LOCUS Exported 9065 bp ds-DNA circular SYN 18-DEC-2015 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS BiFC1 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 9065) AUTHORS . TITLE Direct Submission JOURNAL Exported Friday, Aug 4, 2017 from SnapGene 3.2.0 http://www.snapgene.com FEATURES Location/Qualifiers source 1..9065 /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" CDS 321..2503 /codon_start=1 /note="CmLEU2" /translation="AGIRPWIQSSLVGVKKEV*DQVLWHHH**LGLVKLQD**NKNLHL TRNILRGYLLN*RTVCYPFHLLSLTVVITQHINTQVV*MFPLHTLKGNLC*WH*KILH* VLVLHVPLHLWNHRMFYMLWVPMMPWLILPLDSVLVDSPLKQKLTMLFKQLMKELIS*E RCLHYGKWFKKVLT*TLSNGVDIKVFCFFVICVVMCNYRV*SNFGNCTFVHVSEPDE*Y VRIVEE*LTPVTACVGSAAGFNFFFLVHYKNI**TKKIRGTQAMKFFTQEFFFLHP*FL YSLPKPTMSVKTKTITILPGDHVGTEIVNEAIKVLEAIEAATPYQKIHFDFKHHLIGGA AIDATGVPLPDDALESAKKSDAVLLGAVGGPKWGTGAVRPEQGLLKIRKELNLYANIRP CNFASDSLLELSPLRPEVVKGTNLIIVRELVGGIYFGDREEQEESEDKQTAWDTEKYTV DEVTRITRMAAFMALQHNPPLPIWSLDKANVLASSRLWRRTVDKVISEEFPTLSVQHQL IDSAAMILIQNPTKLNGIIITSNMFGDIISDEASVIPGSLGLLPSASLASLPDTNTAFG LYEPCHGSAPDLPANKVNPIATILSAASMLRLSLDCVKEAEALEEAVKQVLDKGIRTAD LRGTSSATEVGDAIVEAVTKILKEKA*VKYTQNLTNIINTK*TPPKYFFLKMYRLF*KL R*FNIGNFSHRFTWIFGNKSI*SNGSLT*F*LA" promoter complement(2596..2614) /note="T7 promoter" /note="promoter for bacteriophage T7 RNA polymerase" primer_bind complement(2621..2637) /note="M13 fwd" /note="common sequencing primer, one of multiple similar variants" misc_feature 2741..3858 /note="IntegSeq" /note="Integration Sequence (a 1104-bp fragment of genomic DNA on chromosome 1, between genes XOG1 and HOL1. 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 3868..4170 /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 4519..5313 /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 5568..7007 /note="MET3prom" CDS 7014..7529 /codon_start=1 /note="yEmVenus 1-172" /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK LICTTGKLPVPWPTLVTTLGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIKA NFKIRHNIE" misc_feature 7536..7565 /note="Linker" misc_feature 7568..7598 /note="MCS" misc_feature 7590..7592 /note="STOP CODON" terminator 7621..8134 /note="CaACT1term" rep_origin 8296..8884 /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 ctagtaacgg ccgccagtgt gctggaattc gcccttggat ccaatcatca ctggtggggg 361 tcaagaaaga ggtttgagat caggtacttt ggcaccacca ttagtagctg ggtttggtga 421 agctgcaaga ttaatgaaac aagaatctgc atttgacaag aaacatattg agaggttatc 481 tactaaattg aagaacggtt tgttatccat tccatctact cagtttaacg gttgtaataa 541 cccaacatat caatacccag gttgtgtgaa tgtttccttt gcatacattg aaggggaatc 601 tttgttaatg gcattgaaag atattgcatt gagttctggt tctgcatgta cctctgcatc 661 tttggaacca tcgtatgttt tacatgcttt gggtgccgat gatgccttgg ctcattcttc 721 cattagattc ggtattggta gattcaccac tgaagcagaa gttgactatg ttattcaagc 781 aattaatgaa agagttgatt tcttgagaaa gatgtctcca ttatgggaaa tggttcaaga 841 aggtattgac ttgaactcta tcgaatggag tggacattaa ggttttttgt ttttttgtaa 901 tttgtgttgt tatgtgtaat tatcgtgtat agagtaattt cggaaattgt actttcgttc 961 atgtatctga accagatgaa taatacgttc gtattgttga agaatgacta actcctgtaa 1021 cggcgtgcgt gggttccgca gccgggttca attttttttt tcttgttcac tataaaaata 1081 