LOCUS       sequence-91264-e         839 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Insect expression vector for parallel SLIC cloning containing 
            N-terminal 10x His and SumoStar tags. C-terminal tags can be added 
            during SLIC..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 839)
  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 839)
  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: pCoofy44
FEATURES             Location/Qualifiers
     source          1..839
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     CDS             64..93
                     /codon_start=1
                     /product="10xHis affinity tag"
                     /label=10xHis
                     /translation="HHHHHHHHHH"
     CDS             103..390
                     /codon_start=1
                     /gene="S. cerevisiae SMT3 (truncated and modified)"
                     /product="cleavable ubiquitin-like protein tag"
                     /label=SUMO
                     /note="modified to prevent cleavage by eukaryotic SUMO 
                     proteases"
                     /translation="DSEVNQEAKPEVKPEVKPETHINLKVSDGSSEIFFKIKKTTPLRR
                     LMEAFAKRQGKEMDSLTFLYDGIEIQADQTPEDLDMEDNDIIEAHREQIGG"
     CDS             397..420
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     RBS             457..465
                     /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             467..775
                     /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     654..673
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     CDS             779..808
                     /codon_start=1
                     /product="10xHis affinity tag"
                     /label=10xHis
                     /translation="HHHHHHHHHH"
ORIGIN
        1 aaacctataa atattccgga ttattcatac cgtcccacca tcgggcgcgg atccgaattc
       61 atgcaccatc accaccatca ccatcatcat cactccctgc aggactcaga agtcaatcaa
      121 gaagctaagc cagaggtcaa gccagaagtc aagcctgaga ctcacatcaa tttaaaggtg
      181 tccgatggat cttcagagat cttcttcaag atcaaaaaga ccactccttt aagaaggctg
      241 atggaagcgt tcgctaaaag acagggtaag gaaatggact ccttaacgtt cttgtacgac
      301 ggtattgaaa ttcaagctga tcagacccct gaagatttgg acatggagga taacgatatt
      361 attgaggctc atcgcgaaca gattggagga gaattcctgg aagttctgtt ccaggggccc
      421 ttgacaacat aaaaactttg tgttatactt gtaacgtaag gaggtaatga ttcagtttaa
      481 ggtttacacc tataaaagag agagccgtta tcgtctgttt gtggatgtac agagtgatat
      541 tattgacacg cccgggcgac ggatggtgat ccccctggcc agtgcacgtc tgctgtcaga
      601 taaagtctcc cgtgaacttt acccggtggt gcatatcggg gatgaaagct ggcgcatgat
      661 gaccaccgat atggccagtg tgccggtctc cgttatcggg gaagaagtgg ctgatctcag
      721 ccaccgcgaa aatgacatca aaaacgccat taacctgatg ttctggggaa tataagagca
      781 tcatcatcat caccaccacc accaccactg aagcgcttgg agccacccgc agttcgaaa
//
