LOCUS       AY048742                2804 bp    DNA     circular SYN 11-SEP-2001
DEFINITION  Template plasmid pKD3, complete sequence.
ACCESSION   AY048742
VERSION     AY048742.1  GI:15554329
KEYWORDS    .
SOURCE      Template plasmid pKD3
  ORGANISM  Template plasmid pKD3
            other sequences; artificial sequences; vectors.
REFERENCE   1  (bases 1 to 2804)
  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 2804)
  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..2804
                     /organism="Template plasmid pKD3"
                     /mol_type="genomic DNA"
                     /db_xref="taxon:170491"
     misc_feature    31..50
                     /note="priming site 1"
     misc_feature    32..51
                     /note="priming site 3"
     misc_feature    complement(51..84)
                     /note="distal 34-nt of of natural FRT site"
     CDS             complement(134..772)
                     /note="Tn9"
                     /codon_start=1
                     /transl_table=11
                     /product="chloramphenicol acetyl transferase"
                     /protein_id="AAL02033.1"
                     /db_xref="GI:15554330"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAF
                     LKTVKKNKHKFYPAFIHILARLMNAHPELRMAMKDGELVIWDSVHPCYTVFHEQTETF
                     SSLWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNV
                     ANMDNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRCLMNTTVLR"
     misc_feature    245..264
                     /note="common priming site c1"
     misc_feature    955..974
                     /note="common priming site c2"
     misc_feature    complement(981..1022)
                     /note="distal 42-nt of natural FRT site"
     misc_feature    1044..1063
                     /note="priming site 2"
     rep_origin      1047..1446
                     /note="R6K gamma replication origin"
     CDS             complement(1516..2376)
                     /note="bla; ampicillin resistance"
                     /codon_start=1
                     /transl_table=11
                     /product="beta-lactamase"
                     /protein_id="AAL02034.1"
                     /db_xref="GI:15554331"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGY
                     IELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVE
                     YSPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRL
                     DRWEPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPL
                     LRSALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIA
                     EIGASLIKHW"
     terminator      2523..2782
                     /note="rrnB transcription terminator"
ORIGIN      
        1 agattgcagc attacacgtc ttgagcgatt gtgtaggctg gagctgcttc gaagttccta
       61 tactttctag agaataggaa cttcggaata ggaacttcat ttaaatggcg cgccttacgc
      121 cccgccctgc cactcatcgc agtactgttg tattcattaa gcatctgccg acatggaagc
      181 catcacaaac ggcatgatga acctgaatcg ccagcggcat cagcaccttg tcgccttgcg
      241 tataatattt gcccatggtg aaaacggggg cgaagaagtt gtccatattg gccacgttta
      301 aatcaaaact ggtgaaactc acccagggat tggctgagac gaaaaacata ttctcaataa
      361 accctttagg gaaataggcc aggttttcac cgtaacacgc cacatcttgc gaatatatgt
      421 gtagaaactg ccggaaatcg tcgtggtatt cactccagag cgatgaaaac gtttcagttt
      481 gctcatggaa aacggtgtaa caagggtgaa cactatccca tatcaccagc tcaccgtctt
      541 tcattgccat acgtaattcc ggatgagcat tcatcaggcg ggcaagaatg tgaataaagg
      601 ccggataaaa cttgtgctta tttttcttta cggtctttaa aaaggccgta atatccagct
      661 gaacggtctg gttataggta cattgagcaa ctgactgaaa tgcctcaaaa tgttctttac
      721 gatgccattg ggatatatca acggtggtat atccagtgat ttttttctcc attttagctt
      781 ccttagctcc tgaaaatctc gacaactcaa aaaatacgcc cggtagtgat cttatttcat
      841 tatggtgaaa gttggaacct cttacgtgcc gatcaacgtc tcattttcgc caaaagttgg
      901 cccagggctt cccggtatca acagggacac caggatttat ttattctgcg aagtgatctt
      961 ccgtcacagg taggcgcgcc gaagttccta tactttctag agaataggaa cttcggaata
     1021 ggaactaagg aggatattca tatggaccat ggctaattcc catgtcagcc gttaagtgtt
     1081 cctgtgtcac tgaaaattgc tttgagaggc tctaagggct tctcagtgcg ttacatccct
     1141 ggcttgttgt ccacaaccgt taaaccttaa aagctttaaa agccttatat attctttttt
     1201 ttcttataaa acttaaaacc ttagaggcta tttaagttgc tgatttatat taattttatt
     1261 gttcaaacat gagagcttag tacgtgaaac atgagagctt agtacgttag ccatgagagc
     1321 ttagtacgtt agccatgagg gtttagttcg ttaaacatga gagcttagta cgttaaacat
     1381 gagagcttag tacgtgaaac atgagagctt agtacgtact atcaacaggt tgaactgcgg
     1441 atcttgcggc cgcaaaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta
     1501 aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct
     1561 atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg
     1621 cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga
     1681 tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt
     1741 atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt
     1801 taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt
     1861 tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat
     1921 gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc
     1981 cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc
     2041 cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat
     2101 gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag
     2161 aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt
     2221 accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc
     2281 ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa
     2341 gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg
     2401 aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa
     2461 taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgcat cgatggcccc
     2521 ccgatggtag tgtggggtct ccccatgcga gagtagggaa ctgccaggca tcaaataaaa
     2581 cgaaaggctc agtcgaaaga ctgggccttt cgttttatct gttgtttgtc ggtgaacgct
     2641 ctcctgagta ggacaaatcc gccgggagcg gatttgaacg ttgcgaagca acggcccgga
     2701 gggtggcggg caggacgccc gccataaact gccaggcatc aaattaagca gaaggccatc
     2761 ctgacggatg gcctttttgc gtggccagtg ccaagcttgc atgc
//