tataataaac gaaaaaaatc cgagggactc aggctatgaa atttttcact caagaatttt 1141 ttttccttca tccttaattc ctttattcac tacccaaacc cacaatgtca gttaaaacta 1201 aaactattac aattttacca ggtgatcacg tcggtaccga aattgtcaat gaagctatca 1261 aagttttaga agcaattgaa gctgccactc cttaccaaaa aatccatttc gatttcaaac 1321 atcatttaat cggtggtgct gccattgatg ccactggtgt cccattgcca gatgacgcac 1381 ttgaaagtgc caaaaaatct gatgccgtct tattgggtgc tgttggtgga ccaaaatggg 1441 ggactggcgc cgttcgtcca gaacaaggtt tattaaagat tcgtaaagaa ttgaaccttt 1501 atgctaacat cagaccatgt aactttgcca gtgactcctt attggaatta tccccattga 1561 gaccagaagt cgttaaaggt acaaacttga tcattgttcg tgaattagtt ggtggtattt 1621 atttcggtga tcgtgaagaa caagaagaaa gtgaagataa acaaaccgca tgggatactg 1681 aaaaatacac cgtcgatgaa gtcaccagaa tcacccgtat ggctgccttc atggctttac 1741 aacataaccc accattacca atctggtcat tggataaggc taatgtttta gcttcttcta 1801 gattatggag aagaactgtc gataaagtga tttctgaaga gttcccaact ttgtctgttc 1861 aacatcaatt gattgactcc gccgccatga ttttaattca aaacccaacc aaattgaatg 1921 gtataatcat cacttctaac atgtttggtg atatcatttc cgatgaagct tcagttattc 1981 caggttcctt gggtttatta ccatctgcat ctttggcttc attgccagat accaacaccg 2041 cttttggtct ttatgaacca tgtcacggtt ctgctcctga tttaccagcc aataaagtta 2101 acccaattgc tactatttta tctgctgctt ctatgttgag attatctttg gattgtgtga 2161 aagaagctga agctttagaa gaagccgtga aacaagtctt ggataaaggt atcagaactg 2221 cagatttaag aggtaccagt tcagctactg aagttggtga cgcgattgtc gaagctgtta 2281 ctaaaatctt aaaagaaaaa gcttaagtaa agtatacaca aaatttaacg aatattatta 2341 atacaaaatg aacaccacct aaatactttt ttttaaaaat gtaccgactc ttctaaaaat 2401 tgcgataatt caatatcgga aatttctctc accggtttac ttggatcttc ggaaataaat 2461 ctatttgaag taatggtagc ctcacttgat tctgattggc tctttctaga catgggtagg 2521 taagggcgaa ttctgcagat atccatcaca ctggcggccg gccgctcgag catgcatcta 2581 gagggcccaa ttcgccctat agtgagtcgt attacaattc actggccgtc gttttacaac 2641 gtcgtgactg ggaaaaccct ggcgttaccc aacttaatcg ccttgcagca catccccctt 2701 tcgccagctg gcgtaatagc gaagaggccc gcaccgatcg accagatcaa aatgctagaa 2761 caaattacgt ctgcatcaac agtcatcttc cttatttgca taaatgctaa aatggcaaac 2821 ctgtaaaccg tttgaaaagc aaattgagat agttagctcg gttttttttt cacgtggcca 2881 taatccaaca aaaaaaagga atttaagggg ggagagaagg aggcttgaat ttgtaccaca 2941 gagcgagcaa gaaattaata atatacttgg cttagctgaa atgagaatga aaaagcaaat 3001 ttcaagtgaa acaccactac tagccaagta aattggccaa tttgctctcg tgctttaatt 3061 tcctaacaag cttttgagac cgaaaaaaat caaccaatga tgatggaaaa atacttccgg 3121 gagtcaattt atccataatt tcacgtcgtt ttttttttgc acatggcaaa tatattggaa 3181 tcctgagatt ttcttggagt cgtgtacaat aacaccaggc gggaacatgc cgttatgatc 3241 cagcatatgg cggccgccat atgtgttcaa atattgaaac cgtgaaacca gtttgataca 3301 gctgatattg tgtgctaaaa aaagatgagc accgaaagag tctttagttt attgccaagt 3361 aaagggtcaa agaataccta ttctttatga acacagactt gctctaatgg tgaattagtg 3421 tggagtaata gagaaggagg cgaaacaaaa ataattggtt aaacactcga atgtcacaca 3481 atcctaaata ggttaaattc agttaccgcc gtgatgcggg aacaaagccg agcggtaata 3541 tttttggatt ctatgttttt ttggtgtata ttgcacgact tggtgccaac aactggtgtt 3601 ccaacgcaca aaactaggtt caactggata ctgttttcta gaaattattt gcggcaattc 3661 atgattacgg tcctaaattt gaaataaaag tttccaatta tgtgtcttga caatttccat 