LOCUS Exported 9743 bp ds-DNA circular SYN 18-DEC-2015 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS BiFC3 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 9743) AUTHORS . TITLE Direct Submission JOURNAL Exported Friday, Aug 4, 2017 from SnapGene 3.2.0 http://www.snapgene.com FEATURES Location/Qualifiers source 1..9743 /organism="synthetic DNA construct" /mol_type="other DNA" protein_bind 107..128 /bound_moiety="E. coli catabolite activator protein" /note="CAP binding site" /note="CAP binding activates transcription in the presence of cAMP." promoter 143..173 /note="lac promoter" /note="promoter for the E. coli lac operon" protein_bind 181..197 /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)." primer_bind 205..221 /note="M13 rev" /note="common sequencing primer, one of multiple similar variants" promoter 239..257 /note="SP6 promoter" /note="promoter for bacteriophage SP6 RNA polymerase" gene 306..3508 /note="CdARG4" misc_feature 3606..4463 /note="IntegSeq" /note="Integration Sequence (a 850-bp sequence located between RXT3 and ORF19.3569 on chromosome 2. After restriction with NotI, the linearized vector will be targeted to this genomic site for integration.) See Stynen et al. Nucleic Acids Research, 2010, Vol. 38, No. 19 e184 " CDS 4612..4914 /codon_start=1 /gene="ccdB" /product="CcdB, a bacterial toxin that poisons DNA gyrase" /note="ccdB" /note="Plasmids containing the ccdB gene cannot be propagated in standard E. coli strains." /translation="QFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDKV SRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI" CDS 5263..6057 /codon_start=1 /gene="aph(3')-II (or nptII)" /product="aminoglycoside phosphotransferase from Tn5" /note="NeoR/KanR" /note="confers resistance to neomycin, kanamycin, and G418 (Geneticin(R))" /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF" promoter 6312..7751 /note="MET3prom" CDS 7758..7958 /codon_start=1 /note="yEmVenus 173-238" /translation="MDGGVQLADHYQQNTPIGDGPVLLPDNHYLSYQSKLSKDPNEKRD HMVLLEFVTAAGITHGMDELYK" misc_feature 7965..7994 /note="Linker" misc_feature 7997..8027 /note="MCS" misc_feature 8019..8021 /note="STOP CODON" terminator 8062..8575 /note="CaACT1term" rep_origin 8974..9562 /direction=RIGHT /note="ori" /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" ORIGIN 1 agcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc 61 acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc 121 tcactcatta ggcaccccag gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa 181 ttgtgagcgg ataacaattt cacacaggaa acagctatga ccatgattac gccaagctat 241 ttaggtgaca ctatagaata ctcaagctat gcatcaagct tggtaccgag ctcggatcca 301 ctagtttctg taccgcaaat gtatcgactt tttcgatggt gtgttattgt tgttttgttg 361 cgaaatataa tagcttctgc tttatatcct ttgtgttgat ccttcccctc ctttcatact 421 tgtaaaatta caattggtat tttcaaccaa tattaccaca cgtcatgcac taactcaaat 481 gggcaaaaaa aagagagata ggaattacgt cattttatta tgtttttacg tggactagaa 541 ggttaagata atttctggtt attttgttga taattgagtt ttgttcgacc tccaccaatc 601 tttaaactct cgttgagtat tcattttaat gattttttga aatcagaagg ggtgtttcta 661 acccaggatt gttttattga gaggttgaaa catcaatgtg ttagtttatt gatttgggtt 721 ggtttaatag aaaaatatca gtaggtgttc aataattgta gtttattttc ttggctaaaa 781 ttgttgcgaa aaattgacaa aataataaca aaaagtattt tagatgctaa aacataatag 841 taggaatatc tacctgaacc atttacattc aacaactggc aagaaccttc ctaataattt 901 gcaagagtaa tcacaattaa accaaaatca aagtttaccg tgtatatagt tggacagaaa 961 