LOCUS       sequence-211579-        6883 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses thiol redox-sensitive ratiometric sensor roGFP in cytosol 
            of mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6883)
  AUTHORS   Waypa GB, Marks JD, Guzy R, Mungai PT, Schriewer J, Dokic D, 
            Schumacker PT
  TITLE     Hypoxia triggers subcellular compartmental redox signaling in 
            vascular smooth muscle cells.
  JOURNAL   Circ Res. 2010 Feb 19;106(3):526-35. doi: 
            10.1161/CIRCRESAHA.109.206334. Epub 2009 Dec 17.
  PUBMED    20019331
REFERENCE   2  (bases 1 to 6883)
  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: 
            "10.1161/CIRCRESAHA.109.206334"; journalName: "Circ Res"; date: 
            "2010-02-19- 19"; volume: "106"; issue: "3"; pages: "526-35"
COMMENT     Sequence Label: Cyto-roGFP
FEATURES             Location/Qualifiers
     source          1..6883
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(108..127)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     455..476
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    542..676
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(566..585)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     620..639
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(819..841)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     rep_origin      complement(3916..4504)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3996..4015)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4675..5466,5467..5535))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     5298..5317
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5536..5640)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5708..5726
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     repeat_region   5754..5856
                     /label=ITR
                     /note="inverted terminal repeat of human adenovirus 
                     serotype 5"
     misc_signal     5944..6094
                     /label=Ad5 Psi
                     /note="packaging signal for adenovirus serotype 5"
     enhancer        6123..6426
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        6427..6630
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     6581..6601
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      6679..6688
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(6685..6687,6688..6690,6691..6883,1..521)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFISTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNCHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTCSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(6730..6751)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
ORIGIN
        1 acggcgtgca gtgcttcagc cgctaccccg accacatgaa gcggcacgac ttcttcaagt
       61 ccgccatgcc cgaaggctac gtccaggagc gcaccatctt cttcaaggac gacggcaact
      121 acaagacccg cgccgaggtg aagttcgagg gcgacaccct ggtgaaccgc atcgagctga
      181 agggcatcga cttcaaggag gacggcaaca tcctggggca caagctggag tacaactaca
      241 actgccacaa cgtctatatc atggccgaca agcagaagaa cggcatcaag gtgaacttca
      301 agatccgcca caacatcgag gacggcagcg tgcagctcgc cgaccactac cagcagaaca
      361 cccccatcgg cgacggcccc gtgctgctgc ccgacaacca ctacctgagc acctgctccg
      421 ccctgagcaa agaccccaac gagaagcgcg atcacatggt cctgctggag ttcgtgaccg
      481 ccgccgggat cactctcggc atggacgagc tgtacaagta aagcggccgc cacagcgggg
      541 agatccagac atgataagat acattgatga gtttggacaa accacaacta gaatgcagtg
      601 aaaaaaatgc tttatttgtg aaatttgtga tgctattgct ttatttgtaa ccattataag
      661 ctgcaataaa caagttaaca acaacaattg cattcatttt atgtttcagg ttcaggggga
      721 ggtgtgggag gttttttaaa gcaagtaaaa cctctacaaa tgtggtatgg ctgattatga