3721 tgggttcaaa aagatctaga atgtatgtaa ttacgcaaca atttgcatgc aaagaaataa 3781 aaacaaacct aaaatccaac tgctgttgtt ctggtgatga tgtagtaaaa taagagcacg 3841 gccaagaaca aggtgattta cgtacggcag tttaaggttt acacctataa aagagagagc 3901 cgttatcgtc tgtttgtgga tgtacagagt gatattattg acacgccggg gcgacggatg 3961 gtgatccccc tggccagtgc acgtctgctg tcagataaag tctcccgtga actttacccg 4021 gtggtgcata tcggggatga aagctggcgc atgatgacca ccgatatggc cagtgtgccg 4081 gtctccgtta tcggggaaga agtggctgat ctcagccacc gcgaaaatga catcaaaaac 4141 gccattaacc tgatgttctg gggaatataa atgtcaggca tgagattatc aaaaaggatc 4201 ttcacctaga tccttttcac gtagaaagcc agtccgcaga aacggtgctg accccggatg 4261 aatgtcagct actgggctat ctggacaagg gaaaacgcaa gcgcaaagag aaagcaggta 4321 gcttgcagtg ggcttacatg gcgatagcta gactgggcgg ttttatggac agcaagcgaa 4381 ccggaattgc cagctggggc gccctctggt aaggttggga agccctgcaa agtaaactgg 4441 atggctttct cgccgccaag gatctgatgg cgcaggggat caagctctga tcaagagaca 4501 ggatgaggat cgtttcgcat gattgaacaa gatggattgc acgcaggttc tccggccgct 4561 tgggtggaga ggctattcgg ctatgactgg gcacaacaga caatcggctg ctctgatgcc 4621 gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc 4681 ggtgccctga atgaactgca agacgaggca gcgcggctat cgtggctggc cacgacgggc 4741 gttccttgcg cagctgtgct cgacgttgtc actgaagcgg gaagggactg gctgctattg 4801 ggcgaagtgc cggggcagga tctcctgtca tctcaccttg ctcctgccga gaaagtatcc 4861 atcatggctg atgcaatgcg gcggctgcat acgcttgatc cggctacctg cccattcgac 4921 caccaagcga aacatcgcat cgagcgagca cgtactcgga tggaagccgg tcttgtcgat 4981 caggatgatc tggacgaaga gcatcagggg ctcgcgccag ccgaactgtt cgccaggctc 5041 aaggcgagca tgcccgacgg cgaggatctc gtcgtgaccc atggcgatgc ctgcttgccg 5101 aatatcatgg tggaaaatgg ccgcttttct ggattcatcg actgtggccg gctgggtgtg 5161 gcggaccgct atcaggacat agcgttggct acccgtgata ttgctgaaga gcttggcggc 5221 gaatgggctg accgcttcct cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc 5281 gccttctatc gccttcttga cgagttcttc tgaattatta acgcttacaa tttcctgatg 5341 cggtattttc tccttacgca tctgtgcggt atttcacacc gcatacaggt ggcacttttc 5401 ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc 5461 cgctcatgag acaataaccc tgataaatgc ttcaataata gcacgtgagg agggccacca 5521 tggccaagtt gaccagtgcc gttccggtgc tcaccgcgcg cgacgtcttc caattagagg 5581 ttgtggatgc aaaattgaaa aaaaaatgac tattggtagc gtgttctatt tttcttatgc 5641 gctgaaaaac tacgaacaat tgtctattcc aagcctgtgg ctttttttgg tgattcacaa 5701 tacataaatt ggcaaatacc tcttctatag actttaacct ttgatgaaat ggtatggata 5761 aaccactttt gtggtcaggg tatatatatt tttattccaa tctggccttt ataaattcaa 5821 atttccaagg ggactctgct aaaccttctg atataactat aactctaata atggctgtaa 5881 aaagtgtgaa aaaaagatga tattcacaaa aaagataaag gggatttcat attgggtatc 5941 tagatttaaa catcgtattt caacaatgaa actagaatca atacgaaatc cagggcacaa 6001 ctgtgttcat tcccttggga attatactga gtaatgggaa gaatgtgcaa aaaatacccg 6061 tgtcttatat aatttccatt tgtgtagggc ttaaaagtgg acaacttcag gttacgaaac 6121 cacgaaaatg tctaaacgtt tgatcaaatt cgtaactcat ggaaaatcac agcaacaaat 6181 taaggatgga tatatccggt acttacaaat aaaaacacta aagaagtata aattttgaac 6241 tgaaattcta tagaaaatct ttttttttgg gtaaaattat gcgattgtgg ctcatagtaa 6301 cgtaatcata cagttcaaca aacatcattt tgtcaaagga tttctaatcc tattcttttt 6361 