LOCUS       sequence-72281-e        1133 bp    DNA     linear   UNA 15-APR-2025
DEFINITION  Bact exp vector encoding for the 6XHis tagged insensitive variant of
            the fluorescent redox probe roGFP with mut Cys147 to Serine, del 
            insertion147b and mutated Cys204 to Glut.Referred to as refGFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 1133)
  AUTHORS   Avezov E, Cross BC, Kaminski Schierle GS, Winters M, Harding HP, 
            Melo EP, Kaminski CF, Ron D
  TITLE     Lifetime imaging of a fluorescent protein sensor reveals surprising 
            stability of ER thiol redox.
  JOURNAL   J Cell Biol. 2013 Apr 15;201(2):337-49. doi: 10.1083/jcb.201211155.
  PUBMED    23589496
REFERENCE   2  (bases 1 to 1133)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: "10.1083/jcb"; 
            journalName: "J Cell Biol"; date: "2013-04-15- 15"; volume: "201"; 
            issue: "2"; pages: "337-49"
COMMENT     Sequence Label: refGFP1_pQE30
FEATURES             Location/Qualifiers
     source          1..1133
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     primer_bind     27..45
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     protein_bind    89..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)."
     protein_bind    121..137
                     /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             156..167
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             186..203
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(240..268)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     terminator      1022..1116
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
ORIGIN
        1 ttattatcat gacattaacc tataaaaata ggcgtatcac gaggcccttt cgtcttcacc
       61 tcgagaaatc ataaaaaatt tatttgcttt gtgagcggat aacaatcaca atagattcaa
      121 ttgtgagcgg ataacaattt cacacagaat tcattaaaga ggagaaatta actatgagag
      181 gatcgcatca ccatcaccat cacggatcca tgagtaaagg agaagaactt ttcactggag
      241 ttgtcccaat tcttgttgaa ttagatggtg atgttaatgg gcacaaattt tctgtcagtg
      301 gagagggtga aggtgatgca acatacggaa aacttaccct taaatttatt tccactactg
      361 gaaaactacc tgttccatgg ccaacacttg tcactactct gagttatggt gttcaatgct
      421 tttcaagata cccagatcat atgaaacggc atgacttttt caagagtgcc atgcccgaag
      481 gttatgtaca ggaaagaact ataagcttca aggacgacgg gaattacaag acacgggccg
      541 aagtgaagtt tgagggagat actctggtga acaggattga actgaaaggg attgatttca
      601 aggaggacgg aaacatcctg ggccacaaac tggaatacaa ctacaacagt cataacgtgt
      661 acatcatggc cgataagcag aaaaacggga ttaaggctaa cttcaagacc cggcacaaca
      721 tcgaggacgg ctccgtgcag ctggcagatc attaccagca gaacacccca attggcgacg
      781 gacccgtcct gctgcctgat aatcactatc tgagcacaca gtccgccctg tctaaggacc
      841 ccaatgaaaa aagagatcat atggtcctgc tggaatttgt caccgccgca gggattactc
      901 acggaatgga tgaactgtat aaggacgaac tgtgataaag atctggtacc tcgagattgg
      961 gatcggatcc cgggcccgtc gactgcagag gcctgcatgc aagcttaatt agctgagctt
     1021 ggactcctgt tgatagatcc agtaatgacc tcagaactcc atctggattt gttcagaacg
     1081 ctcggttgcc gccgggcgtt ttttattggt gagaatccaa gctagcttgg cga
//
