcgacct ggccgaggat 6481 gccaaactgc agctgagcaa ggacacctac gacgacgacc tggacaacct gctggcccag 6541 atcggcgacc agtacgccga cctgtttctg gccgccaaga acctgtccga cgccatcctg 6601 ctgagcgaca tcctgagagt gaacaccgag atcaccaagg cccccctgag cgcctctatg 6661 atcaagagat acgacgagca ccaccaggac ctgaccctgc tgaaagctct cgtgcggcag 6721 cagctgcctg agaagtacaa agagattttc ttcgaccaga gcaagaacgg ctacgccggc 6781 tacattgacg gcggagccag ccaggaagag ttctacaagt tcatcaagcc catcctggaa 6841 aagatggacg gcaccgagga actgctcgtg aagctgaaca gagaggacct gctgcggaag 6901 cagcggacct tcgacaacgg cagcatcccc caccagatcc acctgggaga gctgcacgcc 6961 attctgcggc ggcaggaaga tttttaccca ttcctgaagg acaaccggga aaagatcgag 7021 aagatcctga ccttccgcat cccctactac gtgggccctc tggccagggg aaacagcaga 7081 ttcgcctgga tgaccagaaa gagcgaggaa accatcaccc cctggaactt cgaggaagtg 7141 gtggacaagg gcgcttccgc ccagagcttc atcgagcgga tgaccaactt cgataagaac 7201 ctgcccaacg agaaggtgct gcccaagcac agcctgctgt acgagtactt caccgtgtat 7261 aacgagctga ccaaagtgaa atacgtgacc gagggaatga gaaagcccgc cttcctgagc 7321 ggcgagcaga aaaaggccat cgtggacctg ctgttcaaga ccaaccggaa agtgaccgtg 7381 aagcagctga aagaggacta cttcaagaaa atcgagtgct tcgactccgt ggaaatctcc 7441 ggcgtggaag atcggttcaa cgcctccctg ggcacatacc acgatctgct gaaaattatc 7501 aaggacaagg acttcctgga caatgaggaa aacgaggaca ttctggaaga tatcgtgctg 7561 accctgacac tgtttgagga cagagagatg atcgaggaac ggctgaaaac ctatgcccac 7621 ctgttcgacg acaaagtgat gaagcagctg aagcggcgga gatacaccgg ctggggcagg 7681 ctgagccgga agctgatcaa cggcatccgg gacaagcagt ccggcaagac aatcctggat 7741 ttcctgaagt ccgacggctt cgccaacaga aacttcatgc agctgatcca cgacgacagc 7801 ctgaccttta aagaggacat ccagaaagcc caggtgtccg gccagggcga tagcctgcac 7861 gagcacattg ccaatctggc cggcagcccc gccattaaga agggcatcct gcagacagtg 7921 aaggtggtgg acgagctcgt gaaagtgatg ggccggcaca agcccgagaa catcgtgatc 7981 gaaatggcca gagagaacca gaccacccag aagggacaga agaacagccg cgagagaatg 8041 aagcggatcg aagagggcat caaagagctg ggcagccaga tcctgaaaga acaccccgtg 8101 gaaaacaccc agctgcagaa cgagaagctg tacctgtact acctgcagaa tgggcgggat 8161 atgtacgtgg accaggaact ggacatcaac cggctgtccg actacgatgt ggaccatatc 8221 gtgcctcaga gctttctgaa ggacgactcc atcgacaaca aggtgctgac cagaagcgac 8281 aagaaccggg gcaagagcga caacgtgccc tccgaagagg tcgtgaagaa gatgaagaac 8341 tactggcggc agctgctgaa cgccaagctg attacccaga gaaagttcga caatctgacc 8401 aaggccgaga gaggcggcct gagcgaactg gataaggccg gcttcatcaa gagacagctg 8461 gtggaaaccc ggcagatcac aaagcacgtg gcacagatcc tggactcccg gatgaacact 8521 aagtacgacg agaatgacaa gctgatccgg gaagtgaaag tgatcaccct gaagtccaag 8581 ctggtgtccg atttccggaa ggatttccag ttttacaaag tgcgcgagat caacaactac 8641 caccacgccc acgacgccta cctgaacgcc gtcgtgggaa ccgccctgat caaaaagtac 8701 cctaagctgg aaagcgagtt cgtgtacggc gactacaagg tgtacgacgt gcggaagatg 8761 atcgccaaga gcgagcagga aatcggcaag gctaccgcca agtacttctt ctacagcaac 8821 atcatgaact ttttcaagac cgagattacc ctggccaacg gcgagatccg gaagcggcct 8881 ctgatcgaga caaacggcga aaccggggag atcgtgtggg ataagggccg ggattttgcc 8941 accgtgcgga aagtgctgag catgccccaa gtgaatatcg tgaaaaagac cgaggtgcag 9001 acaggcggct tcagcaaaga gtctatcctg cccaagagga acagcgataa gctgatcgcc 9061 agaaagaagg actgggaccc taagaagtac ggcggcttcg acagccccac cgtggcctat 9121 tctgtgctgg tggtggccaa agtggaaaag ggcaagtcca agaaactgaa gagtgtgaaa 9181 gagctgctgg ggatcaccat catggaaaga agcagcttcg agaagaatcc catcgacttt 9241 ctggaagcca agggctacaa agaagtgaaa aaggacctga tcatcaagct gcctaagtac 9301 tccctgttcg agctggaaaa cggccggaag agaatgctgg cctctgccgg cgaactgcag 9361 aagggaaacg aactggccct gccctccaaa tatgtgaact tcctgtacct ggccagccac 9421 tatgagaagc tgaagggctc ccccgaggat aatgagcaga aacagctgtt tgtggaacag 9481 cacaagcact acctggacga gatcatcgag cagatcagcg agttctccaa gagagtgatc 9541 ctggccgacg ctaatctgga caaagtgctg tccgcctaca acaagcaccg ggataagccc 9601 atcagagagc aggccgagaa tatcatccac ctgtttaccc tgaccaatct gggagcccct 9661 gccgccttca agtactttga caccaccatc gaccggaaga ggtacaccag caccaaagag 9721 gtgctggacg ccaccctgat ccaccagagc atcaccggcc tgtacgagac acggatcgac 9781 ctgtctcagc tgggaggcga caaaaggccg gcggccacga aaaaggccgg ccaggcaaaa 9841 aagaaaaagt aactcgagag gtgcagagcc agccttctta ttcggccttg aattgatcat 9901 atgcggatta gaaaaacaac ttaaatgtga aagtgggtct taaggatccc aaaattgttc 9961 tgtagtcgct gtgagttgac acggctagtg cttatgattt tcctcgcgtg tggtgcctgt 10021 actcagccct atgccttatt tgcaacacat ttacgtacag cgcacaagag aagagaagat 10081 cacttgaaga taataaatat agggttgtag gcatcttgtt taactcaaat tttctcgtct 10141 tggtgtgtcg acatgattga aatagtgcca ccagttgtgt ttgatgcgtt tgttatctat 10201 gcagtattct gcaaggcctt gcaggcatgc aagctagctt gtattctata gtgtcaccta 10261 aatcgtatgt gtatgataca taaggttatg tattaattgt agccgcgttc taacgacaat 10321 atgtacaagc ctaattgtgt agcatctggc ttactgaagc agaccctatc atctctctcg 10381 taaactgccg tcagagtcgg tttggttgga cgaaccttct gagtttctgg taacgccgtt 10441 ccgcaccccg gaaatggtca gcgaaccaat cagcagggtc atcgctagcc agatcctcta 10501 cgccggacgc atcgtggccg gcatcaccgg cgccacaggt gcggttgctg gcgcctatat 10561 cgccgacatc accgatgggg aagatcgggc tcgccacttc gggctcatga gcgcttgttt 10621 cggcgtgggt atggtggcag gccccgtggc cgggggactg ttgggcgcca tctccttgca 10681 ccattccttg cggcggcggt gctcaacggc ctcaacctac tactgggctg cttcctaatg 10741 caggagtcgc ataagggaga gcgtcgatat ggtgcactct cagtacaatc tgctctgatg 10801 ccgcatagtt aagccagccc cgacacccgc caacacccgc tgacgcgccc tgacgggctt 10861 gtctgctccc ggcatccgct tacagacaag ctgtgaccgt ctccgggagc tgcatgtgtc 10921 agaggttttc accgtcatca ccgaaacgcg cgagacgaaa gggcctcgtg atacgcctat 10981 ttttataggt taatgtcatg ataataatgg tttcttagac gtcaggtggc acttttcggg 11041 gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc 11101 tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag agtatgagta 11161 ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg 11221 ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg 11281 gttacatcga actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac 11341 gttttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtattg 11401 acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt 11461 actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg 11521 ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg atcggaggac 11581 cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt 11641 gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgtag 11701 caatggcaac aacgttgcgc aaactattaa ctggcgaact acttactcta gcttcccggc 11761 aacaattaat agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc 11821 ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggg tctcgcggta 11881 tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg 11941 ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga 12001 ttaagcattg gtaactgtca gaccaagttt actcatatat actttagatt gatttaaaac 12061 ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa 12121 tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat 12181 cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc 12241 taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg 12301 gcttcagcag agcgcagata ccaaatactg tccttctagt gtagccgtag ttaggccacc 12361 acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg 12421 ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg 12481 ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa 12541 cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg 12601 aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga 12661 gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct 12721 gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca 12781 gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgttctttc 12841 ctgcgttatc ccctgattct gtggataacc gtattaccgc ctttgagtga gctgataccg 12901 ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg gaagagcgcc 12961 caatacgcaa accgcctctc cccgcgcgtt ggccgattca ttaatgcag // LOCUS pPOTcruzi v1 blast-blast mNeonGreen 5949 bp ds-DNA circular SYN 24-JUL-2017 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS . SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 5949) AUTHORS . TITLE Direct Submission JOURNAL Exported Jan 18, 2018 from SnapGene Viewer 4.0.6 http://www.snapgene.com COMMENT C terminal tag: For: ggttctggtagtggttcc Rev: ccaatttgagagacctgtgc N terminal tag For: gtataatgcagacctgctgc Rev: actacccgatcctgatcc FEATURES Location/Qualifiers source 1..5949 /organism="synthetic DNA construct" /mol_type="other DNA" primer_bind 16..35 /label=Up Fw 5'UTR 36..410 /locus_tag="H2B intergenic" CDS join(417..688,690..727,729..806) /locus_tag="BLAST" /label=BLAST 5'UTR 816..1150 /locus_tag="histon h4 IGR" misc_feature 1161..1190 /locus_tag="GS linker(1)" primer_bind 1161..1180 /label=Down Fw CDS 1197..1901 /locus_tag="mNG" /label=mNG misc_feature 1908..1937 /locus_tag="GS linker" primer_bind complement(1918..1937) /label=Up Rev 5'UTR 1947..2281 /locus_tag="histon h4 IGR(1)" /label=Histone H4 intergenic region CDS join(2289..2560,2562..2599,2601..2678) /locus_tag="BLAST(1)" /label=BLAST(1) 5'UTR 2688..3036 /locus_tag="H2A IGR" /label=5' UTR - histone H2A intergenic region primer_bind complement(3037..3056) /locus_tag="#162" primer_bind complement(3037..3056) /label=Down Rev primer_bind complement(3075..3100) /locus_tag="SP6" rep_origin complement(3494..4176) /direction=LEFT /locus_tag="ColE1 origin" CDS complement(4274..4933) /locus_tag="AmpR" /label=AmpR rep_origin complement(5446..5752) /direction=LEFT /locus_tag="F1 ori" CDS complement(5770..5838) /locus_tag="LacZ alpha" /label=LacZ alpha primer_bind 5909..5926 /locus_tag="M13-fwd" primer_bind 5925..3 /locus_tag="T7" ORIGIN 1 gggcgaattg ggcccgtata atgcagacct gctgcatcct ttgtggcgtt tcttgttttt 61 acaagcgtct agaagcactt tttttcgttt tctttttttt gatttaggct ccttcaccct 121 cttccgtttt tttttttttt taatgccgcg ggacgtagcc ctctgggcgg ctcgtggtgc 181 gcgtgtggtt ttcatgacgg gccgcacgtg aagtatagag ggagtctgat tgctgtgcgg 241 gtggaggtgc ccgatgctct cgccctgttt gtgtgtctgt gtgtttggct gcagctgctc 301 ttgccggctc tccgctgatg tgctcttttt cctctcattt tctcctgcgc ttttcgactg 361 ctcgcgcaag atctctgcga acaagcaacc agaaccaagt acgaaccaag gaattcatgc 421 ctttgtctca agaagaatcc accctcattg aaagagcaac ggctacaatc aacagcatcc 481 ccatctctga agactacagc gtcgccagcg cagctctctc tagcgacggc cgcatcttca 541 ctggtgtcaa tgtatatcat tttactgggg gaccttgtgc agaactcgtg gtgctgggca 601 ctgctgctgc tgcggcagct ggcaacctga cttgtatcgt cgcgatcgga aatgagaaca 661 ggggcatctt gagcccctgc ggacggtgcc gacaggtgct tctcgatctg catcctggga 721 tcaaagccat agtgaaggac agtgatggac agccgacggc agttgggatt cgtgaattgc 781 tgccctctgg ttatgtgtgg gagggctaac catggggacc cacttgttag aggcatcagt 841 cgcggcgggt ggactggcgg caacgccatg cccccgcaac tatagataga ttagcaacgg 901 tgataggaag ggagcaagac acttggggct gctcggctga cggccacacg tagatatata 961 tatatatata cacatgtgtg tgtgtgcgtg tgcagtgagg cacgctcccc tctgtgatca 1021 ccagtcctca tgggcgtcca gctgcacctt ctctctcttt ctctcctttt tttcctttct 1081 ttttgcccct cgctctcaca caccgcgcag aaccaagcaa ccaagcacac agctaaacaa 1141 acacagcaag ggtacctatg ggttctggta gtggttccgg ttccggttct aagcttgtct 1201 cgaaaggtga ggaagacaat atggccagct tgccagcaac gcacgagctt catatatttg 1261 ggtcgatcaa cggtgtggat ttcgacatgg tgggtcaagg cacaggtaat cctaatgacg 1321 ggtatgaaga