LOCUS Exported 5686 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION BM3R1-B3 gate. ACCESSION . VERSION . KEYWORDS pBM3R1-B3 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 5686) AUTHORS Nielsen AA, Der BS, Shin J, Vaidyanathan P, Paralanov V, Strychalski EA, Ross D, Densmore D, Voigt CA TITLE Genetic circuit design automation. JOURNAL Science. 2016 Apr 1;352(6281):aac7341. doi: 10.1126/science.aac7341. PUBMED 27034378 REFERENCE 2 (bases 1 to 5686) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..5686 /organism="synthetic DNA construct" /mol_type="other DNA" rep_origin complement(308..852) /direction=LEFT /label=p15A ori /note="Plasmids containing the medium-copy-number p15A origin of replication can be propagated in E. coli cells that contain a second plasmid with the ColE1 origin. " primer_bind complement(388..407) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(941..1756) /codon_start=1 /gene="aph(3')-Ia" /product="aminoglycoside phosphotransferase" /label=KanR /note="confers resistance to kanamycin in bacteria or G418 (Geneticin(R)) in eukaryotes" /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF" primer_bind 1664..1683 /label=Kan-R /note="Kanamycin resistance gene, reverse primer" primer_bind 1748..1767 /label=pENTR-R /note="pENTR vectors, reverse primer" promoter 1878..1906 /label=tac promoter /note="strong E. coli promoter; hybrid between the trp and lac UV5 promoters" protein_bind 1912..1931 /label=lac operator (symmetric) /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). The symmetric lac operator was optimized for tight binding of lac repressor." misc_RNA 2737..2786 /label=sTRSV HHRz /note="hammerhead ribozyme from the tobacco ringspot virus satellite RNA (Khvorova et al., 2003)" CDS 2836..3555 /codon_start=1 /product="enhanced YFP" /label=EYFP /note="mammalian codon-optimized" /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KFICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKLHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL EFVTAAGITLGMDELYK" primer_bind complement(2881..2902) /label=EGFP-N /note="EGFP, reverse primer" primer_bind complement(3142..3161) /label=EXFP-R /note="For distinguishing EGFP variants, reverse primer" primer_bind 3489..3510 /label=EGFP-C /note="EGFP, forward primer" CDS 3703..4785 /codon_start=1 /gene="lacI" /product="lac repressor" /label=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" primer_bind complement(3722..3741) /label=LacI-R /note="LacI, reverse primer" CDS 4804..5427 /codon_start=1 /gene="tetR from transposon Tn10" /product="tetracycline repressor TetR" /label=TetR /note="TetR binds to the tetracycline operator tetO to inhibit transcription. This inhibition can be relieved by adding tetracycline or doxycycline." /translation="MSRLDKSKVINSALELLNEVGIEGLTTRKLAQKLGVEQPTLYWHV KNKRALLDALAIEMLDRHHTHFCPLEGESWQDFLRNNAKSFRCALLSHRDGAKVHLGTR PTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEDQEHQVAKEERETPTT DSMPPLLRQAIELFDHQGAEPAFLFGLELIICGLEKQLKCESGS" primer_bind complement(4877..4896) /label=Tet-R /note="Tetracycline resistance gene, reverse primer" primer_bind complement(5610..5629) /label=pBRrevBam /note="pBR322 vectors, tet region, downstream of BamHI, reverse primer" ORIGIN 1 gcttcctcgc tcactgactc gctgcacgag gcagacctca gcgctagcgg agtgtatact 61 ggcttactat