LOCUS       sequence-32154-e         788 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 788)
  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 788)
  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: pOPINM
FEATURES             Location/Qualifiers
     source          1..788
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     protein_bind    66..87
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(102..107,108..125,126..132)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    140..156
                     /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     145..167
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     164..180
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     164..180
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             176..349
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITDSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEART
                     DRPSQQLRSLNGE"
     primer_bind     complement(196..213)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(196..212)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(205..227)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     CDS             379..396
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(486..505)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    551..606
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(605..624)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     terminator      694..741
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
ORIGIN
        1 tctggaagtt ctgtttcagg gtaccgtcga cccgactgga aagcgggcag tgagcgcaac
       61 gcaattaatg tgagttagct cactcattag gcaccccagg ctttacactt tatgcttccg
      121 gctcgtatgt tgtgtggaat tgtgagcgga taacaatttc acacaggaaa cagctatgac
      181 catgattacg gattcactgg ccgtcgtttt acaacgtcgt gactgggaaa accctggcgt
      241 tacccaactt aatcgccttg cagcacatcc ccctttcgcc agctggcgta atagcgaaga
      301 ggcccgcacc gatcgccctt cccaacagtt gcgcagcctg aatggcgaat ggcgccaagc
      361 tttctagacc agtttaaaca tcaccatcac catcactaag tgattaacct caggtgcagg
      421 ctgcctatca gaaggtggtg gctggtgtgg ccaatgccct ggctcacaaa taccactgag
      481 atcgatcttt ttccctctgc caaaaattat ggggacatca tgaagcccct tgagcatctg
      541 acttctggct aataaaggaa atttattttc attgcaatag tgtgttggaa ttttttgtgt
      601 ctctcactcg gaaggacata tgggagggca aatcatttaa aacatcagaa tgagtatttg
      661 gtttagagtt tggcaacata tgcccatatg taactagcat aaccccttgg ggcctctaaa
      721 cgggtcttga ggggtttttt gctgaaagca tgcggaggaa attctccttg aagtttccct
      781 ggtgttca
//