actcaatttg aagagcacta agggagatct ccagttttcc ccgtggattc 1381 ttgtgccgca tattggttat ggttttcatc agtacttacc ataccccgat ggaatgtcgc 1441 ccttccaagc agctatggtg gacggctcgg ggtatcaagt acatcgcacc atgcaatttg 1501 aggacggagc atccctcacc gtaaactacc gctatacgta tgaggggagc cacataaaag 1561 gtgaggctca agttaaaggc acaggcttcc cagctgacgg cccggtcatg accaattcat 1621 taaccgctgc tgattggtgc cgttctaaga agacctaccc taacgacaag acgatcatta 1681 gtacttttaa gtggagttat accacaggaa acggaaagcg gtatcgaagc actgctcgaa 1741 caacctacac atttgcgaag ccaatggccg cgaactactt gaagaaccag ccaatgtatg 1801 tgtttcgaaa aacggaactg aagcattcga aaaccgagct taattttaaa gagtggcaga 1861 aggcgtttac agacgtgatg gggatggacg aattgtataa gggatccgga tcaggatctg 1921 gatcaggatc gggtagttaa gagctcggac ccacttgtta gaggcatcag tcgcggcggg 1981 tggactggcg gcaacgccat gcccccgcaa ctatagatag attagcaacg gtgataggaa 2041 gggagcaaga cacttggggc tgctcggctg acggccacac gtagatatat atatatatat 2101 acacatgtgt gtgtgtgcgt gtgcagtgag gcacgctccc ctctgtgatc accagtcctc 2161 atgggcgtcc agctgcacct tctctctctt tctctccttt ttttcctttc tttttgcccc 2221 tcgctctcac acaccgcgca gaaccaagca accaagcaca cagctaaaca aacacagcaa 2281 ggctagccat gcctttgtct caagaagaat ccaccctcat tgaaagagca acggctacaa 2341 tcaacagcat ccccatctct gaagactaca gcgtcgccag cgcagctctc tctagcgacg 2401 gccgcatctt cactggtgtc aatgtatatc attttactgg gggaccttgt gcagaactcg 2461 tggtgctggg cactgctgct gctgcggcag ctggcaacct gacttgtatc gtcgcgatcg 2521 gaaatgagaa caggggcatc ttgagcccct gcggacggtg ccgacaggtg cttctcgatc 2581 tgcatcctgg gatcaaagcc atagtgaagg acagtgatgg acagccgacg gcagttggga 2641 ttcgtgaatt gctgccctct ggttatgtgt gggagggcta actcgagagc ggccttcagg 2701 ccaggcgcag ggagggggcg caacatgtta gatgcacctg ggtggccgtc gtcgctgagc 2761 cacggtgacc cacacgtagc cctatagaat aggattggtt tgttgttttc ttttcgtttt 2821 ctactgcctc tctcccaact gttgtgtagg tctctgcggc gcctcggcat cggtgcttgg 2881 ccgcattgtg gcgtgccgtg cgctttttcc tctgctttgg ccgcaggtct cctgcccccg 2941 cccttctctc ctccctttgt ttcccaagat caaagccaag ccaaggaacc caaccatact 3001 ttatttgtat tttttttttg tttttcgcag acaagggcac aggtctctca aattggatgc 3061 atagcttgag tattctatag tgtcacctaa atagcttggc gtaatcatgg tcatagctgt 3121 ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa 3181 agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac 3241 tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg 3301 cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc 3361 gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat 3421 ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca 3481 ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc 3541 atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc 3601 aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg 3661 gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta 3721 ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg 3781 ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac 3841 acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag 3901 gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat 3961 ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat 4021 ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc 4081 gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt 4141 ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct 4201 agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt 4261 ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc 4321 gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac 4381 catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat 4441 cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg 4501 cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata 4561 gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta 4621 tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt 4681 gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag 4741 tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa 4801 gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc 4861 gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt 4921 taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc 4981 tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta 5041 ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa 5101 taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca 5161 tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac 5221 aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgatgcggtg tgaaataccg 5281 cacagatgcg taaggagaaa ataccgcatc aggaaattgt aagcgttaat attttgttaa 5341 aattcgcgtt aaatttttgt taaatcagct cattttttaa ccaataggcc gaaatcggca 5401 aaatccctta taaatcaaaa gaatagaccg agatagggtt gagtgttgtt ccagtttgga 5461 acaagagtcc actattaaag aacgtggact ccaacgtcaa agggcgaaaa accgtctatc 5521 agggcgatgg cccactacgt gaaccatcac cctaatcaag ttttttgggg tcgaggtgcc 5581 gtaaagcact aaatcggaac cctaaaggga gcccccgatt tagagcttga cggggaaagc 5641 cggcgaacgt ggcgagaaag gaagggaaga aagcgaaagg agcgggcgct agggcgctgg 5701 caagtgtagc ggtcacgctg cgcgtaacca ccacacccgc cgcgcttaat gcgccgctac 5761 agggcgcgtc cattcgccat tcaggctgcg caactgttgg gaagggcgat cggtgcgggc 5821 ctcttcgcta ttacgccagc tggcgaaagg gggatgtgct gcaaggcgat taagttgggt 5881 aacgccaggg ttttcccagt cacgacgttg taaaacgacg gccagtgaat tgtaatacga 5941 ctcactata // LOCUS pPOTv7 blast-blast mScarlet 5516 bp ds-DNA circular SYN 13-JUL-2017 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS . SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 5516) AUTHORS . TITLE Direct Submission JOURNAL Exported Jan 18, 2018 from SnapGene Viewer 4.0.6 http://www.snapgene.com