LOCUS       sequence-324305-        5146 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Fluorescent biosensor for 2GSH/GSSG ratio.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5146)
  AUTHORS   Shokhina AG, Kostyuk AI, Ermakova YG, Panova AS, Staroverov DB, 
            Egorov ES, Baranov MS, van Belle GJ, Katschinski DM, Belousov VV, 
            Bilan DS
  TITLE     Red fluorescent redox-sensitive biosensor Grx1-roCherry.
  JOURNAL   Redox Biol. 2019 Feb;21:101071. doi: 10.1016/j.redox.2018.101071. 
            Epub 2018 Dec 7.
  PUBMED    30576927
REFERENCE   2  (bases 1 to 5146)
  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: "Redox 
            Biol."; date: "2019-02"; volume: "21:101071. doi"; pages: " 
            10.1016/j.redox.2018.101071. Epub 2018 Dec 7"
COMMENT     Sequence Label: pCS2+ Grx1-roCherry
FEATURES             Location/Qualifiers
     source          1..5146
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        35..53
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     35..52
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     CDS             458..1168
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="VSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEGT
                     QTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFE
                     DGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWECTESERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNICLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(600..618)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     868..887
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     polyA_signal    1198..1332
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1222..1241)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1276..1295
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(1445..1465)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(1445..1463)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(1484..1500)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1484..1500)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1497..1519)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1508..1524
                     /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)."
     promoter        complement(join(1532..1538,1539..1556,1557..1562))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1577..1598
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1715..1732)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1886..2474)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1966..1985)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2645..3436,3437..3505))
                     /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     3268..3287
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3506..3610)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(3636..4091)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3773..3794)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3985..4004
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4162..5146
                     /label=CMV IE94 promoter
                     /note="enhancer/promoter region of simian cytomegalovirus 
                     major immediate early transcription unit IE94"
ORIGIN
        1 cgccattctg cctggggacg tcggagcaag cttgatttag gtgacactat agaatacaag
       61 ctacttgttc tttttgcagg atcccatcga tatggctcaa gagtttgtga actgcaaaat
      121 ccagcctggg aaggtggttg tgttcatcaa gcccacctgc ccgtactgca ggagggccca
      181 agagatcctc agtcaattgc ccatcaaaca agggcttctg gaatttgtcg atatcacagc
      241 caccaaccac actaacgaga ttcaagatta tttgcaacag ctcacgggag caagaacggt
      301 gcctcgagtc tttattggta aagattgtat aggcggatgc agtgatctag tctctttgca
      361 acagagtggg gaactgctga cgcggctaaa gcagattgga gctctgcaga cttccggaac
      421 tgggggcaac gcctctgacg gtggtgggtc tggtggtgtg agcaagggcg aggaggataa
      481 catggccatc atcaaggagt tcatgcgctt caaggtgcac atggagggct ccgtgaacgg
      541 ccacgagttc gagatcgagg gcgagggcga gggccgcccc tacgagggca cccagaccgc
      601 caagctgaag gtgaccaagg gtggccccct gcccttcgcc tgggacatcc tgtcccctca
      661 gttcatgtac ggctccaagg cctacgtgaa gcaccccgcc gacatccccg actacttgaa
      721 gctgtccttc cccgagggct tcaagtggga gcgcgtgatg aacttcgagg acggcggcgt
      781 ggtgaccgtg acccaggact cctccctgca ggacggcgag ttcatctaca aggtgaagct
      841 gcgcggcacc aacttcccct ccgacggccc cgtaatgcag aagaagacca tgggctggga
      901 gtgcaccgaa tccgagcgga tgtaccccga ggacggcgcc ctgaagggcg agatcaagca
      961 gaggctgaag ctgaaggacg gcggccacta cgacgctgag gtcaagacca cctacaaggc
     1021 caagaagccc gtgcagctgc ccggcgccta caacgtcaac atctgcttgg acatcacctc
     1081 ccacaacgag gactacacca tcgtggaaca gtacgaacgc gccgagggcc gccactccac
