LOCUS       sequence-109824-        4553 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial expression of His-tagged Grx1-roGFP2.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4553)
  AUTHORS   Gutscher M, Pauleau AL, Marty L, Brach T, Wabnitz GH, Samstag Y, 
            Meyer AJ, Dick TP
  TITLE     Real-time imaging of the intracellular glutathione redox potential.
  JOURNAL   Nat Methods. 2008 Jun;5(6):553-9. doi: 10.1038/nmeth.1212. Epub 2008
            May 11.
  PUBMED    18469822
REFERENCE   2  (bases 1 to 4553)
  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: 
            "10.1038/nmeth.1212"; journalName: "Nat Methods"; date: "2008-06"; 
            volume: "5"; issue: "6"; pages: "553-9"
COMMENT     Sequence Label: pQE-60 Grx1-roGFP2-His
FEATURES             Location/Qualifiers
     source          1..4553
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        join(10..24,25..30,31..47,48..54)
                     /label=T5 promoter
                     /note="bacteriophage T5 promoter for E. coli RNA 
                     polymerase, with embedded lac operator"
     protein_bind    30..46
                     /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    62..78
                     /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             97..108
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             join(535..537,538..1248)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FISTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNCHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTCSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(577..598)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(838..857)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1185..1206
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             1255..1272
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     terminator      1295..1389
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     primer_bind     complement(1499..1518)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     terminator      2161..2247
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     complement(2295..2317)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    2403..2543
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     2458..2477
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(2558..2575)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2729..3317)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2809..2828)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3488..4279,4280..4348))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     4111..4130
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4349..4453)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4521..4539
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 ctcgagaaat cataaaaaat ttatttgctt tgtgagcgga taacaattat aatagattca
       61 attgtgagcg gataacaatt tcacacagaa ttcattaaag aggagaaatt aaccatggct
      121 caagagtttg tgaactgcaa aatccagcct gggaaggtgg ttgtgttcat caagcccacc
      181 tgcccgtact gcaggagggc ccaagagatc ctcagtcaat tgcccatcaa acaagggctt
      241 ctggaatttg tcgatatcac agccaccaac cacactaacg agattcaaga ttatttgcaa
      301 cagctcacgg gagcaagaac ggtgcctcga gtctttattg gtaaagattg tataggcgga
      361 tgcagtgatc tagtctcttt gcaacagagt ggggaactgc tgacgcggct aaagcagatt
      421 ggagctctgc agactagtgg tggttcaggt ggtggtggtt caggtggtgg tggttcaggt
      481 ggaggaggat caggaggagg aggatcagga ggaggaggat caggaggaga attcgtgagc
      541 aagggcgagg agctgttcac cggggtggtg cccatcctgg tcgagctgga cggcgacgta
      601 aacggccaca agttcagcgt gtccggcgag ggcgagggcg atgccaccta cggcaagctg
      661 accctgaagt tcatctccac caccggcaag ctgcccgtgc cctggcccac cctcgtgacc
      721 accctgacct acggcgtgca gtgcttcagc cgctaccccg accacatgaa gcagcacgac
      781 ttcttcaagt ccgccatgcc cgaaggctac gtccaggagc gcaccatctt cttcaaggac
      841 gacggcaact acaagacccg cgccgaggtg aagttcgagg gcgacaccct ggtgaaccgc
      901 atcgagctga agggcatcga cttcaaggag gacggcaaca tcctggggca caagctggag
      961 tacaactaca actgccacaa cgtctatatc atggccgaca agcagaagaa cggcatcaag
     1021 gtgaacttca agatccgcca caacatcgag gacggcagcg tgcagctcgc cgaccactac
     1081 cagcagaaca cccccatcgg cgacggcccc gtgctgctgc ccgacaacca ctacctgagc
     1141 acctgctccg ccctgagcaa agaccccaac gagaagcgcg atcacatggt cctgctggag
     1201 ttcgtgaccg ccgccgggat cactctcggc atggacgagc tgtacaagag atctcatcac
     1261 catcaccatc actaagctta attagctgag cttggactcc tgttgataga tccagtaatg
     1321 acctcagaac tccatctgga tttgttcaga acgctcggtt gccgccgggc gttttttatt
     1381 ggtgagaatc caagctagct tggcgagatt ttcaggagct aaggaagcta aaatggagaa
     1441 aaaaatcact ggatatacca ccgttgatat atcccaatgg catcgtaaag aacattttga
     1501 ggcatttcag tcagttgctc aatgtaccta taaccagacc gttcagctgg atattacggc
     1561 ctttttaaag accgtaaaga aaaataagca caagttttat ccggccttta ttcacattct
     1621 tgcccgcctg atgaatgctc atccggaatt tcgtatggca atgaaagacg gtgagctggt
     1681 gatatgggat agtgttcacc cttgttacac cgttttccat gagcaaactg aaacgttttc
     1741 atcgctctgg agtgaatacc acgacgattt ccggcagttt ctacacatat attcgcaaga
     1801 tgtggcgtgt tacggtgaaa acctggccta tttccctaaa gggtttattg agaatatgtt
     1861 tttcgtctca gccaatccct gggtgagttt caccagtttt gatttaaacg tggccaatat
     1921 ggacaacttc ttcgcccccg ttttcaccat gcatgggcaa atattatacg caaggcgaca
     1981 aggtgctgat gccgctggcg attcaggttc atcatgccgt ctgtgatggc ttccatgtcg
     2041 gcagaatgct taatgaatta caacagtact gcgatgagtg gcagggcggg gcgtaatttt
     2101 tttaaggcag ttattggtgc ccttaaacgc ctggggtaat gactctctag cttgaggcat
     2161 caaataaaac gaaaggctca gtcgaaagac tgggcctttc gttttatctg ttgtttgtcg
     2221 gtgaacgctc tcctgagtag gacaaatccg ccgctctaga gctgcctcgc gcgtttcggt
     2281 gatgacggtg aaaacctctg acacatgcag ctcccggaga cggtcacagc ttgtctgtaa
     2341 gcggatgccg ggagcagaca agcccgtcag ggcgcgtcag cgggtgttgg cgggtgtcgg
     2401 ggcgcagcca tgacccagtc acgtagcgat agcggagtgt atactggctt aactatgcgg
     2461 catcagagca gattgtactg agagtgcacc atatgcggtg tgaaataccg cacagatgcg
     2521 taaggagaaa ataccgcatc aggcgctctt ccgcttcctc gctcactgac tcgctgcgct
     2581 cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca
     2641 cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga
     2701 accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc
     2761 acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg
     2821 cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat
     2881 acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt
     2941 atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc
     3001 agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg
     3061 acttatcgcc actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg
     3121 gtgctacaga gttcttgaag tggtggccta actacggcta cactagaagg acagtatttg
     3181 gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg
     3241 gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca
     3301 gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga
     3361 acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga
     3421 tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt
     3481 ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt
     3541 catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat
     3601 ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag
     3661 caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct
     3721 ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt
     3781 tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg
     3841 cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca
     3901 aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt
     3961 tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat
     4021 gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac
     4081 cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa
     4141 aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt
     4201 tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt
     4261 tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa
     4321 gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt
     4381 atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa
     4441 taggggttcc gcgcacattt ccccgaaaag tgccacctga cgtctaagaa accattatta
     4501 tcatgacatt aacctataaa aataggcgta tcacgaggcc ctttcgtctt cac
//
