LOCUS Exported 5829 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION PsrA-R1 gate. ACCESSION . VERSION . KEYWORDS pPsrA-R1 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 5829) 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 5829) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..5829 /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 2880..2929 /label=sTRSV HHRz /note="hammerhead ribozyme from the tobacco ringspot virus satellite RNA (Khvorova et al., 2003)" CDS 2979..3698 /codon_start=1 /product="enhanced YFP" /label=EYFP /note="mammalian codon-optimized" /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KFICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKLHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL EFVTAAGITLGMDELYK" primer_bind complement(3024..3045) /label=EGFP-N /note="EGFP, reverse primer" primer_bind complement(3285..3304) /label=EXFP-R /note="For distinguishing EGFP variants, reverse primer" primer_bind 3632..3653 /label=EGFP-C /note="EGFP, forward primer" CDS 3846..4928 /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(3865..3884) /label=LacI-R /note="LacI, reverse primer" CDS 4947..5570 /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(5020..5039) /label=Tet-R /note="Tetracycline resistance gene, reverse primer" primer_bind complement(5753..5772) /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 tagcgctgtc tgtacttgta tcagtacact gacgagtccc taaaggacga 1981 aacaccgcct ctacaaataa ttttgtttaa tttaattcgc ggaagcgcag agataagggg 2041 tatcatggca cagagcgaaa ccgttgaacg tattctggat gcagcagaac agctgtttgc 2101 agaacgtggt tttgcagaaa ccagcctgcg tctgattacc agcaaagccg gtgttaatct 2161 ggcagcagtg aattatcatt ttggcagcaa aaaagcactg attcaggcag tttttagccg 2221 ttttctgggt ccgttttgtg caagcctgga acgtgaactg gaacgtcgtc aggcacgtcc 2281 ggaacagaaa ccgagcctgg aagaactgct ggaaatgctg gttgaacagg cactggcagt 2341 tcagcctcgt agcaataatg atctgagcat ttttatgtgt ctgctgggtc tggcatttag 2401 ccagagccag ggtcatctgc gtcgttatct ggaagatatg tatggtaaag tgttccgtcg 2461 ttatatgctg ctggttaatg aagcagcacc gcgtgttccg cctctggaac tgttttggcg 2521 tgttcatttt atgctgggtg cagcagcatt tagcatgagc ggtattaaag cactgcgtgc 2581 aattgcagaa accgattttg gtattaacac cagcattgaa caggttatgc gtctgatggt 2641 tccgtttctg gcagcaggta tgcgtgcaga tagcggtgtt accgatgaag caatggcagc 2701 agcacagctg cgtccgcgta gcaaaaccag caccagcgca accaccgcaa aagcataatc 2761 cggcaattaa aaaagcggct aaccacgccg ctttttttac gtctgcatac ttgatcgaac 2821 gcttcaagga acaaacgttt gattgacagc tagctcagtc ctaggtagag tgctagcagc 2881 tgtcaccgga tgtgctttcc ggtctgatga gtccgtgagg acgaaacagc ctctacaaat 2941 aattttgttt aatactagag aaagagggga aatactagat ggtgagcaag ggcgaggagc 3001 tgttcaccgg ggtggtgccc atcctggtcg agctggacgg cgacgtaaac ggccacaagt 3061 tcagcgtgtc cggcgagggc gagggcgatg ccacctacgg caagctgacc ctgaagttca 3121 tctgcaccac aggcaagctg cccgtgccct ggcccaccct cgtgaccacc ttcggctacg 3181 gcctgcaatg cttcgcccgc taccccgacc acatgaagct gcacgacttc ttcaagtccg 3241 ccatgcccga aggctacgtc caggagcgca ccatcttctt caaggacgac