LOCUS       sequence-62744-e         889 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 889)
  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 889)
  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: pCoofy2
FEATURES             Location/Qualifiers
     source          1..889
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     primer_bind     43..62
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        43..61
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    62..86
                     /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             101..123
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             131..457
                     /codon_start=1
                     /gene="trxA"
                     /product="E. coli thioredoxin"
                     /label=TrxA
                     /translation="MSDKIIHLTDDSFDTDVLKADGAILVDFWAEWCGPCKMIAPILDE
                     IADEYQGKLTVAKLNIDQNPGTAPKYGIRGIPTLLLFKNGEVAATKVGALSKGQLKEFL
                     DANLA"
     CDS             479..496
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             506..529
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     RBS             645..653
                     /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     842..861
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
ORIGIN
        1 atgcgtccgg cgtagaggat cgagatctcg atcccgcgaa attaatacga ctcactatag
       61 gggaattgtg agcggataac aattcccctc tagaaataat tttgtttaac tttaagaagg
      121 agatatacat atgagcgata aaattattca cctgactgac gacagttttg acacggatgt
      181 actcaaagcg gacggggcga tcctcgtcga tttctgggca gagtggtgcg gtccgtgcaa
      241 aatgatcgcc ccgattctgg atgaaatcgc tgacgaatat cagggcaaac tgaccgttgc
      301 aaaactgaac atcgatcaaa accctggcac tgcgccgaaa tatggcatcc gtggtatccc
      361 gactctgctg ctgttcaaaa acggtgaagt ggcggcaacc aaagtgggtg cactgtctaa
      421 aggtcagttg aaagagttcc tcgacgctaa cctggccggt tctggttctg gccatatgca
      481 ccatcatcat catcattctt ctggtctgga agttctgttc caggggccca tggactaatg
      541 aggtaccgga tccgaattcg agctccgtcg acaagcttga tatcgaattc ctgcagccca
      601 aactgcagtt gacaacataa aaactttgtg ttatacttgt aacgtaagga ggtaatgatt
      661 cagtttaagg tttacaccta taaaagagag agccgttatc gtctgtttgt ggatgtacag
      721 agtgatatta ttgacacgcc cgggcgacgg atggtgatcc ccctggccag tgcacgtctg
      781 ctgtcagata aagtctcccg tgaactttac ccggtggtgc atatcgggga tgaaagctgg
      841 cgcatgatga ccaccgatat ggccagtgtg ccggtctccg ttatcgggg
//
