LOCUS Exported 8679 bp ds-DNA circular SYN 06-JUN-2018 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS pET-SOMA SOURCE synthetic DNA construct ORGANISM recombinant plasmid REFERENCE 1 (bases 1 to 8679) AUTHORS Krysan Lab TITLE Direct Submission JOURNAL Exported Jun 6, 2018 from SnapGene 2.8.3 http://www.snapgene.com FEATURES Location/Qualifiers source 1..8679 /organism="recombinant plasmid" /mol_type="other DNA" CDS 599..1459 /codon_start=1 /gene="bla" /product="beta-lactamase" /note="AmpR" /note="confers resistance to ampicillin, carbenicillin, and related antibiotics" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS LIKHW" rep_origin 1630..2218 /direction=RIGHT /note="ori" /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" CDS complement(3647..4729) /codon_start=1 /gene="lacI" /product="lac repressor" /note="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)." /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR ALADSLMQLARQVSRLESGQ" promoter complement(4730..4807) /gene="lacI" /note="lacI promoter" promoter 5120..5138 /note="T7 promoter" /note="promoter for bacteriophage T7 RNA polymerase" protein_bind 5139..5163 /bound_moiety="lac repressor encoded by lacI" /note="lac operator" /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)." RBS 5178..5200 /note="efficient ribosome binding site from bacteriophage T7 gene 10 (Olins and Rangwala, 1989)" CDS 5208..5534 /codon_start=1 /gene="trxA" /product="E. coli thioredoxin" /note="TrxA" /translation="MSDKIIHLTDDSFDTDVLKADGAILVDFWAEWCGPCKMIAPILDE IADEYQGKLTVAKLNIDQNPGTAPKYGIRGIPTLLLFKNGEVAATKVGALSKGQLKEFL DANLA" CDS 5556..5573 /codon_start=1 /product="6xHis affinity tag" /note="6xHis" /translation="HHHHHH" CDS 5607..5651 /codon_start=1 /product="affinity and epitope tag derived from pancreatic ribonuclease A" /note="S-Tag" /translation="KETAAAKFERQHMDS" CDS 5664..8511 /codon_start=1 /product="FRET Biosensor" /note="FRET Sensor CDS" /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KLLCTTGKLPVPWPTLVTTLGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIK ANFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALFKDPNEKRDHMVLL EFLTAAGITEGMNELYLEKFSQEQIGENIVCRVICTTGQIPIRDLSADISQVLKEKRSI KKVWTFGRNPACDYHLGNISRLSNKHFQILLGEDGNLLLNDISTNGTWLNGQKVEKNSN QLLSQGDEITVGVGVESDILSLVIFINDKFKQCLEQNKVDRGTSAGGSAGGSAGGSAGG SAGGSGSAGGSAGGSTSAGGSAGGSAGGSAGGSAGGSGSAGGSAGGSTSAGGSAGGSAG GSAGGSAGGSGSAGGSAGGSTSAGGSAGGSAGGSAGGSAGGSGSAGGSAGGSTSAGGSA GGSAGGSAGGSAGGSGSAGGSAGGSTSAGGSAGGSAGGSAGGSAGGSGSAGGSAGGSTS AGGSAGGSAGGSAGGSAGGSGSAGGSAGGSTSAGGSAGGSAGGSAGGSAGGSGPGSKKM FWRSASWSASRTASQVPEGDEQSLNIPCAISSGPSRRCPAAPLTPRDHHNSKARACLPP LQPLAISRRSLDEWPGGRMVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYG KLTLKFICTTGKLPVPWPTLVTTLSWGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIF FKDDGNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYISGNVYITADKQK NGIKANFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDH MVLLEFLTAAGITLGMDEL" CDS 8524..8541 /codon_start=1 /product="6xHis affinity tag" /note="6xHis" /translation="HHHHHH" terminator 8608..8655 /note="T7 terminator" /note="transcription terminator for bacteriophage T7 RNA polymerase" ORIGIN 1 tggcgaatgg gacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg 61 cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc 121 ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg 181 gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc 241 acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt 301 ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc 361 ttttgattta taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta 421 acaaaaattt aacgcgaatt ttaacaaaat attaacgttt acaatttcag gtggcacttt 481 tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta 541 tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtat 601 gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt gccttcctgt 661 ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg 721 agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt ttcgccccga 781 agaacgtttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg tattatcccg 841 tattgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga atgacttggt 901 tgagtactca ccagtcacag aaaagcatct tacggatggc atgacagtaa gagaattatg 961 cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga caacgatcgg 1021 aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa ctcgccttga 1081 tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca ccacgatgcc 1141 tgcagcaatg gcaacaacgt tgcgcaaact attaactggc gaactactta ctctagcttc 1201 ccggcaacaa ttaatagact ggatggaggc ggataaagtt gcaggaccac ttctgcgctc 1261 ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc gtgggtctcg 1321 cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag ttatctacac 1381 gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga taggtgcctc 1441 actgattaag cattggtaac tgtcagacca agtttactca tatatacttt agattgattt 1501 aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac 1561 caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa 1621 aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc 1681 accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt 1741 aactggcttc agcagagcgc agataccaaa tactgtcctt ctagtgtagc cgtagttagg 1801 ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc 1861 agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt 1921 accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga 1981 gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct 2041 tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg 2101 cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca 2161 cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa 2221 cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt 2281 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga 2341 taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga 2401 gcgcctgatg cggtattttc tccttacgca tctgtgcggt atttcacacc gcatatatgg 2461 tgcactctca gtacaatctg ctctgatgcc gcatagttaa gccagtatac actccgctat 2521 cgctacgtga ctgggtcatg gctgcgcccc gacacccgcc aacacccgct gacgcgccct 2581 gacgggcttg tctgctcccg gcatccgctt acagacaagc tgtgaccgtc tccgggagct 2641 gcatgtgtca gaggttttca ccgtcatcac cgaaacgcgc gaggcagctg cggtaaagct 2701 catcagcgtg gtcgtgaagc gattcacaga tgtctgcctg ttcatccgcg tccagctcgt 2761 tgagtttctc cagaagcgtt aatgtctggc ttctgataaa gcgggccatg ttaagggcgg 2821 ttttttcctg tttggtcact gatgcctccg tgtaaggggg atttctgttc atgggggtaa 2881 tgataccgat gaaacgagag aggatgctca cgatacgggt tactgatgat gaacatgccc 2941 ggttactgga acgttgtgag ggtaaacaac tggcggtatg gatgcggcgg gaccagagaa 3001 aaatcactca gggtcaatgc cagcgcttcg ttaatacaga tgtaggtgtt ccacagggta 3061 gccagcagca tcctgcgatg