ctacttggct tttgagtata tatatatata tatatacatc ctttgttcat catcatgttc 6421 gattgaattt ttttcttggt tgttattaga ttgtctactt gagtttttct tggtgcaaat 6481 tgtggtgttt ttctaacaaa tttttcaaca tttaaagtat ttctggaact tttctacaag 6541 gaacgacata gatccattaa tgcgcccctc taaaacaata cccataatgt cttcttaagt 6601 atgcatctgc ttataaatgt ttctggattt actttagtta gaatgtcgac acgacaatga 6661 actattgctg tagatcacgt gcgatcactg aattatgagt ttctttttat ttttcttttt 6721 gggctttttt ttttgcaatt ttttttttcc acctttgcca attgcgtagt cacatataat 6781 gtatacaatt tttgcaactc tgacactata attgatccaa taataaagta tataatagat 6841 catcaagtat acgtaatctc cccaaaaaaa aaattgcata aaataaaatc tataaacctt 6901 ttttagattt ttagttgttc cttttttctt tctattcttc aacttccaag tgttgtcact 6961 ttcttcctgt taactatatt taaaagtaaa taccctcccc agaaaactcc ggaatgtcta 7021 aaggtgaaga attattcact ggtgttgtcc caattttggt tgaattagat ggtgatgtta 7081 atggtcacaa attttctgtc tccggtgaag gtgaaggtga tgctacttac ggtaaattga 7141 ccttaaaatt gatttgtact actggtaaat tgccagttcc atggccaacc ttagtcacta 7201 ctttaggtta tggtttgcaa tgttttgcta gatacccaga tcatatgaaa caacatgact 7261 ttttcaagtc tgccatgcca gaaggttatg ttcaagaaag aactattttt ttcaaagatg 7321 acggtaacta caagaccaga gctgaagtca agtttgaagg tgatacctta gttaatagaa 7381 tcgaattaaa aggtattgat tttaaagaag atggtaacat tttaggtcac aaattggaat 7441 acaactataa ctctcacaat gtttacatca ctgctgacaa acaaaagaat ggtatcaaag 7501 ctaacttcaa aattagacac aacattgaag ctagcggagg tggaggttct ggtggaggtg 7561 gttcaagcct agggacccgg gggcgcgcct aaacgcgtag tacttaaccg cgccgtcgac 7621 gagtgaaatt ctggaaatct ggaaatctgg ttttgtattc ttgttattct tctttttgtt 7681 attacatata taacttgtta cttttttaaa aaaatctttg tttattttat aaatatataa 7741 aactaaattt aagaaaaaga gaaaaatgtt ttatttgaga gattgaaatt ttacttgaat 7801 ttagcttagc ttttataaag tattattatg taaaaaaaca aaacaaatat acattaaaaa 7861 gttaagacta taaaatagcc acccaaggca tttctatatc ttgttgttgt tgttttcatc 7921 ttctgtatca gaggaactta ttttattatt ttcgtcacgg gtattttctc ttgtttgatg 7981 attcatccca ttcattccat cataaaattc tagaaggacc acctttgatt gtaaatagta 8041 ataattacca cccttattta attatttatt taacttattt atttatttat tatacatata 8101 tacaaatcta ataaagtgaa aatctccccc ttcacacgtg gccgaggagc aggactgaca 8161 cgtgctaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct 8221 catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa 8281 gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa 8341 aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc 8401 gaaggtaact ggcttcagca gagcgcagat accaaatact gtccttctag tgtagccgta 8461 gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct 8521 gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg 8581 atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag 8641 cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc 8701 cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg 8761 agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt 8821 tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg 8881 gaaaaacgcc agcaacgcgg cctttttacg gttcctgggc ttttgctggc cttttgctca 8941 catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg cctttgagtg 9001 agctgatacc gctcgccgca gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc 9061 ggaag //