LOCUS       sequence-117416-         834 bp    DNA     linear   UNA 14-APR-2025
DEFINITION  GFP^SEC^3xFlag vector with ccdB markers for cloning homology arms.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 834)
  AUTHORS   Dickinson DJ, Pani AM, Heppert JK, Higgins CD, Goldstein B
  TITLE     Streamlined Genome Engineering with a Self-Excising Drug Selection 
            Cassette.
  JOURNAL   Genetics. 2015 Jun 3. pii: genetics.115.178335.
  PUBMED    26044593
REFERENCE   2  (bases 1 to 834)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Genetics. 
            2015 Jun 3. pii: genetics.115.178335."
COMMENT     Sequence Label: pDD282
FEATURES             Location/Qualifiers
     source          1..834
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     CDS             233..538
                     /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="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
                     VPRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     primer_bind     417..436
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     CDS             complement(586..609)
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             complement(613..636)
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             complement(640..663)
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     protein_bind    682..715
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
ORIGIN
        1 gaggtatgct atgaagcagc gtattacagt gacagttgac agcgacagct atcagttgct
       61 caaggcatat atgatgtcaa tatctccggt tcggtaagca caaccatgca gaatgaagcc
      121 cgtcgtctgc gtgccgaacg ctggaaagcg gaaaatcagg aagggatggc tgaggtcgcc
      181 cggtttattg aaatgaacgg ctcttttgcc gacgagaaca ggggctggtg aaatgcagtt
      241 taaggtttac acctataaaa gagagagccg ttatcgtctg tttgtggatg tacagagtga
      301 tattattgac acgcccgggc gacggatggt gatccccctg gccagtgcac gtctgctgtc
      361 agataaagtc ccccgtgaac tttacccggt ggtgcatatc ggggatgaaa gctggcgcat
      421 gatgaccacc gatatggcca gtgtgccggt ctccgttatc ggggaagaag tggctgatct
      481 cagccaccgc gaaaatgaca tcaaaaacgc cattaacctg atgttctggg gaatataaat
      541 gtcaggctcc cttatacaca gccagtctgc aggtcgaact agtctcttgt catcgtcatc
      601 cttgtaatca cgcttgtcgt cgtcgtcctt gtagtcacgc ttatcgtcat cgtctttata
      661 atcagatccg gctccctgaa aataacttcg tataatgtat gctatacgaa gttattaaaa
      721 attgcggtca taaactgaaa cgtaacatat gataaggtat tttgtgtgcg gttttttcta
      781 tgatgtttca aaaaaattta tcagaagtaa aaaacataag aagtgtacac agaa
//
