LOCUS       sequence-32151-e         676 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 676)
  AUTHORS   Berrow NS, Alderton D, Sainsbury S, Nettleship J, Assenberg R, 
            Rahman N, Stuart DI, Owens RJ
  TITLE     A versatile ligation-independent cloning method suitable for 
            high-throughput expression screening applications.
  JOURNAL   Nucleic Acids Res. 2007  . 35(6):e45.
  PUBMED    17317681
REFERENCE   2  (bases 1 to 676)
  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: "Nucleic 
            Acids Res. 2007  . 35(6):e45."
COMMENT     Sequence Label: pOPINE
FEATURES             Location/Qualifiers
     source          1..676
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     CDS             90..107
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     protein_bind    160..181
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(196..201,202..219,220..226)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    234..250
                     /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)."
     primer_bind     239..261
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     258..274
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     258..274
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             270..443
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITDSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEART
                     DRPSQQLRSLNGE"
     primer_bind     complement(290..307)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(290..306)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(299..321)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     CDS             473..490
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(580..599)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
ORIGIN
        1 agaattatta tcaaatcatt tgtatattaa ttaaaatact atactgtaaa ttacatttta
       61 tttacaatca aaggagatat accatggcac accatcacca ccatcacagc agcggtaccg
      121 tcgacccgac tggaaagcgg gcagtgagcg caacgcaatt aatgtgagtt agctcactca
      181 ttaggcaccc caggctttac actttatgct tccggctcgt atgttgtgtg gaattgtgag
      241 cggataacaa tttcacacag gaaacagcta tgaccatgat tacggattca ctggccgtcg
      301 ttttacaacg tcgtgactgg gaaaaccctg gcgttaccca acttaatcgc cttgcagcac
      361 atcccccttt cgccagctgg cgtaatagcg aagaggcccg caccgatcgc ccttcccaac
      421 agttgcgcag cctgaatggc gaatggcgcc aagctttcta gaccagttta aacatcacca
      481 tcaccatcac taagtgatta acctcaggtg caggctgcct atcagaaggt ggtggctggt
      541 gtggccaatg ccctggctca caaataccac tgagatcgat ctttttccct ctgccaaaaa
      601 ttatggggac atcatgaagc cccttgagca tctgacttct ggctaataaa ggaaatttat
      661 tttcattgca atagtg
//
