LOCUS Exported 4355 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Self replication Level T acceptor vector with LacZ compatible blue/white screening. Modified SEVA vector, RK2 ori vector. . ACCESSION . VERSION . KEYWORDS pCAT.011 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 4355) AUTHORS Vasudevan R, Gale GAR, Schiavon AA, Puzorjov A, Malin J, Gillespie MD, Vavitsas K, Zulkower V, Wang B, Howe CJ, Lea-Smith DJ, McCormick AJ TITLE CyanoGate: A Modular Cloning Suite for Engineering Cyanobacteria Based on the Plant MoClo Syntax. JOURNAL Plant Physiol. 2019 May;180(1):39-55. doi: 10.1104/pp.18.01401. Epub 2019 Feb 28. PUBMED 30819783 REFERENCE 2 (bases 1 to 4355) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..4355 /organism="synthetic DNA construct" /mol_type="other DNA" protein_bind 121..142 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." promoter 157..187 /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 195..211 /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)." primer_bind 200..222 /label=M13/pUC Reverse /note="In lacZ gene" primer_bind 219..235 /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind 219..235 /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" CDS 231..554 /codon_start=1 /gene="lacZ fragment" /product="LacZ-alpha fragment of beta-galactosidase" /label=lacZ-alpha /translation="MTMITPSLHACRSTLEDPRVPSSNSLAVVLQRRDWENPGVTQLNR LAAHPPFASWRNSEEARTDRPSQQLRSLNGEWRLMRYFLLTHLCGISHRIWCTLSTICS DAA" misc_feature 248..304 /label=MCS /note="pUC18/19 multiple cloning site" primer_bind complement(305..322) /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind complement(305..321) /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" primer_bind complement(314..336) /label=M13/pUC Forward /note="In lacZ gene" primer_bind complement(530..549) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" terminator 612..706 /label=lambda t0 terminator /note="transcription terminator from phage lambda" CDS 942..1733 /codon_start=1 /gene="aadA" /product="aminoglycoside adenylyltransferase (Murphy, 1985)" /label=SmR /note="confers resistance to spectinomycin and streptomycin" /translation="MREAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK" oriT 1879..1987 /note="incP origin of transfer" CDS complement(2048..3196) /codon_start=1 /product="trans-acting replication protein that binds to and activates oriV" /label=trfA /translation="MNRTFDRKAYRQELIDAGFSAEDAETIASRTVMRAPRETFQSVGS IVQQATAKIERDSVQLAPPALPAPSAAVERSRRLEQEAAGLAKSMTIDTRGTMTTKKRK TAGEDLAKQVSEAKQAALLKHTKQQIKEMQLSLFDIAPWPDTMRAMPNDTARSALFTTR NKKIPREALQNKVIFHVNKDVKITYTGVELRADDDELVWQQVLEYAKRTPIGEPITFTF YELCQDLGWSINGRYYTKAEECLSRLQATAMGFTSDRVGHLESVSLLHRFRVLDRGKKT