gctgtcacat tggtttaata catagttgaa tatgtatgca tgaaacaaca acaatacaat 1021 tacagctcat gatccttcaa ttaatttcaa cattcaaaca acataacacc ttgtgtaaat 1081 gaggttatga gtacatgtga acaacaaatc ttatgactaa actcgggtcg tgtacttttg 1141 acgattgagt catttttacc aattaggcaa ttaatttttg ctaaattttc attaaccgcg 1201 ctggccatcc caacaaacaa tataaaagct gacaacgaaa aaaaaaaaaa aaaaaaaaaa 1261 actccaattc tgaaaaattt ttcttcttct tcatactaaa caaaacaact ataaagataa 1321 tcaattaata atgtcacaac aacctagtga aaataaatta tggggtggtc gattcactgg 1381 cgccactgat ccattaatgg atttatacaa tgcttcatta ccttatgata aagttatgta 1441 tgatgcagat ttaactggta ctaaagtata tactcaagga ttgaacaaat taggattaat 1501 taccactgaa gaattgaatc tgattcatca aggtttagaa caaattagac aagaatggca 1561 tgataataaa tttattatta aagctggtga tgaagatatt catactgcta atgaacgtag 1621 attaggagaa atcattggga aaaacatttc tggtaaagtt catactggta gatccagaaa 1681 tgatcaagtt gctactgata tgagaatttt tgttcgtgaa tcattactta atttaagtaa 1741 gattttacat caattcatta cagcaattct tgaacgagct cataaagaaa ttgatgtatt 1801 aatgcctggt tatactcatt tacaaaaagc tcaacctatt agatgggctc attggttaag 1861 ttcatatgca acttatttca ctgaagatta taaacgttta caagaaatca ttactcgtgt 1921 taatcaatct ccattaggat ctggtgcttt agctggtcat ccctacggta ttgatcgtga 1981 atttttagct aaaggattag gatttgatgg tgttattggg aattctttaa ctgcagtttc 2041 tgatcgtgat tttgttgttg aaagtttatt ttggagtaca ttatttatga atcatatttc 2101 tcgattttca gaagatttaa ttatttattc tagtggtgaa tttggattta taaagttagc 2161 tgatgcttac tctactggtt catcattaat gccacaaaag aaaaatcctg attcattaga 2221 attattacgt gggaaaagtg gcagagtgtt tggtcagtta tctgggtttt taatgtcaat 2281 taaatccatt ccatcaacct ataataaaga tatgcaagaa gataaagaac cattatttga 2341 tgcattgact accgtagaac attcaatttt aattgctact ggtgtgattt ctactttact 2401 gattgataaa caaaatatgg aaaaagcttt aactatggat atgttagcta ctgatttagc 2461 agattattta gttagaaaag gtgttccatt tagagaaact catcatattt ctggtgaatg 2521 tgttagaaaa gctgaagaag aaaaattgtc gggaattgac caattatcat ttgaacaatt 2581 tcaacaaatt gattcacgtt ttgaaaaaga tgttatggaa acttttgatt ttgaagctag 2641 tgtggaaaga agagatgcta ttggtggtac tgctaagagc gctgttttaa aacagttaga 2701 aaatttgaaa tcaattttaa attaatttat aaataatcat atagtattca atgttttgtt 2761 ttgttttttt tttttttttt atggaaatgt acaaacagta tataatcaat taatagaaag 2821 ataccctgta ttccattgta tcgcctttgg gatgtctata ataccaactt tcaagaatga 2881 gaaattaaac aacaatttct ttcaccaatc aactcgtcta tattcaataa actcattctg 2941 gtcttttcaa atatttaatg ttatcctgat caaacttaca aatcaaaaat gtatcaaaat 3001 cattcattag ttcttcttct tttgttatct ttgtcactat atttctctcc atccagacca 3061 aaaatacttc cattgtgaag atcgtctgaa tcataatcag caactacctc ttccatttca 3121 gagaattcag atttttcaaa ataggtatca cgggccaagt acacacaaga caaacgatac 3181 gagtgctggg atatttcttt caatgtatca ctaaattcgt tgagatttaa agagagccct 3241 ttgactgctt tcattacttt cctcacatca tgagcatcca cattcaaatt tcctgtagcc 3301 ctttattctt agttttcttt tcttattctt ttcttcgtac attgaaatca caaatggtga 3361 tatcaaacga aggtcacact gacttatgtc taatacctcc aatgatgata ctggtaactt 3421 aaccaggtct actcttttca aacgatttct cattagtgtc aatttacgta ggttcttcaa 3481 cctttcaaac gatgcaaata gcttttgtaa cggccgccag tgtgctggaa ttgcccttaa 3541 gggcgaattc tgcagatatc catcacactg gcggccggcc gctcgagcat gcatctagag 3601 ggcccttgaa gaattggaag aagttggcag ttaaaaaaca atgggaaaga tttaatcttt 3661 tttttcttct ttcaaaaatt tttaaccgct gatgatatat taatctctct ccaccactct 3721 cgttaaacac cacaattatt aataataaga tatctctcct actaaatatt tgttagtaag 3781 tatattgttg attgattggt tgggttaaaa attttttttt tcataaagaa aataatgaaa 3841 gtgggaaaaa tttaaacccc acattacata