LOCUS Exported 5726 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION LitR-L1 gate. ACCESSION . VERSION . KEYWORDS pLitR-L1 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 5726) 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 5726) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..5726 /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 2777..2826 /label=sTRSV HHRz /note="hammerhead ribozyme from the tobacco ringspot virus satellite RNA (Khvorova et al., 2003)" CDS 2876..3595 /codon_start=1 /product="enhanced YFP" /label=EYFP /note="mammalian codon-optimized" /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KFICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKLHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL EFVTAAGITLGMDELYK" primer_bind complement(2921..2942) /label=EGFP-N /note="EGFP, reverse primer" primer_bind complement(3182..3201) /label=EXFP-R /note="For distinguishing EGFP variants, reverse primer" primer_bind 3529..3550 /label=EGFP-C /note="EGFP, forward primer" CDS 3743..4825 /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(3762..3781) /label=LacI-R /note="LacI, reverse primer" CDS 4844..5467 /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(4917..4936) /label=Tet-R /note="Tetracycline resistance gene, reverse primer" primer_bind complement(5650..5669) /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 tagtcataag tctgggctaa gcccactgat gagtcgctga aatgcgacga 1981 aacttatgac ctctacaaat aattttgttt aagtcctatg gactttttca tacaggagaa 2041 ccctcgatgg ataccattca gaaacgtccg cgtacccgtc tgagtccgga aaaacgtaaa 2101 gaacagctgc tggatattgc cattgaagtt tttagccagc gtggtattgg tcgtggtggt 2161 catgcagata ttgcagaaat tgcacaggtt agcgttgcaa ccgtgtttaa ctattttccg 2221 acccgtgaag atctggttga tgatgttctg aacaaagtgg aaaacgagtt tcaccagttc 2281 atcaataaca gcattagcct ggatctggat gttcgtagca atctgaatac cctgctgctg 2341 aacattattg atagcgttca gaccggcaac aaatggatta aagtttggtt tgaatggtca 2401 accagcaccc gtgatgaagt ttggcctctg tttctgagca cccatagcaa taccaatcag 2461 gtgatcaaaa ccatgtttga agagggtatt gaacgcaatg aagtgtgcaa tgatcataca 2521 ccggaaaatc tgaccaaaat gctgcatggt atttgctata gcgtgtttat tcaggccaat 2581 cgtaatagca gcagcgaaga aatggaagaa accgcaaatt gctttctgaa tatgctgtgc 2641 atctacaaat aactcggtac caaattccag aaaagacacc cgaaagggtg ttttttcgtt 2701 ttggtcctac tcgagcgtag agcttagatt cgttaccaat tgacaaattt ataaattgtc 2761 agtacagtcc tagcagctgt caccggatgt gctttccggt ctgatgagtc cgtgaggacg 2821 aaacagcctc tacaaataat tttgtttaat actagagaaa gaggggaaat actagatggt 2881 gagcaagggc gaggagctgt tcaccggggt ggtgcccatc ctggtcgagc tggacggcga 2941 cgtaaacggc cacaagttca gcgtgtccgg cgagggcgag ggcgatgcca cctacggcaa 3001 gctgaccctg aagttcatct gcaccacagg caagctgccc gtgccctggc ccaccctcgt 3061 gaccaccttc ggctacggcc tgcaatgctt cgcccgctac cccgaccaca tgaagctgca 3121 cgacttcttc aagtccgcca tgcccgaagg ctacgtccag gagcgcacca tcttcttcaa 3181 ggacgacggc aactacaaga cccgcgccga ggtgaagttc gagggcgaca ccctggtgaa 3241 