LOCUS       sequence-65212-e        2291 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial targeting.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2291)
  AUTHORS   Alexeyev MF
  TITLE     The pKNOCK series of broad-host-range mobilizable suicide vectors 
            for gene knockout and targeted DNA insertion into the chromosome of 
            gram-negative bacteria.
  JOURNAL   Biotechniques. 1999 May;26(5):824-6, 828.
  PUBMED    10337469
REFERENCE   2  (bases 1 to 2291)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Biotechniques."; date: "1999-05"; volume: "26(5)"; pages: "824-6, 
            828"
COMMENT     Sequence Label: pKNOCK-Tc
FEATURES             Location/Qualifiers
     source          1..2291
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      33..421
                     /label=R6K gamma ori
                     /note="gamma replication origin from E. coli plasmid R6K; 
                     requires the R6K initiator protein pi for replication"
     primer_bind     454..470
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     454..470
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     primer_bind     complement(504..520)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(505..521)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     CDS             complement(654..1844)
                     /codon_start=1
                     /gene="tet"
                     /product="tetracycline efflux protein"
                     /label=TcR
                     /note="confers resistance to tetracycline"
                     /translation="MKSNNALIVILGTVTLDAVGIGLVMPVLPGLLRDIVHSDSIASHY
                     GVLLALYALMQFLCAPVLGALSDRFGRRPVLLASLLGATIDYAIMATTPVLWILYAGRI
                     VAGITGATGAVAGAYIADITDGEDRARHFGLMSACFGVGMVAGPVAGGLLGAISLHAPF
                     LAAAVLNGLNLLLGCFLMQESHKGERRPMPLRAFNPVSSFRWARGMTIVAALMTVLFIM
                     QLVGQVPAALWVIFGEDRFRWSATMIGLSLAVFGILHALAQAFVTGPATKRFGEKQAII
                     AGMAADALGYVLLAFATRGWMAFPIMILLASGGIGMPALQAMLSRQVDDDHQGQLQGSL
                     AALTSLTSIIGPLIVTAIYAASASTWNGLAWIVGAALYLVCLPALRRGAWSRATST"
     primer_bind     1473..1492
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     oriT            2044..2153
                     /note="incP origin of transfer"
ORIGIN
        1 attgatccgg ccacgatgcg tccggcgtag aggatctgaa gatcagcagt tcaacctgtt
       61 gatagtacgt actaagctct catgtttcac gtactaagct ctcatgttta acgtactaag
      121 ctctcatgtt taacgaacta aaccctcatg gctaacgtac taagctctca tggctaacgt
      181 actaagctct catgtttcac gtactaagct ctcatgtttg aacaataaaa ttaatataaa
      241 tcagcaactt aaatagcctc taaggtttta agttttataa gaaaaaaaag aatatataag
      301 gcttttaaag cttttaaggt ttaacggttg tggacaacaa gccagggatg taacgcactg
      361 agaagccctt agagcctctc aaagcaattt tgagtgacac aggaacactt aacggctgac
      421 atgggaattc ccctccaccg cggtggcggc cgctctagaa ctagtggatc ccccgggctg
      481 caggaattcg atatcaagct tatcgatacc gtcgacctcg agggggggcc cggtaccgag
      541 gacgcgtcag cttgatcaag ggttggtttg cgcattcaca gttctccgca agaattgatt
      601 ggctccaatt cttggagtgg tgaatccgtt agcgaggtgc cgccggcttc cattcaggtc
      661 gaggtggccc ggctccatgc accgcgacgc aacgcgggga ggcagacaag gtatagggcg
      721 gcgcctacaa tccatgccaa cccgttccat gtgctcgccg aggcggcata aatcgccgtg
      781 acgatcagcg gtccaatgat cgaagttagg ctggtaagag ccgcgagcga tccttgaagc
      841 tgtccctgat ggtcgtcatc tacctgcctg gacagcatgg cctgcaacgc gggcatcccg
      901 atgccgccgg aagcgagaag aatcataatg gggaaggcca tccagcctcg cgtcgcgaac
      961 gccagcaaga cgtagcccag cgcgtcggcc gccatgccgg cgataatggc ctgcttctcg
     1021 ccgaaacgtt tggtggcggg accagtgacg aaggcttgag cgagggcgtg caagattccg
     1081 aataccgcaa gcgacaggcc gatcatcgtc gcgctccagc gaaagcggtc ctcgccgaaa
     1141 atgacccaga gcgctgccgg cacctgtcct acgagttgca tgataaacaa gacagtcata
     1201 agtgcggcga cgatagtcat gccccgcgcc caccggaagg agctgactgg gttgaaggct
     1261 ctcaagggca tcggtcggcg ctctccctta tgcgactcct gcattaggaa gcagcccagt
     1321 agtaggttga ggccgttgag caccgccgcc gcaaggaatg gtgcatgtaa ggagatggcg
     1381 cccaacagtc ccccggccac ggggcctgcc accataccca cgccgaaaca agcgctcatg
     1441 agcccgaagt ggcgagcccg atcttcccca tcggtgatgt cggcgatata ggcgccagca
     1501 accgcacctg tggcgccggt gatgccggcc acgatgcgtc cggcgtagag aatccacagg
     1561 acgggtgtgg tcgccatgat cgcgtagtcg atagtggctc caagtagcga agcgagcagg
     1621 actgggcggc ggccaaagcg gtcggacagt gctccgagaa cgggtgcgca tagaaattgc
     1681 atcaacgcat atagcgctag cagcacgcca tagtgactgg cgatgctgtc ggaatggacg
     1741 atatcccgca agaggcccgg cagtaccggc ataaccaagc ctatgcctac agcatccagg
     1801 gtgacggtgc cgaggatgac gatgagcgca ttgttagatt tcatacacgg tgcctgactg
     1861 cgttagcaat ttaactgtga taaactaccg cattaaagct agcttatcga tgataagctg
     1921 tcaaacatga gaattgacgc gtcaattcga gggggatcaa ttccgtgata ggtgggctgc
     1981 ccttcctggt tggcttggtt tcatcagcca tccgcttgcc ctcatctgtt acgccggcgg
     2041 tagccggcca gcctcgcaga gcaggattcc cgttgagcac cgccaggtgc gaataaggga
     2101 cagtgaagaa ggaacacccg ctcgcgggtg ggcctacttc acctatcctg cccggctgac
     2161 gccgttggat acaccaagga aagtctacac gaaccctttg gcaaaatcct gtatatcgtg
     2221 cgaaaaagga tggatatacc gaaaaaatcg ctataatgac cccgaagcag ggttatgcag
     2281 cggaaaacgg a
//
