LOCUS       sequence-409922-        7207 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of the red fluorescent rogRFP2 redox sensor.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7207)
  AUTHORS   Radhakrishnan S, Norley J, Wendt S, LeRoy N, Hall H, Norcross S, 
            Doan S, Snaider J, MacVicar BA, Weake VM, Huang L, Tantama M
  TITLE     Neuron Activity Dependent Redox Compartmentation Revealed with a 
            Second Generation Red-Shifted Ratiometric Sensor.
  JOURNAL   ACS Chem Neurosci. 2020 Sep 2;11(17):2666-2678. doi: 
            10.1021/acschemneuro.0c00342. Epub 2020 Aug 21.
  PUBMED    32786310
REFERENCE   2  (bases 1 to 7207)
  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: 
            "10.1021/acschemneuro.0c00342"; journalName: "ACS Chem Neurosci"; 
            date: "2020-09-2- 2"; volume: "11"; issue: "17"; pages: "2666-2678"
COMMENT     Sequence Label: GW1-rogRFP2
FEATURES             Location/Qualifiers
     source          1..7207
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             join(189..191,192..194,195..707)
                     /codon_start=1
                     /product="N-terminal fragment of mVenus for use in 
                     bimolecular fluorescence complementation (BiFC) (Kodama and
                     Hu, 2010)"
                     /label=VN173
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFISTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNCHNVYIMADKQKNGIK
                     VNFKIRHNIE"
     primer_bind     complement(234..255)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(495..514)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     842..863
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     1277..1296
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     polyA_signal    1700..1834
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1737..1756)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1791..1810
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(1857..1873)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1857..1873)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1870..1892)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1881..1897
                     /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(1905..1911,1912..1929,1930..1935))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     primer_bind     complement(2087..2104)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2258..2846)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2338..2357)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3017..3808,3809..3877))
                     /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     3640..3659
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      complement(3934..4447)
                     /direction=LEFT
                     /label=M13 ori
                     /note="M13 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4129..4150)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4341..4360
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4619..4637
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4675..4697)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4797..4816
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5048..5070
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     5062..5079
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     5063..5079
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     enhancer        5645..6024
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        6025..6228
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     6179..6199
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     6225..6249
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     intron          6364..7190
                     /label=CMV intron A
                     /note="human cytomegalovirus intron A (Chapman et al., 
                     1991)"
ORIGIN
        1 aagcttcccg gccacagcaa tcgatccatg acacaaccag tgacaagcag tcccccagtg
       61 aaaccgcatc cgcatccgag tccgataccg ataaagattc tgaatcggag tgagtgccgc
      121 gtccgagagc gttccctgtc cacgtccacc atcggcggag caggtgtgcc tgaggcctct
      181 agaccaccat ggtgagcaag ggcgaggagc tgttcaccgg ggtggtgccc atcctggtcg