ctcaacacaa cactacacac acgcatttaa 3901 caatcatcat cagttggaat atattttaca aacaattacg attgtttaat gaaaactatt 3961 tctgttatta ttatttaacc tttttttttt cttgttgggt tgaccctttt tctattaact 4021 atctaactac cacataacat ttgaagttga actattgaac cttaatttgc ggccgcaaat 4081 ttaatcttct ttcttcttcc accttttttt tatttttttg aaatgggggg tggtgggaac 4141 tttaggcgtg ttaaatttgt cacatctgtt tcacttgctt ctattcgtat aattttaatt 4201 gtcttgtgag atcaatcttg ttcttgctct tgttttttct tgctttgatt aactaccatt 4261 attattagta tgcaatgctg tttattacca tttcataact tatattataa ctcattgatt 4321 tcaatacctg ctcaacaatg agtcgttatt attactgtgc tctgtattct gtactgctgc 4381 acgtacgtta tgtccattga agacaagcaa agaaagacca acattcaatt gtatctgctt 4441 accagagttg tttacctctg ttaccaatat ttggaaatta attttttggt ggtggacgtt 4501 tcttcctaaa acacctagat gtgctagtat aattgcacaa ccagcacgca cactgcgtct 4561 ggccggtcct ggactacatt ttgtctctaa cattcgtagc tgcacgtacg gcagtttaag 4621 gtttacacct ataaaagaga gagccgttat cgtctgtttg tggatgtaca gagtgatatt 4681 attgacacgc cggggcgacg gatggtgatc cccctggcca gtgcacgtct gctgtcagat 4741 aaagtctccc gtgaacttta cccggtggtg catatcgggg atgaaagctg gcgcatgatg 4801 accaccgata tggccagtgt gccggtctcc gttatcgggg aagaagtggc tgatctcagc 4861 caccgcgaaa atgacatcaa aaacgccatt aacctgatgt tctggggaat ataaatgtca 4921 ggcatgagat tatcaaaaag gatcttcacc tagatccttt tcacgtagaa agccagtccg 4981 cagaaacggt gctgaccccg gatgaatgtc agctactggg ctatctggac aagggaaaac 5041 gcaagcgcaa agagaaagca ggtagcttgc agtgggctta catggcgata gctagactgg 5101 gcggttttat ggacagcaag cgaaccggaa ttgccagctg gggcgccctc tggtaaggtt 5161 gggaagccct gcaaagtaaa ctggatggct ttctcgccgc caaggatctg atggcgcagg 5221 ggatcaagct ctgatcaaga gacaggatga ggatcgtttc gcatgattga acaagatgga 5281 ttgcacgcag gttctccggc cgcttgggtg gagaggctat tcggctatga ctgggcacaa 5341 cagacaatcg gctgctctga tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt 5401 ctttttgtca agaccgacct gtccggtgcc ctgaatgaac tgcaagacga ggcagcgcgg 5461 ctatcgtggc tggccacgac gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa 5521 gcgggaaggg actggctgct attgggcgaa gtgccggggc aggatctcct gtcatctcac 5581 cttgctcctg ccgagaaagt atccatcatg gctgatgcaa tgcggcggct gcatacgctt 5641 gatccggcta cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact 5701 cggatggaag ccggtcttgt cgatcaggat gatctggacg aagagcatca ggggctcgcg 5761 ccagccgaac tgttcgccag gctcaaggcg agcatgcccg acggcgagga tctcgtcgtg 5821 acccatggcg atgcctgctt gccgaatatc atggtggaaa atggccgctt ttctggattc 5881 atcgactgtg gccggctggg tgtggcggac cgctatcagg acatagcgtt ggctacccgt 5941 gatattgctg aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc 6001 gccgctcccg attcgcagcg catcgccttc tatcgccttc ttgacgagtt cttctgaatt 6061 attaacgctt acaatttcct gatgcggtat tttctcctta cgcatctgtg cggtatttca 6121 caccgcatac aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt 6181 tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat 6241 aatagcacgt gaggagggcc accatggcca agttgaccag tgccgttccg gtgctcaccg 6301 cgcgcgacgt cttccaatta gaggttgtgg atgcaaaatt gaaaaaaaaa tgactattgg 6361 tagcgtgttc tatttttctt atgcgctgaa aaactacgaa caattgtcta ttccaagcct 6421 gtggcttttt ttggtgattc acaatacata aattggcaaa tacctcttct atagacttta 6481 acctttgatg aaatggtatg gataaaccac ttttgtggtc agggtatata tatttttatt 6541 ccaatctggc ctttataaat tcaaatttcc aaggggactc tgctaaacct tctgatataa 6601 ctataactct aataatggct gtaaaaagtg tgaaaaaaag atgatattca caaaaaagat 6661 aaaggggatt tcatattggg tatctagatt taaacatcgt atttcaacaa tgaaactaga 6721 atcaatacga aatccagggc acaactgtgt tcattccctt