LOCUS       sequence-65277-e        1698 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 1698)
  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 1698)
  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-Gm
FEATURES             Location/Qualifiers
     source          1..1698
                     /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"
     promoter        join(559..564,565..581,582..587)
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     CDS             776..1309
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     oriT            1451..1560
                     /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 gacgcgtcaa ttctcgaatt gacataagcc tgttcggttc gtaaactgta atgcaagtag
      601 cgtatgcgct cacgcaactg gtccagaacc ttgaccgaac gcagcggtgg taacggcgca
      661 gtggcggttt tcatggcttg ttatgactgt ttttttgtac agtctatgcc tcgggcatcc
      721 aagcagcaag cgcgttacgc cgtgggtcga tgtttgatgt tatggagcag caacgatgtt
      781 acgcagcagc aacgatgtta cgcagcaggg cagtcgccct aaaacaaagt taggtggctc
      841 aagtatgggc atcattcgca catgtaggct cggccctgac caagtcaaat ccatgcgggc
      901 tgctcttgat cttttcggtc gtgagttcgg agacgtagcc acctactccc aacatcagcc
      961 ggactccgat tacctcggga acttgctccg tagtaagaca ttcatcgcgc ttgctgcctt
     1021 cgaccaagaa gcggttgttg gcgctctcgc ggcttacgtt ctgcccaagt ttgagcagcc
     1081 gcgtagtgag atctatatct atgatctcgc agtctccggc gagcaccgga ggcagggcat
     1141 tgccaccgcg ctcatcaatc tcctcaagca tgaggccaac gcgcttggtg cttatgtgat
     1201 ctacgtgcaa gcagattacg gtgacgatcc cgcagtggct ctctatacaa agttgggcat
     1261 acgggaagaa gtgatgcact ttgatatcga cccaagtacc gccacctaac aattcgttca
     1321 agccgagatc ggcttcccgg ccgacgcgtc aattcgaggg ggatcaattc cgtgataggt
     1381 gggctgccct tcctggttgg cttggtttca tcagccatcc gcttgccctc atctgttacg
     1441 ccggcggtag ccggccagcc tcgcagagca ggattcccgt tgagcaccgc caggtgcgaa
     1501 taagggacag tgaagaagga acacccgctc gcgggtgggc ctacttcacc tatcctgccc
     1561 ggctgacgcc gttggataca ccaaggaaag tctacacgaa ccctttggca aaatcctgta
     1621 tatcgtgcga aaaaggatgg atataccgaa aaaatcgcta taatgacccc gaagcagggt
     1681 tatgcagcgg aaaacgga
//
