LOCUS       sequence-91262-e         709 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Insect expression vector for parallel SLIC cloning. C-terminal tags 
            can be added during SLIC..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 709)
  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 709)
  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: pCoofy41
FEATURES             Location/Qualifiers
     source          1..709
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     CDS             34..57
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     RBS             94..102
                     /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             104..412
                     /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     291..310
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     CDS             416..445
                     /codon_start=1
                     /product="10xHis affinity tag"
                     /label=10xHis
                     /translation="HHHHHHHHHH"
     CDS             join(449..454,455..478,479..514,515..538)
                     /codon_start=1
                     /product="two Strep-Tag(R)II moieties connected by a linker
                     for enhanced binding to Strep-Tactin(R), an engineered form
                     of streptavidin (Schmidt et al., 2013)"
                     /label=Twin-Strep-tag
                     /translation="SAWSHPQFEKGGGSGGGSGGSAWSHPQFEK"
     CDS             542..586
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     CDS             593..670
                     /codon_start=1
                     /product="calmodulin-binding peptide"
                     /label=CBP
                     /note="derived from skeletal muscle myosin light chain 
                     kinase; binds calmodulin with nanomolar affinity in the 
                     presence of calcium"
                     /translation="KRRWKKNFIAVSAANRFKKISSSGAL"
ORIGIN
        1 ggcgcggatc ccggtccgaa gcgcgcggaa ttcctggaag ttctgttcca ggggcccttg
       61 acaacataaa aactttgtgt tatacttgta acgtaaggag gtaatgattc agtttaaggt
      121 ttacacctat aaaagagaga gccgttatcg tctgtttgtg gatgtacaga gtgatattat
      181 tgacacgccc gggcgacgga tggtgatccc cctggccagt gcacgtctgc tgtcagataa
      241 agtctcccgt gaactttacc cggtggtgca tatcggggat gaaagctggc gcatgatgac
      301 caccgatatg gccagtgtgc cggtctccgt tatcggggaa gaagtggctg atctcagcca
      361 ccgcgaaaat gacatcaaaa acgccattaa cctgatgttc tggggaatat aagagcatca
      421 tcatcatcac caccaccacc accactgaag cgcttggagc cacccgcagt tcgaaaaagg
      481 tggaggttct ggcggtggat cgggaggttc agcgtggagc cacccgcagt tcgagaaata
      541 gaaagaaacc gctgctgcta aattcgaacg ccagcacatg gacagctaga tgaagcggcg
      601 gtggaagaaa aactttatcg ccgtgtccgc cgccaaccgg ttcaagaaga tcagcagctc
      661 tggcgccctg ctgagaggac caggtggacc cccggctgat cgacggcaa
//
