LOCUS Exported 4968 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS pICH47742 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 4968) AUTHORS Weber E, Engler C, Gruetzner R, Werner S, Marillonnet S TITLE A modular cloning system for standardized assembly of multigene constructs. JOURNAL PLoS One. 2011 Feb 18;6(2):e16765. doi: 10.1371/journal.pone.0016765. PUBMED 21364738 REFERENCE 2 (bases 1 to 4968) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..4968 /organism="synthetic DNA construct" /mol_type="other DNA" misc_feature 76..100 /label=LB T-DNA repeat /note="left border repeat from nopaline C58 T-DNA" rep_origin 192..823 /label=oriV /note="incP origin of replication" promoter 910..1014 /gene="bla" /label=AmpR promoter CDS 1015..1875 /codon_start=1 /gene="bla" /product="beta-lactamase" /label=AmpR /note="confers resistance to ampicillin, carbenicillin, and related antibiotics" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS LIKHW" primer_bind complement(1233..1252) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 2046..2634 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 2535..2554 /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS 2970..4118 /codon_start=1 /product="trans-acting replication protein that binds to and activates oriV" /label=trfA /translation="MNRTFDRKAYRQELIDAGFSAEDAETIASRTVMRAPRETFQSVGS MVQQATAKIERDSVQLAPPALPAPSAAVERSRRLEQEAAGLAKSMTIDTRGTMTTKKRK TAGEDLAKQVSEAKQAALLKHTKQQIKEMQLSLFDIAPWPDTMRAMPNDTARSALFTTR NKKIPREALQNKVIFHVNKDVKITYTGVELRADDDELVWQQVLEYAKRTPIGEPITFTF YELCQDLGWSINGRYYTKAEECLSRLQATAMGFTSDRVGHLESVSLLHRFRVLDRGKKT SRCQVLIDEEIVVLFAGDHYTKFIWEKYRKLSPTARRMFDYFSSHREPYPLKLETFRLM CGSDSTRVKKWREQVGEACEELRGSGLVEHAWVNDDLVHCKR" misc_feature 4274..4298 /label=RB T-DNA repeat /note="right border repeat from nopaline C58 T-DNA" CDS complement(4440..4763) /codon_start=1 /gene="lacZ fragment" /product="LacZ-alpha fragment of beta-galactosidase" /label=lacZ-alpha /translation="MTMITPSLHACRSTLEDPRVPSSNSLAVVLQRRDWENPGVTQLNR LAAHPPFASWRNSEEARTDRPSQQLRSLNGEWRLMRYFLLTHLCGISHRIWCTLSTICS DAA" primer_bind 4445..4464 /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind 4658..4680 /label=M13/pUC Forward /note="In lacZ gene" primer_bind 4672..4689 /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind 4673..4689 /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" misc_feature 4690..4746 /label=MCS /note="pUC18/19 multiple cloning site" primer_bind complement(4759..4775) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(4759..4775) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(4772..4794) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 4783..4799 /label=lac operator /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)." promoter complement(4807..4837) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 4852..4873 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." ORIGIN 1 gtggggccca ctgcatccac cccagtacat taaaaacgtc cgcaatgtgt tattaagttg 61 tctaagcgtc aatttgttta caccacaata tatcctgcca ccagccagcc aacagctccc 121 cgaccggcag ctcggcacaa aatcaccact cgatacaggc agcccatcag tcagatcagg 181 atctcctttg cgacgctcac cgggctggtt gccctcgccg ctgggctggc ggccgtctat 241 ggccctgcaa acgcgccaga aacgccgtcg aagccgtgtg cgagacaccg cggccgccgg 301 cgttgtggat acctcgcgga aaacttggcc ctcactgaca gatgaggggc ggacgttgac 361 acttgagggg ccgactcacc cggcgcggcg ttgacagatg aggggcaggc tcgatttcgg 421 ccggcgacgt ggagctggcc agcctcgcaa atcggcgaaa acgcctgatt ttacgcgagt 481 ttcccacaga tgatgtggac aagcctgggg ataagtgccc tgcggtattg acacttgagg 541 ggcgcgacta ctgacagatg aggggcgcga tccttgacac ttgaggggca gagtgctgac 601 agatgagggg cgcacctatt gacatttgag gggctgtcca caggcagaaa atccagcatt 661 tgcaagggtt tccgcccgtt tttcggccac cgctaacctg tcttttaacc tgcttttaaa 721 ccaatattta taaaccttgt ttttaaccag ggctgcgccc tgtgcgcgtg accgcgcacg 781 ccgaaggggg gtgccccccc ttctcgaacc ctcccggccc gctaacgcgg gcctcccatc 841 cccccagggg ctgcgcccct cggccgcgaa cggcctcacc ccaaaaatgg cagcgctggc 901 caattcgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg 961 ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtatgagt 1021 attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt 1081 gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg 1141 ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa 1201 cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt 1261 gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag 1321 tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt 1381 gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac gatcggagga 1441 ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt 1501 tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgta 1561 gcaatggcaa caacgttgcg caaactatta actggcgaac tacttactct agcttcccgg 1621 caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc 1681 cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg ttctcgcggt 1741 atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg 1801 gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg 1861 attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat tgatttaaaa 1921 cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa 1981 atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga 2041 tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg 2101 ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact 2161 ggcttcagca gagcgcagat accaaatact gtccttctag tgtagccgta gttaggccac 2221 cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg 2281 gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg 2341 gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga 2401 acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc 2461 gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg 2521 agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc 2581 tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc 2641 agcaacgcgg cctttttacg gttcctggca gatcctagat gtggcgcaac gatgccggcg 2701 acaagcagga gcgcaccgac ttcttccgca tcaagtgttt tggctctcag gccgaggccc 2761 acggcaagta tttgggcaag gggtcgctgg tattcgtgca gggcaagatt cggaatacca 2821 agtacgagaa ggacggccag acggtctacg ggaccgactt cattgccgat aaggtggatt 2881 atctggacac caaggcacca ggcgggtcaa atcaggaata agggcacatt gccccggcgt 2941 gagtcggggc aatcccgcaa ggagggtgaa tgaatcggac gtttgaccgg aaggcataca 3001 ggcaagaact gatcgacgcg gggttttccg ccgaggatgc cgaaaccatc gcaagccgca 3061 ccgtcatgcg tgcgccccgc gaaaccttcc agtccgtcgg ctcgatggtc cagcaagcta 3121 cggccaagat cgagcgcgac agcgtgcaac tggctccccc tgccctgccc gcgccatcgg 3181 ccgccgtgga gcgttcgcgt cgtcttgaac aggaggcggc aggtttggcg aagtcgatga 3241 ccatcgacac gcgaggaact atgacgacca agaagcgaaa aaccgccggc gaggacctgg 3301 caaaacaggt cagcgaggcc aagcaggccg cgttgctgaa acacacgaag cagcagatca 3361 aggaaatgca gctttccttg ttcgatattg cgccgtggcc ggacacgatg cgagcgatgc 3421 caaacgacac ggcccgctct gccctgttca ccacgcgcaa caagaaaatc ccgcgcgagg 3481 cgctgcaaaa caaggtcatt ttccacgtca acaaggacgt gaagatcacc tacaccggcg 3541 tcgagctgcg ggccgacgat gacgaactgg tgtggcagca ggtgttggag tacgcgaagc 3601 gcacccctat cggcgagccg atcaccttca cgttctacga gctttgccag gacctgggct 3661 ggtcgatcaa tggccggtat tacacgaagg ccgaggaatg cctgtcgcgc ctacaggcga 3721 cggcgatggg cttcacgtcc gaccgcgttg ggcacctgga atcggtgtcg ctgctgcacc 3781 gcttccgcgt cctggaccgt ggcaagaaaa cgtcccgttg ccaggtcctg atcgacgagg 3841 aaatcgtcgt gctgtttgct ggcgaccact acacgaaatt catatgggag aagtaccgca 3901 agctgtcgcc gacggcccga cggatgttcg actatttcag ctcgcaccgg gagccgtacc 3961 cgctcaagct ggaaaccttc cgcctcatgt gcggatcgga ttccacccgc gtgaagaagt 4021 ggcgcgagca ggtcggcgaa gcctgcgaag agttgcgagg cagcggcctg gtggaacacg 4081 cctgggtcaa tgatgacctg gtgcattgca aacgctaggg ccttgtgggg tcagttccgg 4141 ctgggggttc agcagccagc gcctgatctg gggaaccctg tggttggcat gcacatacaa 4201 atggacgaac ggataaacct tttcacgccc ttttaaatat ccgattattc taataaacgc 4261 tcttttctct taggtttacc cgccaatata tcctgtcaaa cactgatagt ttaaaccact 4321 tcgtgcagaa gacaatagta gcgtgagacc gtcacagctt gtctgtaagc ggatgccggg 4381 agcagacaag cccgtcaggg cgcgtcagcg ggtgttggcg ggtgtcgggg ctggcttaac 4441 tatgcggcat cagagcagat tgtactgaga gtgcaccata tgcggtgtga aataccgcac 4501 agatgcgtaa ggagaaaata ccgcatcagg cgccattcgc cattcaggct gcgcaactgt 4561 tgggaagggc gatcggtgcg ggcctcttcg ctattacgcc agctggcgaa agggggatgt 4621 gctgcaaggc gattaagttg ggtaacgcca gggttttccc agtcacgacg ttgtaaaacg 4681 acggccagtg aattcgagct cggtacccgg ggatcctcta gagtcgacct gcaggcatgc 4741 aagcttggcg taatcatggt catagctgtt tcctgtgtga aattgttatc cgctcacaat 4801 tccacacaac atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag 4861 ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcggcaa acctgtcgtg 4921 ccagctgcgg tctcactcca agcttgaatt cttgcttgtc ttcacaga //