gttggcactg atgagggtgt cagtgaagtg cttcatgtgg caggagaaaa 121 aaggctgcac cggtgcgtca gcagaatatg tgatacagga tatattccgc ttcctcgctc 181 actgactcgc tacgctcggt cgttcgactg cggcgagcgg aaatggctta cgaacggggc 241 ggagatttcc tggaagatgc caggaagata cttaacaggg aagtgagagg gccgcggcaa 301 agccgttttt ccataggctc cgcccccctg acaagcatca cgaaatctga cgctcaaatc 361 agtggtggcg aaacccgaca ggactataaa gataccaggc gtttcccctg gcggctccct 421 cgtgcgctct cctgttcctg cctttcggtt taccggtgtc attccgctgt tatggccgcg 481 tttgtctcat tccacgcctg acactcagtt ccgggtaggc agttcgctcc aagctggact 541 gtatgcacga accccccgtt cagtccgacc gctgcgcctt atccggtaac tatcgtcttg 601 agtccaaccc ggaaagacat gcaaaagcac cactggcagc agccactggt aattgattta 661 gaggagttag tcttgaagtc atgcgccggt taaggctaaa ctgaaaggac aagttttggt 721 gactgcgctc ctccaagcca gttacctcgg ttcaaagagt tggtagctca gagaaccttc 781 gaaaaaccgc cctgcaaggc ggttttttcg ttttcagagc aagagattac gcgcagacca 841 aaacgatctc aagaagatca tcttattaag gggtctgacg ctcagtggaa cgaaaaatca 901 atctaaagta tatatgagta aacttggtct gacagttacc ttagaaaaac tcatcgagca 961 tcaaatgaaa ctgcaattta ttcatatcag gattatcaat accatatttt tgaaaaagcc 1021 gtttctgtaa tgaaggagaa aactcaccga ggcagttcca taggatggca agatcctggt 1081 atcggtctgc gattccgact cgtccaacat caatacaacc tattaatttc ccctcgtcaa 1141 aaataaggtt atcaagtgag aaatcaccat gagtgacgac tgaatccggt gagaatggca 1201 aaagcttatg catttctttc cagacttgtt caacaggcca gccattacgc tcgtcatcaa 1261 aatcactcgc atcaaccaaa ccgttattca ttcgtgattg cgcctgagcg agacgaaata 1321 cgcgatcgct gttaaaagga caattacaaa caggaatcga atgcaaccgg cgcaggaaca 1381 ctgccagcgc atcaacaata ttttcacctg aatcaggata ttcttctaat acctggaatg 1441 ctgttttccc ggggatcgca gtggtgagta accatgcatc atcaggagta cggataaaat 1501 gcttgatggt cggaagaggc ataaattccg tcagccagtt tagtctgacc atctcatctg 1561 taacatcatt ggcaacgcta cctttgccat gtttcagaaa caactctggc gcatcgggct 1621 tcccatacaa tcgatagatt gtcgcacctg attgcccgac attatcgcga gcccatttat 1681 acccatataa atcagcatcc atgttggaat ttaatcgcgg cctcgagcaa gacgtttccc 1741 gttgaatatg gctcataaca ccccttgtat tactgtttat gtaagcagac agttttattg 1801 ttcatgatga tatattttta tcttgtgcaa tgtacatcag agattttgag acacaaggag 1861 aacgatcgtt ggctgtgttg acaattaatc atcggctcgt ataatgtgtg gaattgtgag 1921 cgctcacaat tagactgtcg ccggatgtgt atccgacctg acgatggccc aaaagggccg 1981 aaacagtcct ctacaaataa ttttgtttaa ccaaacgagg ccgggaggat ggaaagcacc 2041 ccgaccaaac agaaagcaat ttttagcgca agcctgctgc tgtttgcaga acgtggtttt 2101 gatgcaacca ccatgccgat gattgcagaa aatgcaaaag ttggtgcagg caccatttat 2161 cgctatttca aaaacaaaga aagcctggtg aacgaactgt ttcagcagca tgttaatgaa 2221 tttctgcagt gtattgaaag cggtctggca aatgaacgtg atggttatcg tgatggcttt 2281 catcacattt ttgaaggtat ggtgaccttt accaaaaatc atccgcgtgc actgggtttt 2341 atcaaaaccc atagccaggg cacctttctg accgaagaaa gccgtctggc atatcagaaa 2401 ctggttgaat ttgtgtgcac cttttttcgt gaaggtcaga aacagggtgt gattcgtaat 2461 ctgccggaaa atgcactgat tgcaattctg tttggcagct ttatggaagt gtatgaaatg 2521 atcgagaacg attatctgag cctgaccgat gaactgctga ccggtgttga agaaagcctg 2581 tgggcagcac tgagccgtca gagctaactc ggtaccaaat tccagaaaag agacgctttc 2641 gagcgtcttt tttcgttttg gtcctactaa tccgcgtgat aggtctgatt cgttaccaat 2701 tgacggaatg aacgttcatt ccgataatgc tagcagctgt caccggatgt gctttccggt 2761 ctgatgagtc cgtgaggacg aaacagcctc tacaaataat tttgtttaat actagagaaa 2821 gaggggaaat actagatggt gagcaagggc gaggagctgt tcaccggggt ggtgcccatc 2881 