COMMENT C terminal tag: For: ggttctggtagtggttcc Rev: ccaatttgagagacctgtgc N terminal tag For: gtataatgcagacctgctgc Rev: actacccgatcctgatcc FEATURES Location/Qualifiers source 1..5516 /organism="synthetic DNA construct" /mol_type="other DNA" primer_bind 16..35 /label=#161 5'UTR 36..164 /label=Actin 5' CDS join(171..442,444..481,483..560) /label=BLAST 3'UTR 570..718 /label=Ald3'UTR (short) 5'UTR join(725..744,746..913) /label=Aldolase 5' CDS 921..959 /codon_start=1 /label=Feature 22 /translation="MGSGSGSGSGSKL" misc_feature 924..953 /label=GS linker(1) CDS 960..1652 /codon_start=1 /label=trypmScarlet /translation="VSKGEAVIKEFMRFKVHMEGSMNGHEFEIEGEGEGRPYEGTQTAK LKVTKGGPLPFSWDILSPQFMYGSRAFTKHPADIPDYYKQSFPEGFKWERVMNFEDGGA VTVTQDTSLEDGTLIYKVKLRGTNFPPDGPVMQKKTMGWEASTERLYPEDGVLKGDIKM ALRLKDGGRYLADFKTTYKAKKPVQMPGAYNVDRKLDITSHNEDYTVVEQYERSEGRHS TGGMDELYK" misc_feature 1659..1688 /label=GS linker 3'UTR join(1698..1709,1711..1893,1895..2048) /label=PFR2 IGS CDS join(2056..2327,2329..2366,2368..2445) /label=BLAST(1) 3'UTR 2455..2603 /label=Ald3'UTR (short)(1) primer_bind complement(2604..2623) /label=#162 primer_bind complement(2642..2667) /label=SP6 primer_bind complement(2673..2693) /label=M13-rev misc_binding complement(2699..2721) /label=LacO rep_origin complement(3061..3743) /direction=LEFT /label=ColE1 origin CDS complement(3841..4500) /label=AmpR rep_origin complement(5013..5319) /direction=LEFT /label=F1 ori CDS complement(5337..5405) /label=LacZ alpha primer_bind 5476..5493 /label=M13-fwd primer_bind 5492..3 /label=T7 ORIGIN 1 gggcgaattg ggcccgtata atgcagacct gctgcaagct aggggcacag caaggtcttc 61 tgaaattcat gttttttttt tttttactct gcattgcagt ctccgctctt atttagtttt 121 gctttacgta aggtctcgtt gctgccataa aataagctct agtggaattc atgcctttgt 181 ctcaagaaga atccaccctc attgaaagag caacggctac aatcaacagc atccccatct 241 ctgaagacta cagcgtcgcc agcgcagctc tctctagcga cggccgcatc ttcactggtg 301 tcaatgtata tcattttact gggggacctt gtgcagaact cgtggtgctg ggcactgctg 361 ctgctgcggc agctggcaac ctgacttgta tcgtcgcgat cggaaatgag aacaggggca 421 tcttgagccc ctgcggacgg tgccgacagg tgcttctcga tctgcatcct gggatcaaag 481 ccatagtgaa ggacagtgat ggacagccga cggcagttgg gattcgtgaa ttgctgccct 541 ctggttatgt gtgggagggc taaccatggt gcccatttag ttggcttttc ccttgtctcg 601 tgtcttttcc gtggaaaggt tcccggagta atctgatggc acagcaggga ggtgcgcctg 661 caggttggtt aggaaggggg gatgatgtaa aagaagaaaa tggggggatt cgagccccac 721 gcgttgctca agctgtgtag cgcaggcgtt tccttacata tttctctaac aggcacggaa 781 gcctaacaaa tacacttggc ttattttttt gccccctcat gtcttgtaca aatatttgcg 841 atagcttagc tatcagccac attaatcaaa caagtatacc aacaagcccg aaaacagaaa 901 ctcaactgca acgggtacct atgggttctg gtagtggttc cggttccggt tctaagcttg 961 taagcaaagg ggaggctgtc ataaaggaat ttatgcgatt taaagtgcac atggaaggga 1021 gtatgaacgg ccatgagttt gaaatagaag gagagggaga ggggcgccct tacgaaggta 1081 ctcagacggc gaagctcaaa gtgactaagg gaggtcctct gccgtttagc tgggatatac 1141 tttcacccca gttcatgtat ggcagccgcg cgttcacgaa gcaccccgca gatatccccg 1201 attactataa acagtcattt cctgaggggt tcaagtggga acgagtaatg aactttgagg 1261 atggtggcgc tgtaacggta acacaagata cctcgctcga agacggtacg ctcatttaca 1321 aagtgaagct gcgcggaaca aatttccctc ctgatggacc agtaatgcag aaaaagacta 1381 tgggatggga agcctcgacg gaacgtctct acccggagga cggggtgctg aaaggggata 1441 ttaaaatggc actccgactg aaagacggcg gcaggtatct tgctgatttt aaaaccacgt 1501 acaaagctaa aaagccagtg cagatgcctg gagcgtataa cgtagaccga aaactcgaca 1561 ttacgtcgca taacgaggac tatacggtag tcgaacagta tgagaggtct gaagggaggc 1621 attcaactgg tggtatggat gagttgtata agggatccgg atcaggatct ggatcaggat 1681 cgggtagtta agagctccgc tgcgcttaaa tgtctttcat tataatcaat gtataacctt 1741 tatgtagtat ttcaatctat gccgctgtgt acgtgcactg cggtgcctat ccttcggcat 1801 tagagagtca ctgtttgtgt agatcgtagc tgcatgtctg acatctggtt cattagtgcc 1861 gttttgagtc tgcactgttg tgcaacttga tattcatcac tgcacaccaa ccgtgtgttt 1921 cctcctttgg gtgcaccctt taatccttgt cttctccttt tttgtctctt tccccctcga 1981 aaaggtgtca aactactgcc gcataaacta cggttatccc aaatttaaga gaaagcaata 2041 aagcatcagc tagcnatgcc tttgtctcaa gaagaatcca ccctcattga aagagcaacg 2101 gctacaatca acagcatccc catctctgaa gactacagcg tcgccagcgc agctctctct 2161 agcgacggcc gcatcttcac tggtgtcaat gtatatcatt ttactggggg accttgtgca 2221 gaactcgtgg tgctgggcac tgctgctgct gcggcagctg gcaacctgac ttgtatcgtc 2281 gcgatcggaa atgagaacag gggcatcttg agcccctgcg gacggtgccg acaggtgctt 2341 ctcgatctgc atcctgggat caaagccata gtgaaggaca gtgatggaca gccgacggca 2401 gttgggattc gtgaattgct gccctctggt tatgtgtggg agggctaact cgagtgccca 2461 tttagttggc ttttcccttg tctcgtgtct tttccgtgga aaggttcccg gagtaatctg 2521 atggcacagc agggaggtgc gcctgcaggt tggttaggaa ggggggatga tgtaaaagaa 2581 gaaaatgggg ggattcgagc cccgcacagg tctctcaaat tggatgcata gcttgagtat 2641 tctatagtgt cacctaaata gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa 2701 ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg 2761 gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca 2821 gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg 2881 tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg 2941 gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg 3001 ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa 3061 ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg 3121 acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc 3181 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc 3241 ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc 3301 ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg 3361 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc 3421 actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga 3481 gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc 3541 tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac 3601 caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg 3661 atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc 3721 acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa 3781 ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta 3841 ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt 3901 tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag 3961 tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag caataaacca 4021 gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc 4081 tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt 4141 tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag 4201 ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt 4261 tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat 4321 ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt 4381 gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc 4441 ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat 4501 cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag 4561 ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt 4621 ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg 4681 gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta 4741 ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc 4801 gcgcacattt ccccgaaaag tgccacctga tgcggtgtga aataccgcac agatgcgtaa 4861 ggagaaaata ccgcatcagg aaattgtaag cgttaatatt ttgttaaaat tcgcgttaaa 4921 tttttgttaa atcagctcat tttttaacca ataggccgaa atcggcaaaa tcccttataa 4981 atcaaaagaa tagaccgaga tagggttgag tgttgttcca gtttggaaca agagtccact 5041 attaaagaac gtggactcca acgtcaaagg gcgaaaaacc gtctatcagg gcgatggccc 5101 actacgtgaa ccatcaccct aatcaagttt tttggggtcg aggtgccgta aagcactaaa 5161 tcggaaccct aaagggagcc cccgatttag agcttgacgg ggaaagccgg cgaacgtggc 5221 gagaaaggaa gggaagaaag cgaaaggagc gggcgctagg gcgctggcaa gtgtagcggt 5281 cacgctgcgc gtaaccacca cacccgccgc gcttaatgcg ccgctacagg gcgcgtccat 5341 tcgccattca ggctgcgcaa ctgttgggaa gggcgatcgg tgcgggcctc ttcgctatta 5401 cgccagctgg cgaaaggggg atgtgctgca aggcgattaa gttgggtaac gccagggttt 5461 tcccagtcac gacgttgtaa aacgacggcc agtgaattgt aatacgactc actata // LOCUS pTbBLAST 4539 bp ds-DNA circular SYN 18-JAN-2018 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS pTbBLAST SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 4539) AUTHORS MCT TITLE Direct Submission JOURNAL Exported Jan 18, 2018 from SnapGene Viewer 4.0.6 http://www.snapgene.com FEATURES Location/Qualifiers source 1..4539 /organism="synthetic DNA construct" /mol_type="other DNA" promoter 305..323 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" misc_feature 411..1040 /label=Tb-alpha-beta-tubulin IR CDS 1048..1446 /codon_start=1 /gene="Aspergillus terreus BSD" /product="blasticidin S deaminase" /label=BSD /note="confers resistance to blasticidin" /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI KAIVKDSDGQPTAVGIRELLPSGYVWEG" misc_feature 1460..1901 /label=TbActin IR protein_bind 2347..2363 /label=lac operator /bound_moiety="lac repressor encoded by lacI" /note="The lac repressor binds to the lac operator to inhibit transcription in E. coli. This inhibition can be relieved by adding lactose or isopropyl-beta-D-thiogalactopyranoside (IPTG)." promoter complement(2371..2401) /label=lac UV5 promoter /note="E. coli lac promoter with an ""up"" mutation" protein_bind 2416..2437 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." rep_origin complement(2728..3316) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" CDS complement(3487..4347) /codon_start=1 /gene="bla" /product="beta-lactamase" /label=AmpR /note="confers resistance to ampicillin, carbenicillin, and related antibiotics" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS LIKHW" promoter complement(4348..4452) /gene="bla" /label=AmpR promoter ORIGIN 1 gcccctgcag ccgaattata ttatttttgc caaataattt ttaacaaaag ctctgaagtc 61 ttcttcattt aaattcttag atgatacttc atctggaaaa ttgtcccaat tagtagcatc 121 acgctgtgag taagttctaa accatttttt tattgttgta ttatctctaa tcttactact 181 cgatgagttt tcggtattat ctctattttt aacttggagc aggttccatt cattgttttt 241 ttcatcatag tgaataaaat caactgcttt aacacttgtg cctgaacacc atatccatcc 301 ggcgtaatac gactcactat agggagagcg gccgccagat cttccggatg gctcgagttt 361 ttcagcaaga taaatgtata atgcagacct gctgccctag gtgaggatcc acgcggacgg 421 ggcatttccc gttcgtcatt agcagtaggt aatgaagatg tttgtttctc gtcccctttc 481 tccttcgtcc ttctgtcatt ttgttctttt gtgtttatgt tttgttgttg ttttctttaa 541 tttttttttt cttccacgtt tgtgtacatc cgcgcgccac tctattcaga gagccacgga 601 tagtagagga ggtgggaagg gtatatgagg gacacgcgta ccatgatgtg ggatgtattg 661 gggtccctgt ctgtccttac gtgactatgt atgaaccgtc acgtgtaaga tgagctagtg 721 agatcaacag tacaactcat caacacgcct tcttctcgtt aaatgtacac aatcttgatc 781 ctccaccttt atgggtccca ttgtttgcct cttccgctgt gtggagtgcg cctacacgca 841 cttctcactt cgtaagtggt ggtggcgtaa gtattgccta atgttgactc tatattctcc 901 tctcctcacc ccctcgcggt gctgatttct gacagatctt caaacactag tttaagcaaa 961 ggactattca tccgtttata ttagcaacag taggtactag caccactaac aacaacaaca 1021 aagcacttct atttatttat cggtaccatg gccaagcctt tgtctcaaga agaatccacc 1081 ctcattgaaa gagcaacggc tacaatcaac agcatcccca tctctgaaga ctacagcgtc 1141 gccagcgcag ctctctctag cgacggccgc atcttcactg gtgtcaatgt atatcatttt 1201 actgggggac cttgtgcaga actcgtggtg ctgggcactg ctgctgctgc ggcagctggc 1261 aacctgactt gtatcgtcgc gatcggaaat