     1141 cggcggcatg gacgagctgt acaagtaatc tagaactata gtgagtcgta ttacgtagat
     1201 ccagacatga taagatacat tgatgagttt ggacaaacca caactagaat gcagtgaaaa
     1261 aaatgcttta tttgtgaaat ttgtgatgct attgctttat ttgtaaccat tataagctgc
     1321 aataaacaag ttaacaacaa caattgcatt cattttatgt ttcaggttca gggggaggtg
     1381 tgggaggttt tttaattcgc ggccgcggcg ccaatgcatt gggcccggta cccagctttt
     1441 gttcccttta gtgagggtta attgcgcgct tggcgtaatc atggtcatag ctgtttcctg
     1501 tgtgaaattg ttatccgctc acaattccac acaacatacg agccgggagc ataaagtgta
     1561 aagcctgggg tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg
     1621 ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga
     1681 gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg
     1741 tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag
     1801 aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc
     1861 gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca
     1921 aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt
     1981 ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc
     2041 tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc
     2101 tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc
     2161 ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact
     2221 tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg
     2281 ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta
     2341 tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca
     2401 aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa
     2461 aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg
     2521 aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc
     2581 ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg
     2641 acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat
     2701 ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg
     2761 gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa
     2821 taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca
     2881 tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc
     2941 gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt
     3001 cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa
     3061 aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat
     3121 cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct
     3181 tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga
     3241 gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag
     3301 tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga
     3361 gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca
     3421 ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg
     3481 cgacacggaa atgttgaata ctcatactct tcctttttca atattattga agcatttatc
     3541 agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag
     3601 gggttccgcg cacatttccc cgaaaagtgc cacctaaatt gtaagcgtta atattttgtt
     3661 aaaattcgcg ttaaattttt gttaaatcag ctcatttttt aaccaatagg ccgaaatcgg
     3721 caaaatccct tataaatcaa aagaatagac cgagataggg ttgagtgttg ttccagtttg
     3781 gaacaagagt ccactattaa agaacgtgga ctccaacgtc aaagggcgaa aaaccgtcta
     3841 tcagggcgat ggcccactac gtgaaccatc accctaatca agttttttgg ggtcgaggtg
     3901 ccgtaaagca ctaaatcgga accctaaagg gagcccccga tttagagctt gacggggaaa
     3961 gccggcgaac gtggcgagaa aggaagggaa gaaagcgaaa ggagcgggcg ctagggcgct
     4021 ggcaagtgta gcggtcacgc tgcgcgtaac caccacaccc gccgcgctta atgcgccgct
     4081 acagggcgcg tcccattcgc cattcaggct gcgcaactgt tgggaagggc gatcggtgcg
     4141 ggcctcttcg ctattacgcc agtcgaccgc caattcaata tggcgtatat ggactcatgc
     4201 caattcaata tggtggatct ggacctgtgc caattcaata tggcgtatat ggactcgtgc
     4261 caattcaata tggtggatct ggaccccagc caattcaata tggcggactt ggcaccatgc
     4321 caattcaata tggcggacct ggcactgtgc caactgggga ggggtctact tggcacggtg
     4381 ccaagtttga ggaggggtct tggccctgtg ccaagtccgc catattgaat tggcatggtg
     4441 ccaataatgg cggccatatt ggctatatgc caggatcaat atataggcaa tatccaatat
     4501 ggccctatgc caatatggct attggccagg ttcaatacta tgtattggcc ctatgccata
     4561 tagtattcca tatatgggtt ttcctattga cgtagatagc ccctcccaat gggcggtccc
     4621 atataccata tatggggctt cctaataccg cccatagcca ctcccccatt gacgtcaatg
     4681 gtctctatat atggtctttc ctattgacgt catatgggcg gtcctattga cgtatatggc
     4741 gcctccccca ttgacgtcaa ttacggtaaa tggcccgcct ggctcaatgc ccattgacgt
     4801 caataggacc acccaccatt gacgtcaatg ggatggctca ttgcccattc atatccgttc
     4861 tcacgccccc tattgacgtc aatgacggta aatggcccac ttggcagtac atcaatatct
     4921 attaatagta acttggcaag tacattacta ttggaagtac gccagggtac attggcagta
     4981 ctcccattga cgtcaatggc ggtaaatggc ccgcgatggc tgccaagtac atccccattg
     5041 acgtcaatgg ggaggggcaa tgacgcaaat gggcgttcca ttgacgtaaa tgggcggtag
     5101 gcgtgcctaa tgggaggtct atataagcaa tgctcgttta gggaac
//
