LOCUS       sequence-32156-e         749 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 749)
  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 749)
  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: pOPINJ
FEATURES             Location/Qualifiers
     source          1..749
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     protein_bind    77..98
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(113..118,119..136,137..143)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    151..167
                     /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     156..178
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     175..191
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     175..191
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             187..360
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITDSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEART
                     DRPSQQLRSLNGE"
     primer_bind     complement(207..224)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(207..223)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(216..238)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     CDS             390..407
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(497..516)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    562..617
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(616..635)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
ORIGIN
        1 ctgagcagcg gtctggaagt tctgtttcag ggtaccgtcg acccgactgg aaagcgggca
       61 gtgagcgcaa cgcaattaat gtgagttagc tcactcatta ggcaccccag gctttacact
      121 ttatgcttcc ggctcgtatg ttgtgtggaa ttgtgagcgg ataacaattt cacacaggaa
      181 acagctatga ccatgattac ggattcactg gccgtcgttt tacaacgtcg tgactgggaa
      241 aaccctggcg ttacccaact taatcgcctt gcagcacatc cccctttcgc cagctggcgt
      301 aatagcgaag aggcccgcac cgatcgccct tcccaacagt tgcgcagcct gaatggcgaa
      361 tggcgccaag ctttctagac cagtttaaac atcaccatca ccatcactaa gtgattaacc
      421 tcaggtgcag gctgcctatc agaaggtggt ggctggtgtg gccaatgccc tggctcacaa
      481 ataccactga gatcgatctt tttccctctg ccaaaaatta tggggacatc atgaagcccc
      541 ttgagcatct gacttctggc taataaagga aatttatttt cattgcaata gtgtgttgga
      601 attttttgtg tctctcactc ggaaggacat atgggagggc aaatcattta aaacatcaga
      661 atgagtattt ggtttagagt ttggcaacat atgcccatat gtaactagca taaccccttg
      721 gggcctctaa acgggtcttg aggggtttt
//
