LOCUS       sequence-65281-e        2098 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 2098)
  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 2098)
  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-Km
FEATURES             Location/Qualifiers
     source          1..2098
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     oriT            93..202
                     /note="incP origin of transfer"
     rep_origin      373..761
                     /label=R6K gamma ori
                     /note="gamma replication origin from E. coli plasmid R6K; 
                     requires the R6K initiator protein pi for replication"
     primer_bind     794..810
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     794..810
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     primer_bind     complement(844..860)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(845..861)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     CDS             1258..2052
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(1312..1331)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     1922..1941
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
ORIGIN
        1 ggggatcaat tccgtgatag gtgggctgcc cttcctggtt ggcttggttt catcagccat
       61 ccgcttgccc tcatctgtta cgccggcggt agccggccag cctcgcagag caggattccc
      121 gttgagcacc gccaggtgcg aataagggac agtgaagaag gaacacccgc tcgcgggtgg
      181 gcctacttca cctatcctgc ccggctgacg ccgttggata caccaaggaa agtctacacg
      241 aaccctttgg caaaatcctg tatatcgtgc gaaaaaggat ggatataccg aaaaaatcgc
      301 tataatgacc ccgaagcagg gttatgcagc ggaaaacgga attgatccgg ccacgatgcg
      361 tccggcgtag aggatctgaa gatcagcagt tcaacctgtt gatagtacgt actaagctct
      421 catgtttcac gtactaagct ctcatgttta acgtactaag ctctcatgtt taacgaacta
      481 aaccctcatg gctaacgtac taagctctca tggctaacgt actaagctct catgtttcac
      541 gtactaagct ctcatgtttg aacaataaaa ttaatataaa tcagcaactt aaatagcctc
      601 taaggtttta agttttataa gaaaaaaaag aatatataag gcttttaaag cttttaaggt
      661 ttaacggttg tggacaacaa gccagggatg taacgcactg agaagccctt agagcctctc
      721 aaagcaattt tgagtgacac aggaacactt aacggctgac atgggaattc ccctccaccg
      781 cggtggcggc cgctctagaa ctagtggatc ccccgggctg caggaattcg atatcaagct
      841 tatcgatacc gtcgacctcg agggggggcc cggtaccgag gacgcgtcga attaattccg
      901 ctagcttcac gctgccgcaa gcactcaggg cgcaagggct gctaaaggaa gcggaacacg
      961 tagaaagcca gtccgcagaa acggtgctga ccccggatga atgtcagcta ctgggctatc
     1021 tggacaaggg aaaacgcaag cgcaaagaga aagcaggtag cttgcagtgg gcttacatgg
     1081 cgatagctag actgggcggt tttatggaca gcaagcgaac cggaattgcc agctggggcg
     1141 ccctctggta aggttgggaa gccctgcaaa gtaaactgga tggctttctt gccgccaagg
     1201 atctgatggc gcaggggatc aagatctgat caagagacag gatgaggatc gtttcgcatg
     1261 attgaacaag atggattgca cgcaggttct ccggccgctt gggtggagag gctattcggc
     1321 tatgactggg cacaacagac aatcggctgc tctgatgccg ccgtgttccg gctgtcagcg
     1381 caggggcgcc cggttctttt tgtcaagacc gacctgtccg gtgccctgaa tgaactccaa
     1441 gacgaggcag cgcggctatc gtggctggcc acgacgggcg ttccttgcgc agctgtgctc
     1501 gacgttgtca ctgaagcggg aagggactgg ctgctattgg gcgaagtgcc ggggcaggat
     1561 ctcctgtcat ctcaccttgc tcctgccgag aaagtatcca tcatggctga tgcaatgcgg
     1621 cggctgcata cgcttgatcc ggctacctgc ccattcgacc accaagcgaa acatcgcatc
     1681 gagcgagcac gtactcggat ggaagccggt cttgtcgatc aggatgatct ggacgaagag
     1741 catcaggggc tcgcgccagc cgaactgttc gccaggctca aggcgcggat gcccgacggc
     1801 gaggatctcg tcgtgaccca tggcgatgcc tgcttgccga atatcatggt ggaaaatggc
     1861 cgcttttctg gattcatcga ctgtggccgg ctgggtgtgg cggaccgcta tcaggacata
     1921 gcgttggcta cccgtgatat tgctgaagag cttggcggcg aatgggctga ccgcttcctc
     1981 gtgctttacg gtatcgccgc tcccgattcg cagcgcatcg ccttctatcg ccttcttgac
     2041 gagttcttct gagcgggact ctggggttcg cggaattaat tcgacgcgtc aattcgag
//