      781 tccggctgcc tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg
      841 gagacggtca cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg
      901 tcagcgggtg ttggcgggtg tcggggcgca gccatgaggt cgactctagt ccccgcggtg
      961 gcagatctgg aaggtgctga ggtacgatga gacccgcacc aggtgcagac cctgcgagtg
     1021 tggcggtaaa catattagga accagcctgt gatgctggat gtgaccgagg agctgaggcc
     1081 cgatcacttg gtgctggcct gcacccgcgc tgagtttggc tctagcgatg aagatacaga
     1141 ttgaggtact gaaatgtgtg ggcgtggctt aagggtggga aagaatatat aaggtggggg
     1201 tcttatgtag ttttgtatct gttttgcagc agccgccgcc gccatgagca ccaactcgtt
     1261 tgatggaagc attgtgagct catatttgac aacgcgcatg cccccatggg ccggggtgcg
     1321 tcagaatgtg atgggctcca gcattgatgg tcgccccgtc ctgcccgcaa actctactac
     1381 cttgacctac gagaccgtgt ctggaacgcc gttggagact gcagcctccg ccgccgcttc
     1441 agccgctgca gccaccgccc gcgggattgt gactgacttt gctttcctga gcccgcttgc
     1501 aagcagtgca gcttcccgtt catccgcccg cgatgacaag ttgacggctc ttttggcaca
     1561 attggattct ttgacccggg aacttaatgt cgtttctcag cagctgttgg atctgcgcca
     1621 gcaggtttct gccctgaagg cttcctcccc tcccaatgcg gtttaaaaca taaataaaaa
     1681 accagactct gtttggattt ggatcaagca agtgtcttgc tgtctttatt taggggtttt
     1741 gcgcgcgcgg taggcccggg accagcggtc tcggtcgttg agggtcctgt gtattttttc
     1801 caggacgtgg taaaggtgac tctggatgtt cagatacatg ggcataagcc cgtctctggg
     1861 gtggaggtag caccactgca gagcttcatg ctgcggggtg gtgttgtaga tgatccagtc
     1921 gtagcaggag cgctgggcgt ggtgcctaaa aatgtctttc agtagcaagc tgattgccag
     1981 gggcaggccc ttggtgtaag tgtttacaaa gcggttaagc tgggatgggt gcatacgtgg
     2041 ggatatgaga tgcatcttgg actgtatttt taggttggct atgttcccag ccatatccct
     2101 ccggggattc atgttgtgca gaaccaccag cacagtgtat ccggtgcact tgggaaattt
     2161 gtcatgtagc ttagaaggaa atgcgtggaa gaacttggag acgcccttgt gacctccaag
     2221 attttccatg cattcgtcca taatgatggc aatgggccca cgggcggcgg cctgggcgaa
     2281 gatatttctg ggatcactaa cgtcatagtt gtgttccagg atgagatcgt cataggccat
     2341 ttttacaaag cgcgggcgga gggtgccaga ctgcggtata atggttccat ccggcccagg
     2401 ggcgtagtta ccctcacaga tttgcatttc ccacgctttg agttcagatg gggggatcat
     2461 gtctacctgc ggggcgatga agaaaacggt ttccggggta ggggagatca gctgggaaga
     2521 aagcaggttc ctgagcagct gcgacttacc gcagccggtg ggcccgtaaa tcacacctat
     2581 taccggctgc aactggtagt taagagagct gcagctgccg tcatccctga gcaggggggc
     2641 cacttcgtta agcatgtccc tgactcgcat gttttccctg accaaatccg ccagaaggcg
     2701 ctcgccgccc agcgatagca gttcttgcaa ggaagcaaag tttttcaacg gtttgagacc
     2761 gtccgccgta ggcatgcttt tgagcgtttg accaagcagt tccaggcggt cccacagctc
     2821 ggtcacctgc tctacggcat ctcgatccag catatctcct cgtttcgcgg gttggggcgg
     2881 ctttcgctgt acggcagtag tcggtgctcg tccagacggg ccagggtcat gtctttccac
     2941 gggcgcaggg tcctcgtcag cgtagtctgg gtcacggtga aggggtgcgc tccgggctgc
     3001 gcgctggcca gggtgcgctt gaggctggtc ctgctggtgc tgaagcgctg ccggtcttcg
     3061 ccctgcgcgt cggccaggta gcatttgacc atggtgtcat agtccagccc ctccgcggcg
     3121 tggcccttgg cgcgcagctt gcccttggag gaggcgccgc acgaggggca gtgcagactt
     3181 ttgagggcgt agagcttggg cgcgagaaat accgattccg gggagtaggc atccgcgccg
     3241 caggccccgc agacggtctc gcattccacg agccaggtga gctctggccg ttcggggtca
     3301 aaaaccaggt ttcccccatg ctttttgatg cgtttcttac ctctggtttc catgagccgg
     3361 tgtccacgct cggtgacgaa aaggctgtcc gtgtccccgt atacagactt gagaggcctg
     3421 tcctcgaccg atgcccttga gagccttcaa cccagtcagc tccttccggt gggcgcgggg
     3481 catgactatc gtcgccgcac ttatgactgt cttctttatc atgcaactcg taggacaggt
     3541 gccggcagcg ctctgggtca ttttcggcga ggaccgcttt cgctggagcg cgacgatgat
     3601 cggcctgtcg cttgcggtat tcggaatctt gcacgccctc gctcaagcct tcgtcactgg
     3661 tcccgccacc aaacgtttcg gcgagaagca ggccattatc gccggcatgg cggccgacgc
