LOCUS       sequence-83696-e         984 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  IPTG-inducible expression of Z-NGFP positive control for in vivo 
            split GFP assembly assay.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 984)
  AUTHORS   Magliery TJ, Wilson CG, Pan W, Mishler D, Ghosh I, Hamilton AD, 
            Regan L
  TITLE     Detecting protein-protein interactions with a green fluorescent 
            protein fragment reassembly trap: scope and mechanism.
  JOURNAL   J Am Chem Soc. 2005 Jan 12;127(1):146-57.
  PUBMED    15631464
REFERENCE   2  (bases 1 to 984)
  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: "J Am Chem 
            Soc."; date: "2005-01-12"; volume: "127(1)"; pages: "146-57"
COMMENT     Sequence Label: pET11a-Z-NGFP
FEATURES             Location/Qualifiers
     source          1..984
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     primer_bind     62..81
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        62..80
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    81..105
                     /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             120..142
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             159..176
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(210..238)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     CDS             726..737
                     /codon_start=1
                     /product="WELQut protease recognition and cleavage site"
                     /label=WELQut site
                     /translation="WELQ"
     primer_bind     complement(815..833)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      829..876
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
ORIGIN
        1 gccggtgatg ccggccacga tgcgtccggc gtagaggatc gagatctcga tcccgcgaaa
       61 ttaatacgac tcactatagg ggaattgtga gcggataaca attcccctct agaaataatt
      121 ttgtttaact ttaagaagga gatatacata tggctagcca tcaccaccat caccatggcg
      181 cgagcaaagg agaagaactc ttcactggag ttgtcccaat tcttgttgaa ttagatggtg
      241 atgttaacgg ccacaagttc tctgtcagtg gagagggtga aggtgatgca acatacggaa
      301 aacttaccct gaagttcatc tgcactactg gcaaactgcc tgttccatgg ccaacactag
      361 tcactactct gtgctatggt gttcaatgct tttcaagata cccggatcat atgaaacggc
      421 atgacttttt caagagtgcc atgcccgaag gttatgtaca ggaaaggacc atcttcttca
      481 aagatgacgg caactacaag acacgtgctg aagtcaagtt tgaaggtgat acccttgtta
      541 atagaatcga gttaaaaggt attgacttca aggaagatgg caacattctg ggacacaaat
      601 tggaatacaa ctataactca cacaacgttc tcatcatggc agacaaacaa ggtggctctg
      661 gctctggctc gagcgccctc aaaaaagaat tgcaggcaaa caaaaaagaa cttgcgcagc
      721 tgaagtggga gttacaagct ctgaaaaagg aactggcgca gtaaggatcc ggctgctaac
      781 aaagcccgaa aggaagctga gttggctgct gccaccgctg agcaataact agcataaccc
      841 cttggggcct ctaaacgggt cttgaggggt tttttgctga aaggaggaac tatatccgga
      901 tatcccgcaa gaggcccggc agtaccggca taaccaagcc tatgcctaca gcatccaggg
      961 tgacggtgcc gaggatgacg atga
//
