LOCUS       sequence-91266-e         535 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Mammalian expression vector for parallel SLIC cloning containing 
            N-terminal HA, 8x His and FLAG tags. C-terminal tags can be added 
            during SLIC..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 535)
  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 535)
  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: pCoofy50
FEATURES             Location/Qualifiers
     source          1..535
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     CDS             3..26
                     /codon_start=1
                     /product="8xHis affinity tag"
                     /label=8xHis
                     /translation="HHHHHHHH"
     CDS             36..59
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             66..89
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     RBS             126..134
                     /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             136..444
                     /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     323..342
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     CDS             448..477
                     /codon_start=1
                     /product="10xHis affinity tag"
                     /label=10xHis
                     /translation="HHHHHHHHHH"
ORIGIN
        1 cccatcacca tcaccatcac catcacggtg gttccgatta caaggatgac gatgacaagg
       61 aattcctgga agttctgttc caggggccct tgacaacata aaaactttgt gttatacttg
      121 taacgtaagg aggtaatgat tcagtttaag gtttacacct ataaaagaga gagccgttat
      181 cgtctgtttg tggatgtaca gagtgatatt attgacacgc ccgggcgacg gatggtgatc
      241 cccctggcca gtgcacgtct gctgtcagat aaagtctccc gtgaacttta cccggtggtg
      301 catatcgggg atgaaagctg gcgcatgatg accaccgata tggccagtgt gccggtctcc
      361 gttatcgggg aagaagtggc tgatctcagc caccgcgaaa atgacatcaa aaacgccatt
      421 aacctgatgt tctggggaat ataagagcat catcatcatc accaccacca ccaccactga
      481 gcggccgctt tcgaatctag aaattcaccc caccagtgca ggctgcctat cagaa
//