SRCQVLIDEEIVVLFAGDHYTKFIWEKYRKLSPTARRMFDYFSSHREPYPLKLETFRLM CGSDSTRVKKWREQVGEACEELRGSGLVEHAWVNDDLVHCKR" rep_origin 3589..4120 /label=oriV /note="incP origin of replication" terminator 4247..4333 /gene="Escherichia coli rrnB" /label=rrnB T1 terminator /note="transcription terminator T1 from the E. coli rrnB gene" ORIGIN 1 ttaagaagcg gaagagcgcc caatacgcaa accgcctctc cccgcgcgtt ggccgattca 61 ttaatgcagc tggcacgaca ggtttcccga ctggaaagcg ggcagtgagc gcaacgcaat 121 taatgtgagt tagctcactc attaggcacc ccaggcttta cactttatgc ttccggctcg 181 tatgttgtgt ggaattgtga gcggataaca atttcacaca ggaaacagct atgaccatga 241 ttacgccaag cttgcatgcc tgcaggtcga ctctagagga tccccgggta ccgagctcga 301 attcactggc cgtcgtttta caacgtcgtg actgggaaaa ccctggcgtt acccaactta 361 atcgccttgc agcacatccc cctttcgcca gctggcgtaa tagcgaagag gcccgcaccg 421 atcgcccttc ccaacagttg cgcagcctga atggcgaatg gcgcctgatg cggtattttc 481 tccttacgca tctgtgcggt atttcacacc gcatatggtg cactctcagt acaatctgct 541 ctgatgccgc atagttaagc cagccccgac acccgccaac acccgctgaa ctagtcttgg 601 aagactcggg agactcctgt tgatagatcc agtaatgacc tcagaactcc atctggattt 661 gttcagaacg ctcggttgcc gccgggcgtt ttttattggt gagaatccag gggtccccaa 721 taattacgat ttacgtattt aaatgaacct tgaccgaacg cagcggtggt aacggcgcag 781 tggcggtttt catggcttgt tatgactgtt tttttggggt acagtctatg cctcgggcat 841 ccaagcagca agcgcgttac gccgtgggtc gatgtttgat gttatggagc agcaacgatg 901 ttacgcagca gggcagtcgc cctaaaacaa agttaaacat catgagggaa gcggtgatcg 961 ccgaagtatc gactcaacta tcagaggtag ttggcgtcat cgagcgccat ctcgaaccga 1021 cgttgctggc cgtacatttg tacggctccg cagtggatgg cggcctgaag ccacacagtg 1081 atattgattt gctggttacg gtgaccgtaa ggcttgatga aacaacgcgg cgagctttga 1141 tcaacgacct tttggaaact tcggcttccc ctggagagag cgagattctc cgcgctgtag 1201 aagtcaccat tgttgtgcac gacgacatca ttccgtggcg ttatccagct aagcgcgaac 1261 tgcaatttgg agaatggcag cgcaatgaca ttcttgcagg tatcttcgag ccagccacga 1321 tcgacattga tctggctatc ttgctgacaa aagcaagaga acatagcgtt gccttggtag 1381 gtccagcggc ggaggaactc tttgatccgg ttcctgaaca ggatctattt gaggcgctaa 1441 atgaaacctt aacgctatgg aactcgccgc ccgactgggc tggcgatgag cgaaatgtag 1501 tgcttacgtt gtcccgcatt tggtacagcg cagtaaccgg caaaatcgcg ccgaaggatg 1561 tcgctgccga ctgggcaatg gagcgcctgc cggcccagta tcagcccgtc atacttgaag 1621 ctagacaggc ttatcttgga caagaagaag atcgcttggc ctcgcgcgca gatcagttgg 1681 aagaatttgt ccactacgtg aaaggcgaga tcaccaaggt agtcggcaaa taagacaatt 1741 gtccttttcc gctgcataac cctgcttcgg ggtcattata gcgatttttt cggtatatcc 1801 atcctttttc gcacgatata caggattttg ccaaagggtt cgtgtagact ttccttggtg 1861 tatccaacgg cgtcagccgg gcaggatagg tgaagtaggc ccacccgcga gcgggtgttc 1921 cttcttcact gtcccttatt cgcacctggc ggtgctcaac gggaatcctg ctctgcgagg 1981 ctggccgtag gccggccgcg atgcaggtgg ctgctgaacc cccagccgga actgacccca 2041 caaggcccta gcgtttgcaa