gagaacaggg gcatcttgag cccctgcgga 1321 cggtgccgac aggtgcttct cgatctgcat cctgggatca aagccatagt gaaggacagt 1381 gatggacagc cgacggcagt tgggattcgt gaattgctgc cctctggtta tgtgtgggag 1441 ggctaagaat tcgctagcga tccctcggag atcctaacac cgggttgtgt ggccaaaatt 1501 gttctgtagt cgctgtgagt tgacacggct agtgcttatg attttcctcg cgtgtggtgc 1561 ctgtactcag ccctatgcct tatttgcaac acatttacgt acagcgcaca agagaagaga 1621 agatcacttg aagataataa atatagggtt gtaggcatct tgtttaactc aaattttctc 1681 gtcttggtgt gtcgacatga ttgaaatagt gccaccagtt gtgtttgatg cgtttgttat 1741 ctatgcagta ttcctagcca gatcctctac gccggacgca tcgtggccgg catcaccggc 1801 gccacaggtg cggttgctgg cgcctatatc gccgacatca ccgatgggga agatcgggct 1861 cgccacttcg ggctcatgag cgcttgtttc ggcgtgggta tggtggcagg ccgcacaggt 1921 ctctcaaatt ggatttatct ttctagaaga tctcctacaa tattctcagc tgccatggaa 1981 aatcgatgtt cttcttttat tctctcaaga ttttcaggct gtatattaaa acttatatta 2041 agaactatgc taaccacctc atcaggaacc gttgtaggtg gcgtgggttt tcttggcaat 2101 cgactctcat gaaaactacg agctaaatat tcaatatgtt cctcttgacc aactttattc 2161 tgcatttttt ttgaacgagg tttagagcaa gcttcaggaa actgagacag gaattttatt 2221 aaaaatttaa attttgaaga aagttcaggg ttaatagcat ccattttttg ctttgcaagt 2281 tcctcagcat tcttaacaaa agacgtctct tttgacatgt ttaaagttta aacctcctgt 2341 gtgaaattgt tatccgctca caattccaca cattatacga gccggaagca taaagtgtaa 2401 agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct cactgccaat 2461 tgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg 2521 gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc 2581 ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac 2641 agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa 2701 ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca 2761 caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc 2821 gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata 2881 cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta 2941 tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca 3001 gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga 3061 cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg 3121 tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagga cagtatttgg 3181 tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg 3241 caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag 3301 aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa 3361 cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat 3421 ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc 3481 tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc 3541 atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc 3601 tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc 3661 aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc 3721 catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt 3781 gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc 3841 ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa 3901 aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt 3961 atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg 4021 cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc 4081 gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa 4141 agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt 4201 gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt 4261 caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag 4321 ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta 4381 tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat 4441 aggggttccg cgcacatttc cccgaaaagt gccacctgac gtctaagaaa ccattattat 4501 catgacatta acctataaaa ataggcgtat cacgaggcc // LOCUS PTRIX2-RE9h-Hyg 8950 bp ds-DNA circular SYN 05-DEC-2016 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS . SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 8950) AUTHORS . TITLE Direct Submission JOURNAL Exported Jan 18, 2018 from SnapGene Viewer 4.0.6 http://www.snapgene.com FEATURES Location/Qualifiers source 1..8950 /organism="synthetic DNA construct" /mol_type="other DNA" rep_origin complement(6..461) /direction=LEFT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind 603..619 /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" promoter 626..644 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" CDS 1481..3124 /codon_start=1 /label=PpyRE9h Luciferase /translation="MEDAKNIKKGPAPFYPLEDGTAGEQLHKAMKRYALVPGTIAFTDA HIEVNITYAEYFEMSVRLAEAMKRYGLNTNHRIVVCSENSLQFFMPVLGALFIGVAVAP ANDIYNERELLNSMNISQPTVVFVSKKGLQKILNVQKKLPIIQKIIIMDSKTDYQGFQS MYTFVTSHLPPGFNEYDFVPESFDRDKTIALIMNSSGSTGLPKGVALPHRALCVRFSHA RDPIFGNQIAPDTAILSVVPFHHGFGMFTTLGYLICGFRVVLMYRFEEELFLRSLQDYK IQTALLVPTLFSFLAKSTLIDKYDLSNLHEIASGGAPLSKEVGEAVAKGFHLPGIRQGY GLTETTSAILVTPIGDDKPGAVGKVVPFFEAKVVDLDTGKTLGVNQRGELCVRGPMIMS GYVNNPEATNALIDKDGWLHSGDIAYWDEDEHFFIVDRLKSLIKYKGYQVAPAELESIL LQHPNIRDAGVAGLPDDDAGELPAAVVVLEHGKTMTEKEIVDYVASQVTTAKKLRGGVV FVDEVPKGLTGKLDARKIREILIKAKKGGKI" CDS 3869..4894 /codon_start=1 /gene="aph(4)-Ia" /product="aminoglycoside phosphotransferase from E. coli" /label=HygR /note="confers resistance to hygromycin" /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK E" promoter complement(6765..6783) /label=T3 promoter /note="promoter for bacteriophage T3 RNA polymerase" primer_bind complement(6804..6820) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" protein_bind 6828..6844 /label=lac operator /bound_moiety="lac repressor encoded by lacI" /note="The lac repressor binds to the lac operator to inhibit transcription in E. coli. This inhibition can be relieved by adding