LOCUS       sequence-63541-e         880 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 880)
  AUTHORS   Scholz J, Besir H, Strasser C, Suppmann S
  TITLE     A new method to customize protein expression vectors for fast, 
            efficient and background free parallel cloning.
  JOURNAL   BMC Biotechnol. 2013 Feb 14;13(1):12.
  PUBMED    23410102
REFERENCE   2  (bases 1 to 880)
  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: "BMC 
            Biotechnol."; date: "2013-02-14"; volume: "13(1)"; pages: "12"
COMMENT     Sequence Label: pCoofy3
FEATURES             Location/Qualifiers
     source          1..880
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     CDS             complement(21..38)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             complement(58..366)
                     /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="MIQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSD
                     KVSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     primer_bind     complement(160..179)
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     RBS             368..376
                     /label=Shine-Dalgarno sequence
                     /note="full consensus sequence for ribosome-binding sites 
                     upstream of start codons in E. coli; complementary to a 
                     region in the 3' end of the 16S rRNA (Chen et al., 1994)"
     CDS             complement(492..515)
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     primer_bind     complement(542..564)
                     /label=pGEX 5'
                     /note="pGEX vectors, Glutathione-S-transferase, forward 
                     primer"
ORIGIN
        1 ttgttagcag ccggatctca gtggtggtgg tggtggtgct cgagtgcggc cgcaagctta
       61 tattccccag aacatcaggt taatggcgtt tttgatgtca ttttcgcggt ggctgagatc
      121 agccacttct tccccgataa cggagaccgg cacactggcc atatcggtgg tcatcatgcg
      181 ccagctttca tccccgatat gcaccaccgg gtaaagttca cgggagactt tatctgacag
      241 cagacgtgca ctggccaggg ggatcaccat ccgtcgcccg ggcgtgtcaa taatatcact
      301 ctgtacatcc acaaacagac gataacggct ctctctttta taggtgtaaa ccttaaactg
      361 aatcattacc tccttacgtt acaagtataa cacaaagttt ttatgttgtc aactgcagtt
      421 tgggctgcag gaattcgata tcaagcttgt cgacggagct cgaattcgga tccggtacct
      481 cattagtcca tgggcccctg gaacagaact tccagatccg attttggagg atggtcgcca
      541 ccaccaaacg tggcttgcca gccctgcaaa ggccatgcta tatacttgct ggatttcaag
      601 tacttatcaa tttgtgggat agcttcaata cgttttttaa aacaaactaa ttttgggaac
      661 gcatccaggc acattgggtc catgtataaa acaacatcaa gagcgtcata caacatgaag
      721 tcaggatggg ttacatgatc accatttaaa tatgttttat gacataaacg atcttcgaac
      781 attttcagca tttcaggtag cttgctaaga aaatcaactt tgagagtttc aaagtcttta
      841 ctatatgcaa ttctcgaaac accgtatcta atatccaaaa
//
