LOCUS Exported 2983 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS pZE21-GFPaav SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 2983) AUTHORS Elowitz MB, Leibler S TITLE A synthetic oscillatory network of transcriptional regulators. JOURNAL Nature. 2000 Jan 20. 403(6767):335-8. PUBMED 10659856 REFERENCE 2 (bases 1 to 2983) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..2983 /organism="synthetic DNA construct" /mol_type="other DNA" promoter 6..79 /label=PLtetO-1 promoter /note="modified phage lambda PL promoter with tet operator sites (Lutz and Bujard, 1997)" protein_bind 6..24 /gene="tetO" /label=tet operator /bound_moiety="tetracycline repressor TetR" /note="bacterial operator O2 for the tetR and tetA genes" protein_bind 31..49 /gene="tetO" /label=tet operator /bound_moiety="tetracycline repressor TetR" /note="bacterial operator O2 for the tetR and tetA genes" RBS 89..100 /note="strong bacterial ribosome binding site (Elowitz and Leibler, 2000)" CDS 109..822 /codon_start=1 /product="Aequoria victoria green fluorescent protein" /label=GFP /translation="MRKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK FICTTGKLPVPWPTLVTTFGYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE FVTAAGITHGMDELYK" primer_bind complement(139..167) /label=GFP-R /note="GFP, reverse primer. Does NOT anneal to EGFP" primer_bind 762..782 /label=GFP-F /note="GFP, forward primer. Does NOT anneal to EGFP" terminator 925..1011 /gene="Escherichia coli rrnB" /label=rrnB T1 terminator /note="transcription terminator T1 from the E. coli rrnB gene" rep_origin complement(1176..1764) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(1256..1275) /label=pBR322ori-F /note="pBR322 origin, forward primer" terminator 1852..1946 /label=lambda t0 terminator /note="transcription terminator from phage lambda" CDS complement(1977..2771) /codon_start=1 /gene="aph(3')-II (or nptII)" /product="aminoglycoside phosphotransferase from Tn5" /label=NeoR/KanR /note="confers resistance to neomycin, kanamycin, and G418 (Geneticin(R))" /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF" primer_bind complement(2088..2107) /label=Neo-F /note="Neomycin resistance gene, forward primer" primer_bind 2698..2717 /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 2950..2968 /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" ORIGIN 1 tcgagtccct atcagtgata gagattgaca tccctatcag tgatagagat actgagcaca 61 tcagcaggac gcactgaccg aattcattaa agaggagaaa ggtaccgcat gcgtaaagga 121 gaagaacttt tcactggagt tgtcccaatt cttgttgaat tagatggtga tgttaatggg 181 cacaaatttt ctgtcagtgg agagggtgaa ggtgatgcaa catacggaaa acttaccctt 241 aaatttattt gcactactgg aaaactacct gttccatggc caacacttgt cactactttc 301 ggttatggtg ttcaatgctt tgcgagatac ccagatcata tgaaacagca tgactttttc 361 aagagtgcca tgcccgaagg ttatgtacag gaaagaacta tatttttcaa agatgacggg 421 aactacaaga cacgtgctga agtcaagttt gaaggtgata cccttgttaa tagaatcgag 481 ttaaaaggta ttgattttaa agaagatgga aacattcttg gacacaaatt ggaatacaac 541 tataactcac acaatgtata catcatggca gacaaacaaa agaatggaat caaagttaac 601 ttcaaaatta gacacaacat tgaagatgga agcgttcaac tagcagacca ttatcaacaa 661 aatactccaa ttggcgatgg ccctgtcctt ttaccagaca accattacct gtccacacaa 721 tctgcccttt cgaaagatcc caacgaaaag agagaccaca tggtccttct tgagtttgta 781 acagctgctg ggattacaca tggcatggat gaactataca aaaggcctgc agcaaacgac 841 gaaaactacg ctgcatcagt ttaataagct tgatatcgaa ttcctgcagc ccgggggatc 901 ccatggtacg cgtgctagag gcatcaaata aaacgaaagg ctcagtcgaa agactgggcc 961 tttcgtttta tctgttgttt gtcggtgaac gctctcctga gtaggacaaa tccgccgccc 1021 tagacctagg gcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg 1081 ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag 1141 gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac 1201 gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga 1261 taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt 1321 accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc 1381 tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc 1441 cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta 1501 agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat 1561 gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac tagaaggaca 1621 gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct 1681 tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt 1741 acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct 1801 cagtggaacg aaaactcacg ttaagggatt ttggtcatga ctagtgcttg gattctcacc 1861 aataaaaaac gcccggcggc aaccgagcgt tctgaacaaa tccagatgga gttctgaggt 1921 cattactgga tctatcaaca ggagtccaag cgagctctcg aaccccagag tcccgctcag 1981 aagaactcgt caagaaggcg atagaaggcg atgcgctgcg aatcgggagc ggcgataccg 2041 taaagcacga ggaagcggtc agcccattcg ccgccaagct cttcagcaat atcacgggta 2101 gccaacgcta tgtcctgata gcggtccgcc acacccagcc ggccacagtc gatgaatcca 2161 gaaaagcggc cattttccac catgatattc ggcaagcagg catcgccatg ggtcacgacg 2221 agatcctcgc cgtcgggcat gcgcgccttg agcctggcga acagttcggc tggcgcgagc 2281 ccctgatgct cttcgtccag atcatcctga tcgacaagac cggcttccat ccgagtacgt 2341 gctcgctcga tgcgatgttt cgcttggtgg tcgaatgggc aggtagccgg atcaagcgta 2401 tgcagccgcc gcattgcatc agccatgatg gatactttct cggcaggagc aaggtgagat 2461 gacaggagat cctgccccgg cacttcgccc aatagcagcc agtcccttcc cgcttcagtg 2521 acaacgtcga gcacagctgc gcaaggaacg cccgtcgtgg ccagccacga tagccgcgct 2581 gcctcgtcct gcagttcatt cagggcaccg gacaggtcgg tcttgacaaa aagaaccggg 2641 cgcccctgcg ctgacagccg gaacacggcg gcatcagagc agccgattgt ctgttgtgcc 2701 cagtcatagc cgaatagcct ctccacccaa gcggccggag aacctgcgtg caatccatct 2761 tgttcaatca tgcgaaacga tcctcatcct gtctcttgat cagatcttga tcccctgcgc 2821 catcagatcc ttggcggcaa gaaagccatc cagtttactt tgcagggctt cccaacctta 2881 ccagagggcg ccccagctgg caattccgac gtctaagaaa ccattattat catgacatta 2941 acctataaaa ataggcgtat cacgaggccc tttcgtcttc acc //