LOCUS       sequence-65205-e        1859 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 1859)
  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 1859)
  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-Cm
FEATURES             Location/Qualifiers
     source          1..1859
                     /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        654..756
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             757..1416
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(823..842)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     oriT            1612..1721
                     /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 gacgcgtccg aataaatacc tgtgacggaa gatcacttcg cagaataaat aaatcctggt
      601 gtccctgttg ataccgggaa gccctgggcc aacttttggc gaaaatgaga cgttgatcgg
      661 cacgtaagag gttccaactt tcaccataat gaaataagat cactaccggg cgtatttttt
      721 gagttatcga gattttcagg agctaaggaa gctaaaatgg agaaaaaaat cactggatat
      781 accaccgttg atatatccca atggcatcgt aaagaacatt ttgaggcatt tcagtcagtt
      841 gctcaatgta cctataacca gaccgttcag ctggatatta cggccttttt aaagaccgta
      901 aagaaaaata agcacaagtt ttatccggcc tttattcaca ttcttgcccg cctgatgaat
      961 gctcatccgg aatttcgtat ggcaatgaaa gacggtgagc tggtgatatg ggatagtgtt
     1021 cacccttgtt acaccgtttt ccatgagcaa actgaaacgt tttcatcgct ctggagtgaa
     1081 taccacgacg atttccggca gtttctacac atatattcgc aagatgtggc gtgttacggt
     1141 gaaaacctgg cctatttccc taaagggttt attgagaata tgtttttcgt ctcagccaat
     1201 ccctgggtga gtttcaccag ttttgattta aacgtggcca atatggacaa cttcttcgcc
     1261 cccgttttca ccatgggcaa atattatacg caaggcgaca aggtgctgat gccgctggcg
     1321 attcaggttc atcatgccgt ttgtgatggc ttccatgtcg gcagaatgct taatgaatta
     1381 caacagtact gcgatgagtg gcagggcggg gcgtaatttt tttaaggcag ttattggtgc
     1441 ccttaaacgc ctggttgcta cgcctgaata agtgataata agcggatgaa tggcagaaat
     1501 tcggacgcgt caattcgagg gggatcaatt ccgtgatagg tgggctgccc ttcctggttg
     1561 gcttggtttc atcagccatc cgcttgccct catctgttac gccggcggta gccggccagc
     1621 ctcgcagagc aggattcccg ttgagcaccg ccaggtgcga ataagggaca gtgaagaagg
     1681 aacacccgct cgcgggtggg cctacttcac ctatcctgcc cggctgacgc cgttggatac
     1741 accaaggaaa gtctacacga accctttggc aaaatcctgt atatcgtgcg aaaaaggatg
     1801 gatataccga aaaaatcgct ataatgaccc cgaagcaggg ttatgcagcg gaaaacgga
//