cagatcggaa cataatggtg cagggcgctg acttccgcgt 3121 ttccagactt tacgaaacac ggaaaccgaa gaccattcat gttgttgctc aggtcgcaga 3181 cgttttgcag cagcagtcgc ttcacgttcg ctcgcgtatc ggtgattcat tctgctaacc 3241 agtaaggcaa ccccgccagc ctagccgggt cctcaacgac aggagcacga tcatgcgcac 3301 ccgtggggcc gccatgccgg cgataatggc ctgcttctcg ccgaaacgtt tggtggcggg 3361 accagtgacg aaggcttgag cgagggcgtg caagattccg aataccgcaa gcgacaggcc 3421 gatcatcgtc gcgctccagc gaaagcggtc ctcgccgaaa atgacccaga gcgctgccgg 3481 cacctgtcct acgagttgca tgataaagaa gacagtcata agtgcggcga cgatagtcat 3541 gccccgcgcc caccggaagg agctgactgg gttgaaggct ctcaagggca tcggtcgaga 3601 tcccggtgcc taatgagtga gctaacttac attaattgcg ttgcgctcac tgcccgcttt 3661 ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg 3721 cggtttgcgt attgggcgcc agggtggttt ttcttttcac cagtgagacg ggcaacagct 3781 gattgccctt caccgcctgg ccctgagaga gttgcagcaa gcggtccacg ctggtttgcc 3841 ccagcaggcg aaaatcctgt ttgatggtgg ttaacggcgg gatataacat gagctgtctt 3901 cggtatcgtc gtatcccact accgagatgt ccgcaccaac gcgcagcccg gactcggtaa 3961 tggcgcgcat tgcgcccagc gccatctgat cgttggcaac cagcatcgca gtgggaacga 4021 tgccctcatt cagcatttgc atggtttgtt gaaaaccgga catggcactc cagtcgcctt 4081 cccgttccgc tatcggctga atttgattgc gagtgagata tttatgccag ccagccagac 4141 gcagacgcgc cgagacagaa cttaatgggc ccgctaacag cgcgatttgc tggtgaccca 4201 atgcgaccag atgctccacg cccagtcgcg taccgtcttc atgggagaaa ataatactgt 4261 tgatgggtgt ctggtcagag acatcaagaa ataacgccgg aacattagtg caggcagctt 4321 ccacagcaat ggcatcctgg tcatccagcg gatagttaat gatcagccca ctgacgcgtt 4381 gcgcgagaag attgtgcacc gccgctttac aggcttcgac gccgcttcgt tctaccatcg 4441 acaccaccac gctggcaccc agttgatcgg cgcgagattt aatcgccgcg acaatttgcg 4501 acggcgcgtg cagggccaga ctggaggtgg caacgccaat cagcaacgac tgtttgcccg 4561 ccagttgttg tgccacgcgg ttgggaatgt aattcagctc cgccatcgcc gcttccactt 4621 tttcccgcgt tttcgcagaa acgtggctgg cctggttcac cacgcgggaa acggtctgat 4681 aagagacacc ggcatactct gcgacatcgt ataacgttac tggtttcaca ttcaccaccc 4741 tgaattgact ctcttccggg cgctatcatg ccataccgcg aaaggttttg cgccattcga 4801 tggtgtccgg gatctcgacg ctctccctta tgcgactcct gcattaggaa gcagcccagt 4861 agtaggttga ggccgttgag caccgccgcc gcaaggaatg gtgcatgcaa ggagatggcg 4921 cccaacagtc ccccggccac ggggcctgcc accataccca cgccgaaaca agcgctcatg 4981 agcccgaagt ggcgagcccg atcttcccca tcggtgatgt cggcgatata ggcgccagca 5041 accgcacctg tggcgccggt gatgccggcc acgatgcgtc cggcgtagag gatcgagatc 5101 gatctcgatc ccgcgaaatt aatacgactc actatagggg aattgtgagc ggataacaat 5161 tcccctctag taataatttt gtttaacttt aagaaggaga tatacatatg agcgataaaa 5221 ttattcacct gactgacgac agttttgaca cggatgtact caaagcggac ggggcgatcc 5281 tcgtcgattt ctgggcagag tggtgcggtc cgtgcaaaat gatcgccccg attctggatg 5341 aaatcgctga cgaatatcag ggcaaactga ccgttgcaaa actgaacatc gatcaaaacc 5401 ctggcactgc gccgaaatat ggcatccgtg gtatcccgac tctgctgctg ttcaaaaacg 5461 gtgaagtggc ggcaaccaaa gtgggtgcac tgtctaaagg tcagttgaaa gagttcctcg 5521 acgctaacct ggccggttct ggttctggcc atatgcacca tcatcatcat cattcttctg 5581 gtctggtgcc acgcggttct ggtatgaaag aaaccgctgc tgctaaattc gaacgccagc 5641 acatggacag cccagaattc ggcatggtga gcaagggcga ggagctgttc accggggtgg 5701 tgcccatcct ggtcgagctg gacggcgacg taaacggcca caagttcagc gtgtccggcg 5761 agggcgaggg cgatgccacc tacggcaagc tgaccctgaa gcttctatgc accaccggca 5821 agctgcccgt gccctggccc accctcgtga ccaccctggg ctacggcctg cagtgcttcg 5881 cccgctaccc