     3721 gctgggctac gtcttgctgg cgttcgcgac gcgaggctgg atggccttcc ccattatgat
     3781 tcttctcgct tccggcggca tcgggatgcc cgcgttgcag gccatgctgt ccaggcaggt
     3841 agatgacgac catcagggac agcttcaagg ccagcaaaag gccaggaacc gtaaaaaggc
     3901 cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg
     3961 ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg
     4021 aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt
     4081 tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt
     4141 gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg
     4201 cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact
     4261 ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt
     4321 cttgaagtgg tggcctaact acggctacac tagaaggaca gtatttggta tctgcgctct
     4381 gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac
     4441 cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc
     4501 tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg
     4561 ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta
     4621 aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca
     4681 atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc
     4741 ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc
     4801 tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc
     4861 agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat
     4921 taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt
     4981 tgccattgct gcaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc
     5041 cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag
     5101 ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt
     5161 tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac
     5221 tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg
     5281 cccggcgtca acacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat
     5341 tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc
     5401 gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc
     5461 tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa
     5521 atgttgaata ctcatactct tcctttttca atattattga agcatttatc agggttattg
     5581 tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg
     5641 cacttttcgg ggaaatgtgc cacctgacgt ctaagaaacc attattatca tgacattaac
     5701 ctataaaaat aggcgtatca cgaggccctt tcgtcttcaa ttaattaagc tagcatcatc
     5761 aataatatac cttattttgg attgaagcca atatgataat gagggggtgg agtttgtgac
     5821 gtggcgcggg gcgtgggaac ggggcgggtg acgtagtagt gtggcggaag tgtgatgttg
     5881 caagtgtggc ggaacacatg taagcgacgg atgtggcaaa agtgacgttt ttggtgtgcg
     5941 ccggtgtaca caggaagtga caattttcgc gcggttttag gcggatgttg tagtaaattt
     6001 gggcgtaacc gagtaagatt tggccatttt cgcgggaaaa ctgaataaga ggaagtgaaa
     6061 tctgaataat tttgtgttac tcatagcgcg taatatttgt ctagggagat cagcctgcag
     6121 gtcgttacat aacttacggt aaatggcccg cctggctgac cgcccaacga cccccgccca
     6181 ttgacgtcaa taatgacgta tgttcccata gtaacgccaa tagggacttt ccattgacgt
     6241 caatgggtgg agtatttacg gtaaactgcc cacttggcag tacatcaagt gtatcatatg
     6301 ccaagtacgc cccctattga cgtcaatgac ggtaaatggc ccgcctggca ttatgcccag
     6361 tacatgacct tatgggactt tcctacttgg cagtacatct acgtattagt catcgctatt
     6421 accatggtga tgcggttttg gcagtacatc aatgggcgtg gatagcggtt tgactcacgg
     6481 ggatttccaa gtctccaccc cattgacgtc aatgggagtt tgttttggca ccaaaatcaa
     6541 cgggactttc caaaatgtcg taacaactcc gccccattga cgcaaatggg cggtaggcgt
     6601 gtacggtggg aggtctatat aagcagagct cgtttagtga accgtcagat ggtaccgcgg
     6661 gcccgggatc caccggtcgc caccatggtg agcaagggcg aggagctgtt caccggggtg
     6721 gtgcccatcc tggtcgagct ggacggcgac gtaaacggcc acaagttcag cgtgtccggc
     6781 gagggcgagg gcgatgccac ctacggcaag ctgaccctga agttcatcag caccaccggc
     6841 aagctgcccg tgccctggcc caccctcgtg accaccctga cct
//
