LOCUS Exported 2842 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS pKD3 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 2842) AUTHORS Datsenko KA, Wanner BL TITLE One-step inactivation of chromosomal genes in Escherichia coli K-12 using PCR products. JOURNAL Proc Natl Acad Sci U S A. 2000 Jun 6;97(12):6640-5. PUBMED 10829079 REFERENCE 2 (bases 1 to 2842) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..2842 /organism="synthetic DNA construct" /mol_type="other DNA" terminator 107..193 /gene="Escherichia coli rrnB" /label=rrnB T1 terminator /note="transcription terminator T1 from the E. coli rrnB gene" terminator 285..312 /label=rrnB T2 terminator /note="transcription terminator T2 from the E. coli rrnB gene" protein_bind complement(390..437) /label=FRT /bound_moiety="FLP recombinase from the Saccharomyces cerevisiae 2u plasmid" /note="FLP-mediated recombination occurs in the 8-bp core sequence TCTAGAAA (Turan and Bode, 2011)." CDS complement(454..1113) /codon_start=1 /gene="cat" /product="chloramphenicol acetyltransferase" /label=CmR /note="confers resistance to chloramphenicol" /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL KTVKKNKHKFYPAFIHILARLMNAHPELRMAMKDGELVIWDSVHPCYTVFHEQTETFSS LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA" primer_bind 1028..1047 /label=CAT-R /note="Chloramphenicol resistance gene, reverse primer" promoter complement(1114..1216) /label=cat promoter /note="promoter of the E. coli cat gene encoding chloramphenicol acetyltransferase" protein_bind complement(1322..1355) /label=FRT (minimal) /bound_moiety="FLP recombinase from the Saccharomyces cerevisiae 2u plasmid" /note="supports FLP-mediated excision but not integration (Turan and Bode, 2011)" rep_origin 1404..1792 /label=R6K gamma ori /note="gamma replication origin from E. coli plasmid R6K; requires the R6K initiator protein pi for replication" CDS complement(1894..2754) /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 2517..2536 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(2755..17) /gene="bla" /label=AmpR promoter ORIGIN 1 caaatagggg ttccgcgcac atttccccga aaagtgccac ctgcatcgat ggcccccgat 61 ggtagtgtgg ggtctcccca tgcgagagta gggaactgcc aggcatcaaa taaaatgaaa 121 ggctcagtcg aaagactggg cctttcgttt tatctgttgt ttgtcggtga acgctctcct 181 gagtaggaca aatccgccgg gagcggattt gaacgttgcg aagcaacggc ccggagggtg 241 gcgggcagga cgcccgccat aaactgccag gcatcaaatt aagcagaagg ccatcctgac 301 ggatggcctt tttgcgtggc cagtgccaag cttgcatgca gattgcagca ttacacgtct 361 tgagcgattg tgtaggctgg agctgcttcg aagttcctat actttctaga gaataggaac 421 ttcggaatag gaacttcatt taaatggcgc gccttacgcc ccgccctgcc actcatcgca 481 gtactgttgt aattcattaa gcattctgcc gacatggaag ccatcacaaa cggcatgatg 541 aacctgaatc gccagcggca tcagcacctt gtcgccttgc gtataatatt tgcccatggt 601 gaaaacgggg gcgaagaagt tgtccatatt ggccacgttt aaatcaaaac tggtgaaact 661 cacccaggga ttggctgaga cgaaaaacat attctcaata aaccctttag ggaaataggc 721 caggttttca ccgtaacacg ccacatcttg cgaatatatg tgtagaaact gccggaaatc 781 gtcgtggtat tcactccaga gcgatgaaaa cgtttcagtt tgctcatgga aaacggtgta 841 acaagggtga acactatccc atatcaccag ctcaccgtct ttcattgcca tacgtaattc 901 cggatgagca ttcatcaggc gggcaagaat gtgaataaag gccggataaa acttgtgctt 961 atttttcttt acggtcttta aaaaggccgt aatatccagc tgaacggtct ggttataggt 1021 acattgagca actgactgaa atgcctcaaa atgttcttta cgatgccatt gggatatatc 1081 aacggtggta tatccagtga tttttttctc cattttagct tccttagctc ctgaaaatct 1141 cgacaactca aaaaatacgc ccggtagtga tcttatttca ttatggtgaa agttggaacc 1201 tcttacgtgc cgatcaacgt ctcattttcg ccaaaagttg gcccagggct tcccggtatc 1261 aacagggaca ccaggattta tttattctgc gaagtgatct tccgtcacag gtaggcgcgc 1321 cgaagttcct atactttcta gagaatagga acttcggaat aggaactaag gaggatattc 1381 atatggacca tggctaattc ccatgtcagc cgttaagtgt tcctgtgtca ctcaaaattg 1441 ctttgagagg ctctaagggc ttctcagtgc gttacatccc tggcttgttg tccacaaccg 1501 ttaaacctta aaagctttaa aagccttata tattcttttt tttcttataa aacttaaaac 1561 cttagaggct atttaagttg ctgatttata ttaattttat tgttcaaaca tgagagctta 1621 gtacgtgaaa catgagagct tagtacgtta gccatgagag cttagtacgt tagccatgag 1681 ggtttagttc gttaaacatg agagcttagt acgttaaaca tgagagctta gtacgtgaaa 1741 catgagagct tagtacgtac tatcaacagg ttgaactgct gatcttcaga tcctctacgc 1801 cggacgcatc gtggccggat cttgcggccg caaaaattaa aaatgaagtt ttaaatcaat 1861 ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc 1921 tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat 1981 aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc 2041 acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag 2101 aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag 2161 agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt 2221 ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg 2281 agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt 2341 tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc 2401 tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc 2461 attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa 2521 taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg 2581 aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc 2641 caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag 2701 gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt 2761 cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt 2821 tgaatgtatt tagaaaaata aa //