LOCUS       AY048744                3434 bp    DNA     circular SYN 11-SEP-2001
DEFINITION  Template plasmid pKD13, complete sequence.
ACCESSION   AY048744
VERSION     AY048744.1  GI:15554335
KEYWORDS    .
SOURCE      Template plasmid pKD13
  ORGANISM  Template plasmid pKD13
            other sequences; artificial sequences; vectors.
REFERENCE   1  (bases 1 to 3434)
  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 3434)
  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..3434
                     /organism="Template plasmid pKD13"
                     /mol_type="genomic DNA"
                     /db_xref="taxon:170493"
     misc_feature    31..50
                     /note="priming site 1"
     misc_feature    complement(51..98)
                     /note="natural FRT site"
     CDS             complement(111..905)
                     /note="kanamycin resistance"
                     /codon_start=1
                     /transl_table=11
                     /product="Tn5 neomycin phosphotransferase"
                     /protein_id="AAL02037.1"
                     /db_xref="GI:15554336"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGR
                     PVLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDL
                     LSSHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDE
                     EHQGLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRY
                     QDIALATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     misc_feature    274..293
                     /note="common priming site kt"
     misc_feature    744..763
                     /note="common priming site k2"
     misc_feature    835..854
                     /note="common priming site k1"
     misc_feature    complement(1273..1307)
                     /note="distal 35-nt of natural FRT site"
     misc_feature    1334..1353
                     /note="priming site 4"
     terminator      1402..1644
                     /note="lambda terminator"
     rep_origin      1678..2076
                     /note="R6K gamma replication origin"
     CDS             complement(2146..3006)
                     /note="bla; ampicillin resistance"
                     /codon_start=1
                     /transl_table=11
                     /product="beta-lactamase"
                     /protein_id="AAL02038.1"
                     /db_xref="GI:15554337"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGY
                     IELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVE
                     YSPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRL
                     DRWEPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPL
                     LRSALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIA
                     EIGASLIKHW"
     terminator      3153..3412
                     /note="rrnB transcription terminator"
ORIGIN      
        1 agattgcagc attacacgtc ttgagcgatt gtgtaggctg gagctgcttc gaagttccta
       61 tactttctag agaataggaa cttcggaata ggaacttcaa gatcccctta ttagaagaac
      121 tcgtcaagaa ggcgatagaa ggcgatgcgc tgcgaatcgg gagcggcgat accgtaaagc
      181 acgaggaagc ggtcagccca ttcgccgcca agctcttcag caatatcacg ggtagccaac
      241 gctatgtcct gatagcggtc cgccacaccc agccggccac agtcgatgaa tccagaaaag
      301 cggccatttt ccaccatgat attcggcaag caggcatcgc catgggtcac gacgagatcc
      361 tcgccgtcgg gcatgcgcgc cttgagcctg gcgaacagtt cggctggcgc gagcccctga
      421 tgctcttcgt ccagatcatc ctgatcgaca agaccggctt ccatccgagt acgtgctcgc
      481 tcgatgcgat gtttcgcttg gtggtcgaat gggcaggtag ccggatcaag cgtatgcagc
      541 cgccgcattg catcagccat gatggatact ttctcggcag gagcaaggtg agatgacagg
      601 agatcctgcc ccggcacttc gcccaatagc agccagtccc ttcccgcttc agtgacaacg
      661 tcgagcacag ctgcgcaagg aacgcccgtc gtggccagcc acgatagccg cgctgcctcg
      721 tcctgcagtt cattcagggc accggacagg tcggtcttga caaaaagaac cgggcgcccc
      781 tgcgctgaca gccggaacac ggcggcatca gagcagccga ttgtctgttg tgcccagtca
      841 tagccgaata gcctctccac ccaagcggcc ggagaacctg cgtgcaatcc atcttgttca
      901 atcatgcgaa acgatcctca tcctgtctct tgatcagatc ttgatcccct gcgccatcag
      961 atccttggcg gcaagaaagc catccagttt actttgcagg gcttcccaac cttaccagag
     1021 ggcgccccag ctggcaattc cggttcgctt gctgtccata aaaccgccca gtctagctat
     1081 cgccatgtaa gcccactgca agctacctgc tttctctttg cgcttgcgtt ttcccttgtc
     1141 cagatagccc agtagctgac attcatccgg ggtcagcacc gtttctgcgg actggctttc
     1201 tacgtgttcc gcttccttta gcagcccttg cgccctgagt gcttgcggca gcgtgagctt
     1261 caaaagcgct ctgaagttcc tatactttct agagaatagg aacttcgaac tgcaggtcga
     1321 cggatccccg gaattaattc tcatgtttga cagcttatca ctgatcagtg aattaatggc
     1381 gatgacgcat cctcacgata atatccgggt aggcgcaatc actttcgtct ctactccgtt
     1441 acaaagcgag gctgggtatt tcccggcctt tctgttatcc gaaatccact gaaagcacag
     1501 cggctggctg aggagataaa taataaacga ggggctgtat gcacaaagca tcttctgttg
     1561 agttaagaac gagtatcgag atggcacata gccttgctca aattggaatc aggtttgtgc
     1621 caataccagt agaaacagac gaagaagcta gctttgcact ggattgcgag gctttgccat
     1681 ggctaattcc catgtcagcc gttaagtgtt cctgtgtcac tgaaaattgc tttgagaggc
     1741 tctaagggct tctcagtgcg ttacatccct ggcttgttgt ccacaaccgt taaaccttaa
     1801 aagctttaaa agccttatat attctttttt ttcttataaa acttaaaacc ttagaggcta
     1861 tttaagttgc tgatttatat taattttatt gttcaaacat gagagcttag tacgtgaaac
     1921 atgagagctt agtacgttag ccatgagagc ttagtacgtt agccatgagg gtttagttcg
     1981 ttaaacatga gagcttagta cgttaaacat gagagcttag tacgtgaaac atgagagctt
     2041 agtacgtact atcaacaggt tgaactgcgg atcttgcggc cgcaaaaatt aaaaatgaag
     2101 ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat
     2161 cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc
     2221 cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat
     2281 accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag
     2341 ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg
     2401 ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc
     2461 tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca
     2521 acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg
     2581 tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc
     2641 actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta
     2701 ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc
     2761 aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg
     2821 ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc
     2881 cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc
     2941 aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat
     3001 actcatactc ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag
     3061 cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc
     3121 ccgaaaagtg ccacctgcat cgatggcccc ccgatggtag tgtggggtct ccccatgcga
     3181 gagtagggaa ctgccaggca tcaaataaaa cgaaaggctc agtcgaaaga ctgggccttt
     3241 cgttttatct gttgtttgtc ggtgaacgct ctcctgagta ggacaaatcc gccgggagcg
     3301 gatttgaacg ttgcgaagca acggcccgga gggtggcggg caggacgccc gccataaact
     3361 gccaggcatc aaattaagca gaaggccatc ctgacggatg gcctttttgc gtggccagtg
     3421 ccaagcttgc atgc
//