gggaattata ctgagtaatg 6781 ggaagaatgt gcaaaaaata cccgtgtctt atataatttc catttgtgta gggcttaaaa 6841 gtggacaact tcaggttacg aaaccacgaa aatgtctaaa cgtttgatca aattcgtaac 6901 tcatggaaaa tcacagcaac aaattaagga tggatatatc cggtacttac aaataaaaac 6961 actaaagaag tataaatttt gaactgaaat tctatagaaa atcttttttt ttgggtaaaa 7021 ttatgcgatt gtggctcata gtaacgtaat catacagttc aacaaacatc attttgtcaa 7081 aggatttcta atcctattct ttttctactt ggcttttgag tatatatata tatatatata 7141 catcctttgt tcatcatcat gttcgattga atttttttct tggttgttat tagattgtct 7201 acttgagttt ttcttggtgc aaattgtggt gtttttctaa caaatttttc aacatttaaa 7261 gtatttctgg aacttttcta caaggaacga catagatcca ttaatgcgcc cctctaaaac 7321 aatacccata atgtcttctt aagtatgcat ctgcttataa atgtttctgg atttacttta 7381 gttagaatgt cgacacgaca atgaactatt gctgtagatc acgtgcgatc actgaattat 7441 gagtttcttt ttatttttct ttttgggctt ttttttttgc aatttttttt ttccaccttt 7501 gccaattgcg tagtcacata taatgtatac aatttttgca actctgacac tataattgat 7561 ccaataataa agtatataat agatcatcaa gtatacgtaa tctccccaaa aaaaaaattg 7621 cataaaataa aatctataaa ccttttttag atttttagtt gttccttttt tctttctatt 7681 cttcaacttc caagtgttgt cactttcttc ctgttaacta tatttaaaag taaataccct 7741 ccccagaaaa ctccggaatg gatggtggtg ttcaattagc tgaccattat caacaaaata 7801 ctccaattgg tgatggtcca gtcttgttac cagacaacca ttacttatcc tatcaatcta 7861 aattatccaa agatccaaac gaaaagagag accacatggt cttgttagaa tttgttactg 7921 ctgctggtat tacccatggt atggatgaat tgtacaaagc tagcggaggt ggaggttctg 7981 gtggaggtgg ttcaagccta gggacccggg ggcgcgccta aacgcgtagt acttaagatc 8041 ctgaagacgg gatcgttcga agagtgaaat tctggaaatc tggaaatctg gttttgtatt 8101 cttgttattc ttctttttgt tattacatat ataacttgtt acttttttaa aaaaatcttt 8161 gtttatttta taaatatata aaactaaatt taagaaaaag agaaaaatgt tttatttgag 8221 agattgaaat tttacttgaa tttagcttag cttttataaa gtattattat gtaaaaaaac 8281 aaaacaaata tacattaaaa agttaagact ataaaatagc cacccaaggc atttctatat 8341 cttgttgttg ttgttttcat cttctgtatc agaggaactt attttattat tttcgtcacg 8401 ggtattttct cttgtttgat gattcatccc attcattcca tcataaaatt ctagaaggac 8461 cacctttgat tgtaaatagt aataattacc acccttattt aattatttat ttaacttatt 8521 tatttattta ttatacatat atacaaatct aataaagtga aaatctcccc cttcacccgg 8581 tgtggtccgg gacgacgtga ccctgttcat cagcgcggtc caggaccagg tggtgccgga 8641 caacaccctg gcctgggtgt gggtgcgcgg cctggacgag ctgtacgccg agtggtcgga 8701 ggtcgtgtcc acgaacttcc gggacgcctc cgggccggcc atgaccgaga tcggcgagca 8761 gccgtggggg cgggagttcg ccctgcgcga cccggccggc aactgcgtgc acttcgtggc 8821 cgaggagcag gactgacacg tgctaaaact tcatttttaa tttaaaagga tctaggtgaa 8881 gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc 8941 gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat 9001 ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga 9061 gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt 9121 ccttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata 9181 cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac 9241 cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg 9301 ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg 9361 tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag 9421 cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct 9481 ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc 9541 aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt tcctgggctt 9601 ttgctggcct tttgctcaca tgttctttcc tgcgttatcc cctgattctg tggataaccg 9661 tattaccgcc tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga 9721 gtcagtgagc gaggaagcgg aag //