cgaccacatg aagcagcacg acttcttcaa gtccgccatg cccgaaggct 5941 acgtccagga gcgcaccatc ttcttcaagg acgacggcaa ctacaagacc cgcgccgagg 6001 tgaagttcga gggcgacacc ctggtgaacc gcatcgagct gaagggcatc gacttcaagg 6061 aggacggcaa catcctgggg cacaagctgg agtacaacta caacagccac aacgtctata 6121 tcaccgccga caagcagaag aacggcatca aggccaactt caagatccgc cacaacatcg 6181 aggacggcgg cgtgcagctc gccgaccact accagcagaa cacccccatc ggcgacggcc 6241 ccgtgctgct gcccgacaac cactacctga gctaccagtc cgccctgttc aaagacccca 6301 acgagaagcg cgatcacatg gtcctgctgg agttcctgac cgccgccggg atcactgagg 6361 gcatgaacga gctgtacctc gagaagtttt ctcaagaaca gatcggcgaa aacattgtgt 6421 gcagggtcat ttgtaccacg ggtcaaattc ccatccgaga tttgtcagct gatatttcac 6481 aagtgcttaa ggaaaaacga tccataaaga aagtttggac atttggtaga aacccagcct 6541 gtgactatca tttaggaaac atttcaagac tgtcaaataa gcatttccaa atactactag 6601 gagaagacgg taacctttta ttgaatgaca tttccactaa tgggacctgg ttaaatgggc 6661 aaaaagtcga gaagaacagc aatcagttac tgtcccaagg tgatgaaata accgttggtg 6721 taggcgtgga atcagatatt ttatctctgg tcattttcat aaacgacaaa tttaagcagt 6781 gcctggagca gaacaaagtt gatcgcggta ccagtgctgg tggtagtgct ggtggtagtg 6841 ctggtggtag tgctggtggt agtgctggtg gttccggcag tgctggtggt agtgctggtg 6901 gtagtaccag tgctggtggt agtgctggtg gtagtgctgg tggtagtgct ggtggtagtg 6961 ctggtggttc cggcagtgct ggtggtagtg ctggtggtag taccagtgct ggtggtagtg 7021 ctggtggtag tgctggtggt agtgctggtg gtagtgctgg tggttccggc agtgctggtg 7081 gtagtgctgg tggtagtacc agtgctggtg gtagtgctgg tggtagtgct ggtggtagtg 7141 ctggtggtag tgctggtggt tccggcagtg ctggtggtag tgctggtggt agtaccagtg 7201 ctggtggtag tgctggtggt agtgctggtg gtagtgctgg tggtagtgct ggtggttccg 7261 gcagtgctgg tggtagtgct ggtggtagta ccagtgctgg tggtagtgct ggtggtagtg 7321 ctggtggtag tgctggtggt agtgctggtg gttccggcag tgctggtggt agtgctggtg 7381 gtagtaccag tgctggtggt agtgctggtg gtagtgctgg tggtagtgct ggtggtagtg 7441 ctggtggttc cggcagtgct ggtggtagtg ctggtggtag taccagtgct ggtggtagtg 7501 ctggtggtag tgctggtggt agtgctggtg gtagtgctgg tggttccgga cctggttcta 7561 agaaaatgtt ttggcggtct gcctcttggt ctgcttcacg gactgcatca caagttcctg 7621 agggtgatga gcaaagcctg aacattccgt gtgctattag ttctgggccg agtcgaagat 7681 gtccagctgc tcctttgaca cctcgtgatc atcataacag caaggctaga gcttgtttgc 7741 caccattgca gcctcttgcc atttctagga ggagcttaga cgagtggcct ggcggccgca 7801 tggtgagcaa gggcgaggag ctgttcaccg gggtggtgcc catcctggtc gagctggacg 7861 gcgacgtaaa cggccacaag ttcagcgtgt ccggcgaggg cgagggcgat gccacctacg 7921 gcaagctgac cctgaagttc atctgcacca ccggcaagct gcccgtgccc tggcccaccc 7981 tcgtgaccac cctgtcctgg ggcgtgcagt gcttcgcccg ctaccccgac cacatgaagc 8041 agcacgactt cttcaagtcc gccatgcccg aaggctacgt ccaggagcgc accatcttct 8101 tcaaggacga cggcaactac aagacccgcg ccgaggtgaa gttcgagggc gacaccctgg 8161 tgaaccgcat cgagctgaag ggcatcgact tcaaggagga cggcaacatc ctggggcaca 8221 agctggagta caactacatc agcgggaacg tctatatcac cgccgacaag cagaagaacg 8281 gcatcaaggc caacttcaag atccgccaca acatcgagga cggcggcgtg cagctcgccg 8341 accactacca gcagaacacc cccatcggcg acggccccgt gctgctgccc gacaaccact 8401 acctgagcac ccagtccgcc ttaagcaaag accccaacga gaagcgcgat cacatggtcc 8461 tgctggagtt cttgaccgcc gccgggatca ctctcggcat ggacgagctg tctagacgtc 8521 gagcaccacc accaccacca ctgagatccg gctgctaaca aagcccgaaa ggaagctgag 8581 ttggctgctg ccaccgctga gcaataacta gcataacccc ttggggcctc taaacgggtc 8641 ttgaggggtt ttttgctgaa aggaggaact atatcggat //