LOCUS       sequence-62746-e         928 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 928)
  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 928)
  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: pCoofy38
FEATURES             Location/Qualifiers
     source          1..928
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     primer_bind     53..72
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        53..71
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    72..96
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     RBS             111..133
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             141..467
                     /codon_start=1
                     /gene="trxA"
                     /product="E. coli thioredoxin"
                     /label=TrxA
                     /translation="MSDKIIHLTDDSFDTDVLKADGAILVDFWAEWCGPCKMIAPILDE
                     IADEYQGKLTVAKLNIDQNPGTAPKYGIRGIPTLLLFKNGEVAATKVGALSKGQLKEFL
                     DANLA"
     CDS             489..518
                     /codon_start=1
                     /product="10xHis affinity tag"
                     /label=10xHis
                     /translation="HHHHHHHHHH"
     CDS             528..551
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     RBS             667..675
                     /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)"
     primer_bind     864..883
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
ORIGIN
        1 gccggccacg atgcgtccgg cgtagaggat cgagatctcg atcccgcgaa attaatacga
       61 ctcactatag gggaattgtg agcggataac aattcccctc tagaaataat tttgtttaac
      121 tttaagaagg agatatacat atgagcgata aaattattca cctgactgac gacagttttg
      181 acacggatgt actcaaagcg gacggggcga tcctcgtcga tttctgggca gagtggtgcg
      241 gtccgtgcaa aatgatcgcc ccgattctgg atgaaatcgc tgacgaatat cagggcaaac
      301 tgaccgttgc aaaactgaac atcgatcaaa accctggcac tgcgccgaaa tatggcatcc
      361 gtggtatccc gactctgctg ctgttcaaaa acggtgaagt ggcggcaacc aaagtgggtg
      421 cactgtctaa aggtcagttg aaagagttcc tcgacgctaa cctggccggt tctggttctg
      481 gccatatgca ccatcatcac caccaccacc atcatcattc ttctggtctg gaagttctgt
      541 tccaggggcc catggactaa tgaggtaccg gatccgaatt cgagctccgt cgacaagctt
      601 gatatcgaat tcctgcagcc caaactgcag ttgacaacat aaaaactttg tgttatactt
      661 gtaacgtaag gaggtaatga ttcagtttaa ggtttacacc tataaaagag agagccgtta
      721 tcgtctgttt gtggatgtac agagtgatat tattgacacg cccgggcgac ggatggtgat
      781 ccccctggcc agtgcacgtc tgctgtcaga taaagtctcc cgtgaacttt acccggtggt
      841 gcatatcggg gatgaaagct ggcgcatgat gaccaccgat atggccagtg tgccggtctc
      901 cgttatcggg gaagaagtgg ctgatctc
//