      241 agctggacgg cgacgtaaac ggccacaagt tcagcgtgtc cggcgagggc gagggcgatg
      301 ccacctacgg caagctgacc ctgaagttca tctccaccac cggcaagctg cccgtgccct
      361 ggcccaccct cgtgaccacc ctgacctacg gcgtgcagtg cttcagccgc taccccgacc
      421 acatgaagca gcacgacttc ttcaagtccg ccatgcccga aggctacgtc caggagcgca
      481 ccatcttctt caaggacgac ggcaactaca agacccgcgc cgaggtgaag ttcgagggcg
      541 acaccctggt gaaccgcatc gagctgaagg gcatcgactt caaggaggac ggcaacatcc
      601 tggggcacaa gctggagtac aactacaact gccacaacgt ctatatcatg gccgacaagc
      661 agaagaacgg catcaaggtg aacttcaaga tccgccacaa catcgaggac ggcagcgtgc
      721 agctcgccga ccactaccag cagaacaccc ccatcggcga cggccccgtg ctgctgcccg
      781 acaaccacta cctgagcacc tgctccgccc tgagcaaaga ccccaacgag aagcgcgatc
      841 acatggtcct gctggagttc gtgaccgctg caggtggaga ggaaaacaac atggccatca
      901 tcaaggagtt catgcgcttc aaggtgcaca tggagggctc cgtgaacggc cacgagttcg
      961 agatcgaggg cgagggcgag ggccgcccct acgaggcctt tcagaccgct aagctgaagg
     1021 tgaccaaggg tggccccctg cccttcgcct gggacatcct gtcccctcag ttcatgtacg
     1081 gctccaaggt ctacattaag cacccagccg acatccccga ctacttcaag ctgtccttcc
     1141 ccgagggctt caggtgggag cgcgtgatga acttcgagga cggcggcatt attcacgtta
     1201 accaggactc ctccctgcag gacggcgtgt tcatctacaa ggtgaagctg cgcggcacca
     1261 acttcccctc cgacggcccc gtaatgcaga agaagaccat gggctgggag gcctccgagg
     1321 agcggatgta ccccgaggac ggcgccctga agagcgagat caagaagagg ctgaagctga
     1381 aggacggcgg ccactacgcc gccgaggtca agaccaccta caaggccaag aagcccgtgc
     1441 agctgcccgg cgcctacatc gtcgacatca agttggacat cgtgtcccac aacgaggact
     1501 acaccatcgt ggaacagtac gaacgcgccg agggtcgtca ctctaccggc ggcatggacg
     1561 agctgtacaa gtaataaact cgacgaattc attgatcata atcagccata ccacatttgt
     1621 agaggtttta cttgctttaa aaaacctccc acacctcccc ctgaacctga aacataaaat
     1681 gaatgcaatt gttgttgtta acttgtttat tgcagcttat aatggttaca aataaagcaa
     1741 tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc
     1801 caaactcatc aatgtatctt atcatgtctg gatccgtacc gagctcgcgt aatcatgtca
     1861 tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat acgagccgga
     1921 agcataaagt gtaaagcctg gggtgcctaa tgagtgagct aactacatta attgcgttgc
     1981 gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc
     2041 aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact
     2101 cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac
     2161 ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa
     2221 aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg
     2281 acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa
     2341 gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc
     2401 ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac
     2461 gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac
     2521 cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg
     2581 taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt
     2641 atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa
     2701 cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct
     2761 cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga
     2821 ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg
     2881 ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct
     2941 tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt
     3001 aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc
     3061 tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg
     3121 gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag
     3181 atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt
     3241 tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag
     3301 ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt
     3361 ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca
     3421 tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg
     3481 ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat
     3541 ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta
     3601 tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca
     3661 gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct
     3721 taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat
     3781 cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa
     3841 agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt caataagcgg
     3901 ccgcggccat gccggccact agtctcgagg gccaccccgg ttgataatca gaaaagcccc
     3961 aaaaacagga agattgtata agcaaatatt taaattgtaa gcgttaatat tttgttaaaa
     4021 ttcgcgttaa atttttgtta aatcagctca ttttttaacc aataggccga aatcggcaaa
     4081 atcccttata aatcaaaaga atagaccgag atagggttga gtgttgttcc agtttggaac
     4141 aagagtccac tattaaagaa cgtggactcc aacgtcaaag ggcgaaaaac cgtctatcag
     4201 ggcgatggcc cactacgtga accatcaccc taatcaagtt ttttggggtc gaggtgccgt
     4261 aaagcactaa atcggaaccc taaagggagc ccccgattta gagcttgacg gggaaagccg
     4321 gcgaacgtgg cgagaaagga agggaagaaa gcgaaaggag cgggcgctag ggcgctggca
     4381 agtgtagcgg tcacgctgcg cgtaaccacc acacccgccg cgcttaatgc gccgctacag
     4441 ggcgcgtggg gatcactcga gttattattg aagcatttat cagggttatt gtctcatgag
     4501 cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc
     4561 ccgaaaagtg ccacctgacg tctaagaaac cattattatc atgacattaa cctataaaaa
     4621 taggcgtatc acgaggccct ttcgtctcgc gcgtttcggt gatgacggtg aaaacctctg
     4681 acacatgcag ctcccggaga cggtcacagc ttgtctgtaa gcggatgccg ggagcagaca
     4741 agcccgtcag ggcgcgtcag cgggtgttgg cgggtgtcgg ggctggctta actatgcggc
     4801 atcagagcag attgtactga gagtgcacca tatgaagacg tcgcctcctc actacttctg
     4861 gaatagctca gaggccgagg cggcctcggc ctctgcataa ataaaaaaaa ttagtcaggg
     4921 cgccattcgc cattcaggct gcgcaactgt tgggaagggc gatcggtgcg ggcctcttcg
     4981 ctattacgcc agctggcgaa agggggatgt gctgcaaggc gattaagttg ggtaacgcca
     5041 gggttttccc agtcacgacg ttgtaaaacg acggccagtg cccgtgtcaa ggacggtgac
     5101 tgcagtgaat aataaaatgt gtgtttgtcc gaaatacgcg ttttgagatt tctgtcgccg
     5161 actaaattca tgtcgcgcga tagtggtgtt tatcgccgat agagatggcg atattggaaa
     5221 aatcgatatt tgaaaatatg gcatattgaa aatgtcgccg atgtgagttt ctgtgtaact
     5281 gatatcgcca tttttccaaa agtgattttt gggcatacgc gatatctggc gatagcgctt
     5341 atatcgttta cgggggatgg cgatagacga ctttggtgac ttgggcgatt ctgtgtgtcg
     5401 caaatatcgc agtttcgata taggtgacag acgatatgag gctatatcgc cgatagaggc
     5461 gacatcaagc tggcacatgg ccaatgcata tcgatctata cattgaatca atattggcca
     5521 ttagccatat tattcattgg ttatatagca taaatcaata ttggctattg gccattgcat
     5581 acgttgtatc catatcataa tatgtacatt tatattggct catgtccaac attaccgcca
     5641 tgttgacatt gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat
     5701 agcccatata tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg
     5761 cccaacgacc cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata
     5821 gggactttcc attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta
     5881 catcaagtgt atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc
     5941 gcctggcatt atgcccagta catgacctta tgggactttc ctacttggca gtacatctac
     6001 gtattagtca tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga
     6061 tagcggtttg actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg
     6121 ttttggcacc aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg
     6181 caaatgggcg gtaggcgtgt acggtgggag gtctatataa gcagagctcg tttagtgaac
     6241 cgtcagatcg cctggagacg ccatccacgc tgttttgacc tccatagaag acaccgggac
     6301 cgatccagcc tccgcggccg ggaacggtgc attggaacgc ggattccccg tgccaagagt
     6361 gacgtaagta ccgcctatag agtctatagg cccaccccct tggcttctta tgcatgctat
     6421 actgtttttg gcttggggtc tatacacccc cgcttcctca tgttataggt gatggtatag
     6481 cttagcctat aggtgtgggt tattgaccat tattgaccac tcccctattg gtgacgatac
     6541 tttccattac taatccataa catggctctt tgccacaact ctctttattg gctatatgcc
     6601 aatacactgt ccttcagaga ctgacacgga ctctgtattt ttacaggatg gggtctcatt
     6661 tattatttac aaattcacat atacaacacc accgtcccca gtgcccgcag tttttattaa
     6721 acataacgtg ggatctccac gcgaatctcg ggtacgtgtt ccggacatgg gctcttctcc
     6781 ggtagcggcg gagcttctac atccgagccc tgctcccatg cctccagcga ctcatggtcg
     6841 ctcggcagct ccttgctcct aacagtggag gccagactta ggcacagcac gatgcccacc
     6901 accaccagtg tgccgcacaa ggccgtggcg gtagggtatg tgtctgaaaa tgagctcggg
     6961 gagcgggctt gcaccgctga cgcatttgga agacttaagg cagcggcaga agaagatgca
     7021 ggcagctgag ttgttgtgtt ctgataagag tcagaggtaa ctcccgttgc ggtgctgtta
     7081 acggtggagg gcagtgtagt ctgagcagta ctcgttgctg ccgcgcgcgc caccagacat
     7141 aatagctgac agactaacag actgttcctt tccatgggtc ttttctgcag tcaccgtcct
     7201 tgacacg
//
