LOCUS       sequence-26217-e         859 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  natural linear DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 859)
  AUTHORS   Wu S, Ying G, Wu Q, Capecchi MR
  TITLE     A protocol for constructing gene targeting vectors: generating 
            knockout mice for the cadherin family and beyond.
  JOURNAL   Nat Protoc. 2008  . 3(6):1056-76.
  PUBMED    18546598
REFERENCE   2  (bases 1 to 859)
  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: "Nat Protoc.
            2008  . 3(6):1056-76."
COMMENT     Sequence Label: pStart-K
FEATURES             Location/Qualifiers
     source          1..859
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     protein_bind    15..114
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             complement(143..448)
                     /codon_start=1
                     /gene="ccdB"
                     /product="CcdB, a bacterial toxin that poisons DNA gyrase"
                     /label=ccdB
                     /note="Plasmids containing the ccdB gene cannot be 
                     propagated in standard E. coli strains."
                     /translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRIVIPLASARLLSDK
                     VSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     primer_bind     complement(245..264)
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     protein_bind    complement(603..627)
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(722..745)
                     /label=pENTR-F
                     /note="pENTR vectors, forward primer"
ORIGIN
        1 ctgcagctgg nnggcaaata atgattttat tttgactgat agtgacctgt tcgttgcnac
       61 aaattgataa gcaatgcttt cttataatgc caactttgta caagaaagct gggtctagat
      121 atctcgagtg cggccgcgaa ttctatattc cccagaacat caggttaatg gcgtttttga
      181 tgtcattttc gcggtggctg agatcagcca cttcttcccc gataacggag accggcacac
      241 tggccatatc ggtggtcatc atgcgccagc tttcatcccc gatatgcacc accgggtaaa
      301 gttcacggga gactttatct gacagcagac gtgcactggc cagggggatc actatccgtc
      361 gcccgggcgt gtcaataata tcactctgta catccacaaa cagacgataa cggctctctc
      421 ttttataggt gtaaacctta aactgcattc tagaagcaca cctctttttg acatacttcg
      481 ggtatacata tcagtatata ttcttatacc gcaaaaatca gcgcgcaaat acgcatactg
      541 ttatctggct tttagtaagc gaattcggta ccggatccag tcgactgaat tggttccttt
      601 aaagcctgct tttttgtaca aagttggcat tataaaaaag cattgcttat caatttgttg
      661 caacgaacag gtcactatca nncnnannnn nnntcattat ttggggcccg agcttaagta
      721 actaactaac aggaagagtt tgtagaaacg caaaaaggcc atnngtcang atggccttct
      781 gcttagntng atgccngnca gttnatggcg ggcgtcctgc ccgncacctc ngggccgttg
      841 cttcacannn tnnantncg
//