tgcaccaggt catcattgac ccaggcgtgt tccaccaggc 2101 cgctgcctcg caactcttcg caggcttcgc cgacctgctc gcgccacttc ttcacgcggg 2161 tggaatccga tccgcacatg aggcggaagg tttccagctt gagcgggtac ggctcccggt 2221 gcgagctgaa atagtcgaac atccgtcggg ccgtcggcga cagcttgcgg tacttctccc 2281 atatgaattt cgtgtagtgg tcgccagcaa acagcacgac gatttcctcg tcgatcagga 2341 cctggcaacg ggacgttttc ttgccacggt ccaggacgcg gaagcggtgc agcagcgaca 2401 ccgattccag gtgcccaacg cggtcggacg tgaagcccat cgccgtcgcc tgtaggcgcg 2461 acaggcattc ctcggccttc gtgtaatacc ggccattgat cgaccagccc aggtcctggc 2521 aaagctcgta gaacgtgaag gtgatcggct cgccgatagg ggtgcgcttc gcgtactcca 2581 acacctgctg ccacaccagt tcgtcatcgt cggcccgcag ctcgacgccg gtgtaggtga 2641 tcttcacgtc cttgttgacg tggaaaatga ccttgttttg cagcgcctcg cgcgggattt 2701 tcttgttgcg cgtggtgaac agggcagagc gggccgtgtc gtttggcatc gctcgcatcg 2761 tgtccggcca cggcgcaata tcgaacaagg aaagctgcat ttccttgatc tgctgcttcg 2821 tgtgtttcag caacgcggcc tgcttggctt cgctgacctg ttttgccagg tcctcgccgg 2881 cggtttttcg cttcttggtc gtcatagttc ctcgcgtgtc gatggtcatc gacttcgcca 2941 aacctgccgc ctcctgttcg agacgacgcg aacgctccac ggcggccgat ggcgcgggca 3001 gggcaggggg agccagttgc acgctgtcgc gctcgatctt ggccgtagct tgctggacta 3061 tcgagccgac ggactggaag gtttcgcggg gcgcacgcat gacggtgcgg cttgcgatgg 3121 tttcggcatc ctcggcggaa aaccccgcgt cgatcagttc ttgcctgtat gccttccggt 3181 caaacgtccg attcattcac cctccttgcg ggattgcccc ggaattaatt ccccggatcg 3241 atccgtcgat cttgatcccc tgcgccatca gatccttggc ggcaagaaag ccatccagtt 3301 tactttgcag ggcttcccaa ccttaccaga gggcgcccca gctggcaatt ccggttcgct 3361 tgctgtccat aaaaccgccc agtctagcta tcgccatgta agcccactgc aagctacctg 3421 ctttctcttt gcgcttgcgt tttcccttgt ccagatagcc cagtagctga cattcatccg 3481 gggtcagcac cgtttctgcg gactggcttt ctacgtggct gccatttttg gggtgaggcc 3541 gttcgcggcc gaggggcgca gcccctgggg ggatgggagg cccgcgttag cgggccggga 3601 gggttcgaga agggggggca ccccccttcg gcgtgcgcgg tcacgcgcac agggcgcagc 3661 cctggttaaa aacaaggttt ataaatattg gtttaaaagc aggttaaaag acaggttagc 3721 ggtggccgaa aaacgggcgg aaacccttgc aaatgctgga ttttctgcct gtggacagcc 3781 cctcaaatgt caataggtgc gcccctcatc tgtcagcact ctgcccctca agtgtcaagg 3841 atcgcgcccc tcatctgtca gtagtcgcgc ccctcaagtg tcaataccgc agggcactta 3901 tccccaggct tgtccacatc atctgtggga aactcgcgta aaatcaggcg ttttcgccga 3961 tttgcgaggc tggccagctc cacgtcgccg gccgaaatcg agcctgcccc tcatctgtca 4021 acgccgcgcc gggtgagtcg gcccctcaag tgtcaacgtc cgcccctcat ctgtcagtga 4081 gggccaagtt ttccgcgagg tatccacaac gccggcggcc ctacatggct ctgctgtagt 4141 gagtgggttg cgctccggca gcggtcctga tcccccgcag aaaaaaagga tctcaagaag 4201 atcctttgat cttttctacg gcgcgcccag ctgtctaggg cggcggattt gtcctactca 4261 ggagagcgtt caccgacaaa caacagataa aacgaaaggc ccagtctttc gactgagcct 4321 ttcgttttat ttgatgcctt gccgagtctt cttaa //