ccgcatcgag ctgaagggca tcgacttcaa ggaggacggc aacatcctgg ggcacaagct 3301 ggagtacaac tacaacagcc acaacgtcta tatcatggcc gacaagcaga agaacggcat 3361 caaggtgaac ttcaagatcc gccacaacat cgaggacggc agcgtgcagc tcgccgacca 3421 ctaccagcag aacaccccaa tcggcgacgg ccccgtgctg ctgcccgaca accactacct 3481 tagctaccag tccgccctga gcaaagaccc caacgagaag cgcgatcaca tggtcctgct 3541 ggagttcgtg accgccgccg ggatcactct cggcatggac gagctgtaca agtaactcgg 3601 taccaaattc cagaaaagag gcctcccgaa aggggggcct tttttcgttt tggtccaatg 3661 gcggcgcgcc atcgaatggc gcaaaacctt tcgcggtatg gcatgatagc gcccggaaga 3721 gagtcaattc agggtggtga atatgaaacc agtaacgtta tacgatgtcg cagagtatgc 3781 cggtgtctct tatcagaccg tttcccgcgt ggtgaaccag gccagccacg tttctgcgaa 3841 aacgcgggaa aaagtggaag cggcgatggc ggagctgaat tacattccca accgcgtggc 3901 acaacaactg gcgggcaaac agtcgttgct gattggcgtt gccacctcca gtctggccct 3961 gcacgcgccg tcgcaaattg tcgcggcgat taaatctcgc gccgatcaac tgggtgccag 4021 cgtggtggtg tcgatggtag aacgaagcgg cgtcgaagcc tgtaaagcgg cggtgcacaa 4081 tcttctcgcg caacgcgtca gtgggctgat cattaactat ccgctggatg accaggatgc 4141 cattgctgtg gaagctgcct gcactaatgt tccggcgtta tttcttgatg tctctgacca 4201 gacacccatc aacagtatta ttttctccca tgaggacggt acgcgactgg gcgtggagca 4261 tctggtcgca ttgggtcacc agcaaatcgc gctgttagcg ggcccattaa gttctgtctc 4321 ggcgcgtctg cgtctggctg gctggcataa atatctcact cgcaatcaaa ttcagccgat 4381 agcggaacgg gaaggcgact ggagtgccat gtccggtttt caacaaacca tgcaaatgct 4441 gaatgagggc atcgttccca ctgcgatgct ggttgccaac gatcagatgg cgctgggcgc 4501 aatgcgcgcc attaccgagt ccgggctgcg cgttggtgcg gatatctcgg tagtgggata 4561 cgacgatacc gaagatagct catgttatat cccgccgtta accaccatca aacaggattt 4621 tcgcctgctg gggcaaacca gcgtggaccg cttgctgcaa ctctctcagg gccaggcggt 4681 gaagggcaat cagctgttgc cagtctcact ggtgaaaaga aaaaccaccc tggcgcccaa 4741 tacgcaaacc gcctctcccc gcgcgttggc cgattcatta atgcagctgg cacgacaggt 4801 ttcccgactg gaaagcgggc agtgataatc caggaggaaa aaaatgtcca gattagataa 4861 aagtaaagtg attaacagcg cattagagct gcttaatgag gtcggaatcg aaggtttaac 4921 aacccgtaaa ctcgcccaga agctaggtgt agagcagcct acattgtatt ggcatgtaaa 4981 aaataagcgg gctttgctcg acgccttagc cattgagatg ttagataggc accatactca 5041 cttttgccct ttagaagggg aaagctggca agatttttta cgtaataacg ctaaaagttt 5101 tagatgtgct ttactaagtc atcgcgatgg agcaaaagta catttaggta cacggcctac 5161 agaaaaacag tatgaaactc tcgaaaatca attagccttt ttatgccaac aaggtttttc 5221 actagagaat gcattatatg cactcagcgc tgtggggcat tttactttag gttgcgtatt 5281 ggaagatcaa gagcatcaag tcgctaaaga agaaagggaa acacctacta ctgatagtat 5341 gccgccatta ttacgacaag ctatcgaatt atttgatcac caaggtgcag agccagcctt 5401 cttattcggc cttgaattga tcatatgcgg attagaaaaa caacttaaat gtgaaagtgg 5461 gtcctaataa ttggtaacga atcagacaat tgacggctcg agggagtagc atagggtttg 5521 cagaatccct gcttcgtcca tttgacaggc acattatgca tcgatgataa gctgtcaaac 5581 atgagcagat cctctacgcc ggacgcatcg tggccggcat caccggcgcc acaggtgcgg 5641 ttgctggcgc ctatatcgcc gacatcaccg atggggaaga tcgggctcgc cacttcgggc 5701 tcatgagcaa atattttatc tgaggt //