ctggtcgagc tggacggcga cgtaaacggc cacaagttca gcgtgtccgg cgagggcgag 2941 ggcgatgcca cctacggcaa gctgaccctg aagttcatct gcaccacagg caagctgccc 3001 gtgccctggc ccaccctcgt gaccaccttc ggctacggcc tgcaatgctt cgcccgctac 3061 cccgaccaca tgaagctgca cgacttcttc aagtccgcca tgcccgaagg ctacgtccag 3121 gagcgcacca tcttcttcaa ggacgacggc aactacaaga cccgcgccga ggtgaagttc 3181 gagggcgaca ccctggtgaa ccgcatcgag ctgaagggca tcgacttcaa ggaggacggc 3241 aacatcctgg ggcacaagct ggagtacaac tacaacagcc acaacgtcta tatcatggcc 3301 gacaagcaga agaacggcat caaggtgaac ttcaagatcc gccacaacat cgaggacggc 3361 agcgtgcagc tcgccgacca ctaccagcag aacaccccaa tcggcgacgg ccccgtgctg 3421 ctgcccgaca accactacct tagctaccag tccgccctga gcaaagaccc caacgagaag 3481 cgcgatcaca tggtcctgct ggagttcgtg accgccgccg ggatcactct cggcatggac 3541 gagctgtaca agtaactcgg taccaaattc cagaaaagag gcctcccgaa aggggggcct 3601 tttttcgttt tggtccaatg gcggcgcgcc atcgaatggc gcaaaacctt tcgcggtatg 3661 gcatgatagc gcccggaaga gagtcaattc agggtggtga atatgaaacc agtaacgtta 3721 tacgatgtcg cagagtatgc cggtgtctct tatcagaccg tttcccgcgt ggtgaaccag 3781 gccagccacg tttctgcgaa aacgcgggaa aaagtggaag cggcgatggc ggagctgaat 3841 tacattccca accgcgtggc acaacaactg gcgggcaaac agtcgttgct gattggcgtt 3901 gccacctcca gtctggccct gcacgcgccg tcgcaaattg tcgcggcgat taaatctcgc 3961 gccgatcaac tgggtgccag cgtggtggtg tcgatggtag aacgaagcgg cgtcgaagcc 4021 tgtaaagcgg cggtgcacaa tcttctcgcg caacgcgtca gtgggctgat cattaactat 4081 ccgctggatg accaggatgc cattgctgtg gaagctgcct gcactaatgt tccggcgtta 4141 tttcttgatg tctctgacca gacacccatc aacagtatta ttttctccca tgaggacggt 4201 acgcgactgg gcgtggagca tctggtcgca ttgggtcacc agcaaatcgc gctgttagcg 4261 ggcccattaa gttctgtctc ggcgcgtctg cgtctggctg gctggcataa atatctcact 4321 cgcaatcaaa ttcagccgat agcggaacgg gaaggcgact ggagtgccat gtccggtttt 4381 caacaaacca tgcaaatgct gaatgagggc atcgttccca ctgcgatgct ggttgccaac 4441 gatcagatgg cgctgggcgc aatgcgcgcc attaccgagt ccgggctgcg cgttggtgcg 4501 gatatctcgg tagtgggata cgacgatacc gaagatagct catgttatat cccgccgtta 4561 accaccatca aacaggattt tcgcctgctg gggcaaacca gcgtggaccg cttgctgcaa 4621 ctctctcagg gccaggcggt gaagggcaat cagctgttgc cagtctcact ggtgaaaaga 4681 aaaaccaccc tggcgcccaa tacgcaaacc gcctctcccc gcgcgttggc cgattcatta 4741 atgcagctgg cacgacaggt ttcccgactg gaaagcgggc agtgataatc caggaggaaa 4801 aaaatgtcca gattagataa aagtaaagtg attaacagcg cattagagct gcttaatgag 4861 gtcggaatcg aaggtttaac aacccgtaaa ctcgcccaga agctaggtgt agagcagcct 4921 acattgtatt ggcatgtaaa aaataagcgg gctttgctcg acgccttagc cattgagatg 4981 ttagataggc accatactca cttttgccct ttagaagggg aaagctggca agatttttta 5041 cgtaataacg ctaaaagttt tagatgtgct ttactaagtc atcgcgatgg agcaaaagta 5101 catttaggta cacggcctac agaaaaacag tatgaaactc tcgaaaatca attagccttt 5161 ttatgccaac aaggtttttc actagagaat gcattatatg cactcagcgc tgtggggcat 5221 tttactttag gttgcgtatt ggaagatcaa gagcatcaag tcgctaaaga agaaagggaa 5281 acacctacta ctgatagtat gccgccatta ttacgacaag ctatcgaatt atttgatcac 5341 caaggtgcag agccagcctt cttattcggc cttgaattga tcatatgcgg attagaaaaa 5401 caacttaaat gtgaaagtgg gtcctaataa ttggtaacga atcagacaat tgacggctcg 5461 agggagtagc atagggtttg cagaatccct gcttcgtcca tttgacaggc acattatgca 5521 tcgatgataa gctgtcaaac atgagcagat cctctacgcc ggacgcatcg tggccggcat 5581 caccggcgcc acaggtgcgg ttgctggcgc ctatatcgcc gacatcaccg atggggaaga 5641 tcgggctcgc cacttcgggc tcatgagcaa atattttatc tgaggt //