LOCUS       sequence-72301-e         957 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Bacterial expression vector encoding for the 6XHis tagged 
            insensitive variant of fluorescent redox probe roGFPiE with Cysteine
            204 to Serine mutation (roGFP C147S, E147b, C204).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 957)
  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 957)
  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"; doi: "10.1083/jcb"; 
            journalName: "J Cell Biol"; date: "2013-04-15- 15"; volume: "201"; 
            issue: "2"; pages: "337-49"
COMMENT     Sequence Label: roGFP_iE_C204S_pQE30
FEATURES             Location/Qualifiers
     source          1..957
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     protein_bind    11..27
                     /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    43..59
                     /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             78..89
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             108..125
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             132..848
                     /codon_start=1
                     /product="S65T variant of Aequorea victoria green 
                     fluorescent protein (Heim et al., 1995)"
                     /label=GFP (S65T)
                     /note="excitable with blue light"
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FISTTGKLPVPWPTLVTTLSYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTISFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNCESNVYIMADKQKNGIK
                     ANFKTRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTSSALSKDPNEKRDHMVLL
                     EFVTAAGITHGMDELYK"
     primer_bind     complement(162..190)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     788..808
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
ORIGIN
        1 tttatttgct ttgtgagcgg ataacaatca caatagattc aattgtgagc ggataacaat
       61 ttcacacaga attcattaaa gaggagaaat taactatgag aggatcgcat caccatcacc
      121 atcacggatc catgagtaaa ggagaagaac ttttcactgg agttgtccca attcttgttg
      181 aattagatgg tgatgttaat gggcacaaat tttctgtcag tggagagggt gaaggtgatg
      241 caacatacgg aaaacttacc cttaaattta tttccactac tggaaaacta cctgttccat
      301 ggccaacact tgtcactact ctgagttatg gtgttcaatg cttttcaaga tacccagatc
      361 atatgaaacg gcatgacttt ttcaagagtg ccatgcccga aggttatgta caggaaagaa
      421 ctataagctt caaagatgac gggaactaca agacacgtgc tgaagtcaag tttgaaggtg
      481 atacccttgt taatagaatc gagttaaaag gtattgattt taaagaagat ggaaacattc
      541 ttggacacaa attggaatac aactataact gcgagtccaa tgtatacatc atggcagaca
      601 aacaaaagaa tggaatcaaa gcgaacttca aaaccagaca caacattgaa gatggaagcg
      661 ttcaactagc agaccattat caacaaaata ctccaattgg cgatggccct gtccttttac
      721 cagacaacca ttacctgtcc acatcctctg ccctttcgaa agatcccaac gaaaagagag
      781 accacatggt ccttcttgag tttgtaacag ctgctgggat tacacatggc atggatgaac
      841 tatacaaata ataacgatcc gagctcgaga tctgcagcca agcttaatta gctgagcttg
      901 gactcctgtt gatagatcca gtaatgacct cagaactcca tctggatttg ttcagaa
//