ggcaactaca 3301 agacccgcgc cgaggtgaag ttcgagggcg acaccctggt gaaccgcatc gagctgaagg 3361 gcatcgactt caaggaggac ggcaacatcc tggggcacaa gctggagtac aactacaaca 3421 gccacaacgt ctatatcatg gccgacaagc agaagaacgg catcaaggtg aacttcaaga 3481 tccgccacaa catcgaggac ggcagcgtgc agctcgccga ccactaccag cagaacaccc 3541 caatcggcga cggccccgtg ctgctgcccg acaaccacta ccttagctac cagtccgccc 3601 tgagcaaaga ccccaacgag aagcgcgatc acatggtcct gctggagttc gtgaccgccg 3661 ccgggatcac tctcggcatg gacgagctgt acaagtaact cggtaccaaa ttccagaaaa 3721 gaggcctccc gaaagggggg ccttttttcg ttttggtcca atggcggcgc gccatcgaat 3781 ggcgcaaaac ctttcgcggt atggcatgat agcgcccgga agagagtcaa ttcagggtgg 3841 tgaatatgaa accagtaacg ttatacgatg tcgcagagta tgccggtgtc tcttatcaga 3901 ccgtttcccg cgtggtgaac caggccagcc acgtttctgc gaaaacgcgg gaaaaagtgg 3961 aagcggcgat ggcggagctg aattacattc ccaaccgcgt ggcacaacaa ctggcgggca 4021 aacagtcgtt gctgattggc gttgccacct ccagtctggc cctgcacgcg ccgtcgcaaa 4081 ttgtcgcggc gattaaatct cgcgccgatc aactgggtgc cagcgtggtg gtgtcgatgg 4141 tagaacgaag cggcgtcgaa gcctgtaaag cggcggtgca caatcttctc gcgcaacgcg 4201 tcagtgggct gatcattaac tatccgctgg atgaccagga tgccattgct gtggaagctg 4261 cctgcactaa tgttccggcg ttatttcttg atgtctctga ccagacaccc atcaacagta 4321 ttattttctc ccatgaggac ggtacgcgac tgggcgtgga gcatctggtc gcattgggtc 4381 accagcaaat cgcgctgtta gcgggcccat taagttctgt ctcggcgcgt ctgcgtctgg 4441 ctggctggca taaatatctc actcgcaatc aaattcagcc gatagcggaa cgggaaggcg 4501 actggagtgc catgtccggt tttcaacaaa ccatgcaaat gctgaatgag ggcatcgttc 4561 ccactgcgat gctggttgcc aacgatcaga tggcgctggg cgcaatgcgc gccattaccg 4621 agtccgggct gcgcgttggt gcggatatct cggtagtggg atacgacgat accgaagata 4681 gctcatgtta tatcccgccg ttaaccacca tcaaacagga ttttcgcctg ctggggcaaa 4741 ccagcgtgga ccgcttgctg caactctctc agggccaggc ggtgaagggc aatcagctgt 4801 tgccagtctc actggtgaaa agaaaaacca ccctggcgcc caatacgcaa accgcctctc 4861 cccgcgcgtt ggccgattca ttaatgcagc tggcacgaca ggtttcccga ctggaaagcg 4921 ggcagtgata atccaggagg aaaaaaatgt ccagattaga taaaagtaaa gtgattaaca 4981 gcgcattaga gctgcttaat gaggtcggaa tcgaaggttt aacaacccgt aaactcgccc 5041 agaagctagg tgtagagcag cctacattgt attggcatgt aaaaaataag cgggctttgc 5101 tcgacgcctt agccattgag atgttagata ggcaccatac tcacttttgc cctttagaag 5161 gggaaagctg gcaagatttt ttacgtaata acgctaaaag ttttagatgt gctttactaa 5221 gtcatcgcga tggagcaaaa gtacatttag gtacacggcc tacagaaaaa cagtatgaaa 5281 ctctcgaaaa tcaattagcc tttttatgcc aacaaggttt ttcactagag aatgcattat 5341 atgcactcag cgctgtgggg cattttactt taggttgcgt attggaagat caagagcatc 5401 aagtcgctaa agaagaaagg gaaacaccta ctactgatag tatgccgcca ttattacgac 5461 aagctatcga attatttgat caccaaggtg cagagccagc cttcttattc ggccttgaat 5521 tgatcatatg cggattagaa aaacaactta aatgtgaaag tgggtcctaa taattggtaa 5581 cgaatcagac aattgacggc tcgagggagt agcatagggt ttgcagaatc cctgcttcgt 5641 ccatttgaca ggcacattat gcatcgatga taagctgtca aacatgagca gatcctctac 5701 gccggacgca tcgtggccgg catcaccggc gccacaggtg cggttgctgg cgcctatatc 5761 gccgacatca ccgatgggga agatcgggct cgccacttcg ggctcatgag caaatatttt 5821 atctgaggt //