LOCUS AY048743 3267 bp DNA circular SYN 11-SEP-2001 DEFINITION Template plasmid pKD4, complete sequence. ACCESSION AY048743 VERSION AY048743.1 GI:15554332 KEYWORDS . SOURCE Template plasmid pKD4 ORGANISM Template plasmid pKD4 other sequences; artificial sequences; vectors. REFERENCE 1 (bases 1 to 3267) AUTHORS Datsenko,K.A. and Wanner,B.L. TITLE One-step inactivation of chromosomal genes in Escherichia coli K-12 using PCR products JOURNAL Proc. Natl. Acad. Sci. U.S.A. 97 (12), 6640-6645 (2000) PUBMED 10829079 REFERENCE 2 (bases 1 to 3267) AUTHORS Datsenko,K.A. and Wanner,B.L. TITLE Direct Submission JOURNAL Submitted (29-JUL-2001) Biological Sciences, Purdue University, Lilly Hall of Life Sciences, West Lafayette, IN 47907, USA FEATURES Location/Qualifiers source 1..3267 /organism="Template plasmid pKD4" /mol_type="genomic DNA" /db_xref="taxon:170492" misc_feature 31..50 /note="priming site 1" misc_feature 32..51 /note="priming site 3" misc_feature complement(51..98) /note="natural FRT site" CDS 459..1253 /note="kanamycin resistance" /codon_start=1 /transl_table=11 /product="Tn5 neomycin phosphotransferase" /protein_id="AAL02035.1" /db_xref="GI:15554333" /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGR PVLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDL LSSHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDE EHQGLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRY QDIALATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF" misc_feature 529..548 /note="common priming site k1" misc_feature 620..639 /note="common priming site k2" misc_feature 1090..1109 /note="common priming site kt" misc_feature complement(1444..1489) /note="FRT site" misc_feature 1507..1526 /note="priming site 2" rep_origin 1523..1909 /note="R6K gamma replication origin" CDS complement(1979..2839) /note="bla; ampicillin resistance" /codon_start=1 /transl_table=11 /product="beta-lactamase" /protein_id="AAL02036.1" /db_xref="GI:15554334" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGY IELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVE YSPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRL DRWEPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPL LRSALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIA EIGASLIKHW" terminator 2986..3245 /note="rrnB transcription terminator" ORIGIN 1 agattgcagc attacacgtc ttgagcgatt gtgtaggctg gagctgcttc gaagttccta 61 tactttctag agaataggaa cttcggaata ggaacttcaa gatcccctca cgctgccgca 121 agcactcagg gcgcaagggc tgctaaagga agcggaacac gtagaaagcc agtccgcaga 181 aacggtgctg accccggatg aatgtcagct actgggctat ctggacaagg gaaaacgcaa 241 gcgcaaagag aaagcaggta gcttgcagtg ggcttacatg gcgatagcta gactgggcgg 301 ttttatggac agcaagcgaa ccggaattgc cagctggggc gccctctggt aaggttggga 361 agccctgcaa agtaaactgg atggctttct tgccgccaag gatctgatgg cgcaggggat 421 caagatctga tcaagagaca ggatgaggat cgtttcgcat gattgaacaa gatggattgc 481 acgcaggttc tccggccgct tgggtggaga ggctattcgg ctatgactgg gcacaacaga 541 caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt 601 ttgtcaagac cgacctgtcc ggtgccctga atgaactgca ggacgaggca gcgcggctat 661 cgtggctggc cacgacgggc gttccttgcg cagctgtgct cgacgttgtc actgaagcgg 721 gaagggactg gctgctattg ggcgaagtgc cggggcagga tctcctgtca tctcaccttg 781 ctcctgccga gaaagtatcc atcatggctg atgcaatgcg gcggctgcat acgcttgatc 841 cggctacctg cccattcgac caccaagcga aacatcgcat cgagcgagca cgtactcgga 901 tggaagccgg tcttgtcgat caggatgatc tggacgaaga gcatcagggg ctcgcgccag 961 ccgaactgtt cgccaggctc aaggcgcgca tgcccgacgg cgaggatctc gtcgtgaccc 1021 atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct ggattcatcg 1081 actgtggccg gctgggtgtg gcggaccgct atcaggacat agcgttggct acccgtgata 1141 ttgctgaaga gcttggcggc gaatgggctg accgcttcct cgtgctttac ggtatcgccg 1201 ctcccgattc gcagcgcatc gccttctatc gccttcttga cgagttcttc tgagcgggac 1261 tctggggttc gaaatgaccg accaagcgac gcccaacctg ccatcacgag atttcgattc 1321 caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg ccggctggat 1381 gatcctccag cgcggggatc tcatgctgga gttcttcgcc caccccagct tcaaaagcgc 1441 tctgaagttc ctatactttc tagagaatag gaacttcgga ataggaacta aggaggatat 1501 tcatatggac catggctaat tcccatgtca gccgttaagt gttcctgtgt cactgaaaat 1561 tgctttgaga ggctctaagg gcttctcagt gcgttacatc cctggcttgt tgtccacaac 1621 cgttaaacct taaaagcttt aaaagcctta tatattcttt tttttcttat aaaacttaaa 1681 accttagagg ctatttaagt tgctgattta tattaatttt attgttcaaa catgagagct 1741 tagtacgtga aacatgagag cttagtacgt tagccatgag agcttagtac gttagccatg 1801 agggtttagt tcgttaaaca tgagagctta gtacgttaaa catgagagct tagtacgtga 1861 aacatgagag cttagtacgt actatcaaca ggttgaactg cggatcttgc ggccgcaaaa 1921 attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt 1981 accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag 2041 ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca 2101 gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc 2161 agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt 2221 ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg 2281 ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca 2341 gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg 2401 ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca 2461 tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg 2521 tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct 2581 cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca 2641 tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca 2701 gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg 2761 tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac 2821 ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt 2881 attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc 2941 cgcgcacatt tccccgaaaa gtgccacctg catcgatggc cccccgatgg tagtgtgggg 3001 tctccccatg cgagagtagg gaactgccag gcatcaaata aaacgaaagg ctcagtcgaa 3061 agactgggcc tttcgtttta tctgttgttt gtcggtgaac gctctcctga gtaggacaaa 3121 tccgccggga gcggatttga acgttgcgaa gcaacggccc ggagggtggc gggcaggacg 3181 cccgccataa actgccaggc atcaaattaa gcagaaggcc atcctgacgg atggcctttt 3241 tgcgtggcca gtgccaagct tgcatgc //