lactose or isopropyl-beta-D-thiogalactopyranoside (IPTG)." promoter complement(6852..6882) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 6897..6918 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." rep_origin complement(7206..7794) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" CDS complement(7965..8825) /codon_start=1 /gene="bla" /product="beta-lactamase" /label=AmpR /note="confers resistance to ampicillin, carbenicillin, and related antibiotics" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS LIKHW" promoter complement(8826..8930) /gene="bla" /label=AmpR promoter ORIGIN 1 cacctaaatt gtaagcgtta atattttgtt aaaattcgcg ttaaattttt gttaaatcag 61 ctcatttttt aaccaatagg ccgaaatcgg caaaatccct tataaatcaa aagaatagac 121 cgagataggg ttgagtgttg ttccagtttg gaacaagagt ccactattaa agaacgtgga 181 ctccaacgtc aaagggcgaa aaaccgtcta tcagggcgat ggcccactac gtgaaccatc 241 accctaatca agttttttgg ggtcgaggtg ccgtaaagca ctaaatcgga accctaaagg 301 gagcccccga tttagagctt gacggggaaa gccggcgaac gtggcgagaa aggaagggaa 361 gaaagcgaaa ggagcgggcg ctagggcgct ggcaagtgta gcggtcacgc tgcgcgtaac 421 caccacaccc gccgcgctta atgcgccgct acagggcgcg tcccattcgc cattcaggct 481 gcgcaactgt tgggaagggc gatcggtgcg ggcctcttcg ctattacgcc agctggcgaa 541 agggggatgt gctgcaaggc gattaagttg ggtaacgcca gggttttccc agtcacgacg 601 ttgtaaaacg acggccagtg aattgtaata cgactcacta tagggcgaat tggagctcga 661 cgtcggtgca ccctgggata tttgcgcacc cacctttccc gtcgcaatcg gcgccgtccg 721 aacgcggaaa tgttccgaga ataatacggc tgggctgcgc cgcgcgttcc ccgcgtgaac 781 caactattgt gcgatgtgca gcatcctatt gtggtgcttt accgcacacc agtttccttg 841 cggggcttat ttggtgtgtt ggatttttaa ttttttgatg attcggagga attttttgca 901 atttttgggt acccctaaaa aaattttgga gtttccgggg ttcgatccaa cttgtgcttt 961 tttaaccgtc catttgtgtc tagtacatca tggagcggta ttctctccgg tggcccaggt 1021 ggcgtcggaa ggaatcggtg tccctcagga tgtttggttt ttcgtttttt tttttgcggt 1081 cttttgtttc gcatcgccgt atcgtgtgcg gcgcgacctt ctttttggct cattggtcag 1141 tgaggtttat aataaagtgg atcatttggg tttttggccc tttgcggcgc acccgcggaa 1201 tgggggaggg attttcatat aattgtagtg tacacattgt gtgtgggtcg caaaatgtca 1261 tgccgtgtgc aatgtttatt tgcgtgctgt gcgtgtcctt ttgtgattcc accacttgtg 1321 tgtattttac cttatcgtct ctccctcgcg ttggctgtgc gtgaattgtc tgtcgcattc 1381 gtgggaaatc aaaaggggaa tcaacgtgtt cttttttttt cagaagggga gtcaaagcgc 1441 cgtgtggatg ccaagtctgt gcaagagttc tagtggatcc atggaggacg ccaagaacat 1501 caagaaggga ccagccccct tctaccccct ggaggacggc acagccggcg agcagctgca 1561 caaggccatg aagcggtacg ccctggtgcc aggcaccatc gccttcaccg acgcccacat 1621 cgaggtgaac atcacctacg ccgagtactt cgagatgagc gtgcggctgg ccgaggccat 1681 gaagcggtac ggcctgaaca ccaaccaccg gatcgtggtg tgcagcgaga acagcctgca 1741 gttcttcatg cccgtgctgg gagccctgtt catcggcgtg gccgtggccc cagccaacga 1801 catctacaac gagcgggagc tgctgaacag catgaacatc agccagccca ccgtggtgtt 1861 cgtgagcaag aagggcctgc agaagatcct gaatgtgcag aagaagctgc ccatcatcca 1921 gaagatcatc atcatggaca gcaagaccga ttaccagggc ttccagagca tgtacacctt 1981 cgtgaccagc cacctgcccc caggcttcaa cgagtacgac ttcgtgcccg agagcttcga 2041 ccgggacaag accatcgccc tgatcatgaa cagcagcggc agcaccggcc tgcccaaggg 2101 cgtggccctg ccccaccggg ccctgtgcgt gcggttcagc cacgccagag accccatctt 2161 cggcaaccag atcgcccccg acaccgccat cctgagcgtg gtgcccttcc accacggctt 2221 cggcatgttc accaccctgg gctacctgat ctgcggcttc cgggtggtgc tgatgtacag 2281 gttcgaggag gagctgttcc tgcggagcct gcaggactac aagatccaga ccgccctgct 2341 ggtgcccacc ctgttcagct tcctggccaa gagcaccctg atcgacaagt acgacctgag 2401 caacctgcac gagatcgcct ctggcggagc cccactgagc aaggaggtgg gcgaggccgt 2461 ggccaagggc ttccacctgc caggcatccg gcagggctac ggcctgaccg agaccaccag 2521 cgccatcctg gtgaccccca tcggcgacga caagcccgga gccgtgggca aggtggtgcc 2581 cttcttcgag gccaaggtgg tggacctgga caccggcaag accctgggcg tgaaccagag 2641 aggcgagctg tgcgtgagag gccccatgat catgagcggc tacgtgaaca accccgaggc 2701 caccaacgcc ctgatcgaca aggacggctg gctgcacagc ggcgacatcg cctactggga 2761 cgaggacgag cacttcttca tcgtggaccg gctgaagagc ctgatcaagt acaagggcta 2821 ccaggtggcc ccagccgagc tggagagcat cctgctgcag caccccaaca tccgggacgc 2881 cggagtggcc ggactgcccg acgacgacgc cggagagctg ccagccgccg tggtggtgct 2941 ggagcacggc aagaccatga ccgagaagga gatcgtggac tacgtggcca gccaggtgac 3001 caccgccaag aagctgagag gcggcgtggt gttcgtggac gaggtgccca agggcctgac 3061 cggcaagctg gacgccagaa agatccggga gatcctgatc aaggccaaga agggcggcaa 3121 gatctgactc gagccattta cgactccaag gcaacgctgc agaacaacct gccgaaggag 3181 cgccgcttct tcaagattgt gtcgtggtac gacaacgagt ggggatactc ccaccgcgtg 3241 gtggaccttg tacgccacat ggcctcgaag gatcgttcgg caaggttgta ggcgtggcga 3301 tgacttcagg tctttctttt gcgaataggg atcttataat acacgatgcg tgtcccgtga 3361 tgatcgttac cggtgctgcc acgatccaat tgacacagcg tcaagagcaa aacaatttta 3421 cttttccctt taaggacaac aacaaaaaaa tatataactt ttttttcttt tttttttttt 3481 tttgaaatta tatttatggt catctttggg aaacaaaaag cagcaattta atgatgcgga 3541 aggatgagtg aaataatgtt taatcaatgt acgaggattt ggggtattgc aaggaaaatg 3601 tagatgattt aattgggtgt gtgatgcagc ttgtggtaat ttttgctcac ttcccttttt 3661 gccacatctt tttagttttt ctgcttttct ttccccatta ttccacttgt ctctcttttc 3721 ccacgtttcc tgcacgaatg cagaaagtga tatttttact ttgaaagcca tctaccaaca 3781 acaattacat tgaacagaat ttgggattgc gaattaattc ctgcagcccc gcgtgtggcc 3841 tcgaacaccg agcgaccctg cagccaatat gaaaaagcct gaactcaccg cgacgtctgt 3901 cgagaagttt ctgatcgaaa agttcgacag cgtctccgac ctgatgcagc tctcggaggg 3961 cgaagaatct cgtgctttca gcttcgatgt aggagggcgt ggatatgtcc tgcgggtaaa 4021 tagctgcgcc gatggtttct acaaagatcg ttatgtttat cggcactttg catcggccgc 4081 gctcccgatt ccggaagtgc ttgacattgg ggaattcagc gagagcctga cctattgcat 4141 ctcccgccgt gcacagggtg tcacgttgca agacctgcct gaaaccgaac tgcccgctgt 4201 tctgcagccg gtcgcggagg ccatggatgc gatcgctgcg gccgatctta gccagacgag 4261 cgggttcggc ccattcggac cgcaaggaat cggtcaatac actacatggc gtgatttcat 4321 atgcgcgatt gctgatcccc atgtgtatca ctggcaaact gtgatggacg acaccgtcag 4381 tgcgtccgtc gcgcaggctc tcgatgagct gatgctttgg gccgaggact gccccgaagt 4441 ccggcacctc gtgcacgcgg atttcggctc caacaatgtc ctgacggaca atggccgcat 4501 aacagcggtc attgactgga gcgaggcgat gttcggggat tcccaatacg aggtcgccaa 4561 catcttcttc tggaggccgt ggttggcttg tatggagcag cagacgcgct acttcgagcg 4621 gaggcatccg gagcttgcag gatcgccgcg gctccgggcg tatatgctcc gcattggtct 4681 tgaccaactc tatcagagct tggttgacgg caatttcgat gatgcagctt gggcgcaggg 4741 tcgatgcgac gcaatcgtcc gatccggagc cgggactgtc gggcgtacac aaatcgcccg 4801 cagaagcgcg gccgtctgga ccgatggctg tgtagaagta ctcgccgata gtggaaaccg 4861 acgccccagc actcgtccga gggcaaagga atagggggat cgatccggaa caacctgccg 4921 aaggagcgcc gcttcttcaa gattgtgtcg tggtacgaca acgagtgggg atactcccac 4981 cgcgtggtgg accttgtacg ccacatggcc tcgaaggatc gttcggcaag gttgtaggcg 5041 tggcgatgac ttcaggtctt tcttttgcga atagggatct tataatacac gatgcgtgtc 5101 ccgtgatgat cgttaccggt gctgccacga tccaagtgac acagtgtcaa gagcaaaaca 5161 attttacttt tccctttaag gacaacaaca aaaaaatata taactttttt ttcttttttt 5221 ttttttgaaa ttatatttat ggtcatcttt gggaaacaaa aagcagcaat ttaatgatgc 5281 ggaaggatga gtgaaataat gtttaatcaa tgtacgagga tttggggtat tgcaaggaaa 5341 atgtagatga tttagttggg tgtatgatgc agcttgtggt aattttttac aactttgcgt 5401 tattggaaga ttttggttgt tcagtttctt tttgtttata actgactgtt ttaatttagt 5461 gtgatttttt ttttttaagg atttaagctg tttttctttt tttttctttt tcttcactga 5521 tggtgtttaa ggtgtagttt agtgtttgtg cattatgcgt cgtagtgttg ttgttctttg 5581 gactttttcg aacaatactg ggcattatag cccacctgtg tatatgccgc tagagtatgc 5641 tagccgtgtg ggtacctatt attactactg ctgctgctgc tgccactcct acatatgcgc 5701 acaagtacac tatatatatg cgaatacacc tgtgtgtatt atgctatgtg tgtgtctgtg 5761 tgggtgtgta ggtgatgggg tagtggttgt attatagtat agtatatata gtagtatacg 5821 gcaaaaggca aagacacaag cacggcgagt acgacgaaca aacccatttc acacatgagt 5881 gtgcgccaca cacgcgactg ctcctttttt ttctgccttg ccaccacccc atatgtatat 5941 tatatatata tatatataag ggtgtgtgtg tggtgagaga aaataggaaa tgattgcgtg 6001 tgtgtcgccg ccacgaacct gtcagatgct ttcttacggt attttgttgc atgcacccgc 6061 gtacacacaa tatatacaca cacaaaaaaa aacaccgata tgatggcgag tgtgcgattc 6121 tccttttatt gtctttttcc tctttttctg tgctctgctg tgtgccaata aaacacacga 6181 gagtagagaa acgggggaaa aagtagtaaa taaatggagg actgcttcgc agctgccctg 6241 ccgcactgtt tttgctgctg ggttcgcaac gcgatgctgt tgagagtgaa atcacatcac 6301 aaaaagcacc cctcagactg gtgtgaagtg tgaagtatgt gtacaaatgc gtgtgtgtgt 6361 gtatacacat tgacgaatag cataccaggg aaaaaaggga ggacactctc tttcgtgcgt 6421 gctgaactct gtgtgtgtgt gctgccgttt tgtttgcagc ttgaaatgct tggttgaata 6481 gaaatcggta cttgattgtt gccgcatttc gaggaatttc atccaaaaaa aaaaagaggt 6541 caacaacaac caactgacaa actttgtgtt ggacctgtgt atgtggtggt gtggtggggg 6601 aagagaaaca cacacacaga caaacacaaa caccatgtca ttttggcatg cgtgtgcgtg 6661 cgtgcgtgcg ttgaatatcc tccacacaca aatatatata tataataata ccacaacaac 6721 atttcaaagg atcatcaagg cgcgccggta cccagctttt gttcccttta gtgagggtta 6781 atttcgagct tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg ttatccgctc 6841 acaattccac acaacatacg agccggaagc ataaagtgta aagcctgggg tgcctaatga 6901 gtgagctaac tcacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg 6961 tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg 7021 cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg 7081 gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga 7141 aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg 7201 gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag 7261 aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc 7321 gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg 7381 ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt 7441 cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc 7501 ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc 7561 actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg 7621 tggcctaact acggctacac tagaaggaca gtatttggta tctgcgctct gctgaagcca 7681 gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc 7741 ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat 7801 cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt 7861 ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt 7921 tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc 7981 agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc 8041 gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata 8101 ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg 8161 gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc 8221 cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct 8281 acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa 8341 cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt 8401 cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca 8461 ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac 8521 tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca 8581 atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt 8641 tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc 8701 actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca 8761 aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata 8821 ctcatactct tcctttttca atattattga agcatttatc agggttattg tctcatgagc 8881 ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc 8941 cgaaaagtgc //