LOCUS       sequence-473023-        5841 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of Grx1-RoTq-On.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5841)
  AUTHORS   Rosen PC, Glaser A, Martinez-Francois JR, Lim DC, Brooks DJ, Fu P, 
            Kim E, Kern D, Yellen G
  TITLE     Mechanism and application of thiol-disulfide redox biosensors with a
            fluorescence-lifetime readout.
  JOURNAL   Proc Natl Acad Sci U S A. 2025 May 13;122(19):e2503978122. doi: 
            10.1073/pnas.2503978122. Epub 2025 May 6.
  PUBMED    40327692
REFERENCE   2  (bases 1 to 5841)
  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: "Proc Natl 
            Acad Sci U S A."; date: "2025-05-13"; volume: "122(19):e2503978122. 
            doi"; pages: " 10.1073/pnas.2503978122. Epub 2025 May 6"
COMMENT     Sequence Label: pAAV-CAG-Grx1-RoTq-On
FEATURES             Location/Qualifiers
     source          1..5841
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     415..434
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(653..757)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(783..1238)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(920..941)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     1132..1151
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     repeat_region   1310..1439
                     /label=AAV2 ITR
     primer_bind     2303..2325
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     2387..2406
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     misc_feature    3594..4182
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3647..3667)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4065..4076)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    4225..4346
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4236..4255)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4290..4309
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     repeat_region   4508..4637
                     /label=AAV2 ITR
     rep_origin      complement(4874..5462)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4954..4973)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5633..5841,1..583,584..652))
                     /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"
ORIGIN
        1 gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag
       61 taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg
      121 tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag
      181 ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg
      241 tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc
      301 ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat
      361 tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata
      421 ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa
      481 aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca
      541 actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc
      601 aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc
      661 tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg
      721 aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac
      781 ctaaattgta agcgttaata ttttgttaaa attcgcgtta aatttttgtt aaatcagctc
      841 attttttaac caataggccg aaatcggcaa aatcccttat aaatcaaaag aatagaccga
      901 gatagggttg agtgttgttc cagtttggaa caagagtcca ctattaaaga acgtggactc
      961 caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg aaccatcacc
     1021 ctaatcaagt tttttggggt cgaggtgccg taaagcacta aatcggaacc ctaaagggag
     1081 cccccgattt agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa
     1141 agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac
     1201 cacacccgcc gcgcttaatg cgccgctaca gggcgcgtcc cattcgccat tcaggctgcg
     1261 caactgttgg gaagggcgat cggtgcgggc ctcttcgcta ttacgccagc tgcgcgctcg
     1321 ctcgctcact gaggccgccc gggcaaagcc cgggcgtcgg gcgacctttg gtcgcccggc
     1381 ctcagtgagc gagcgagcgc gcagagaggg agtggccaac tccatcacta ggggttcctt
     1441 gtagttaatg attaacccgc catgctactt atctacgtag ccatgctcta ggaagagtac
     1501 cattgacgtc aataatgacg tatgttccca tagtaacgcc aatagggact ttccattgac
     1561 gtcaatgggt ggagtattta cggtaaactg cccacttggc agtacatcaa gtgtatcata
     1621 tgccaagtac gccccctatt gacgtcaatg acggtaaatg gcccgcctgg cattatgccc
     1681 agtacatgac cttatgggac tttcctactt ggcagtacat ctacgtatta gtcatcgcta
     1741 ttaccatggt cgaggtgagc cccacgttct gcttcactct ccccatctcc cccccctccc
     1801 cacccccaat tttgtattta tttatttttt aattattttg tgcagcgatg ggggcggggg
     1861 gggggggggg cgcgccrggg gggggggggg gggggggggg gggggggggg gsgrggsggr
     1921 grggkgcggc ggcagccaat cagagcggcg cgctccgaaa gtttcctttt atggcgaggc
     1981 ggcggcggcg gcggccctat aaaaagcgaa gcgcgcggcg ggcgggagtc gctgcgcgct
     2041 gccttcgccc cgtgccccgc tccgccgccg cctcgcgccg cccgccccgg ctctgactga
     2101 ccgcgttact cccacaggtg agcgggcggg acggcccttc tcctccgggc tgtaattagc
     2161 gcttggttta atgacggctt gtttcttttc tgtggctgcg tgaaagcctt gaggggctcc
     2221 gggagggccc tttgtgcggg gggagcggct cggggctgtc cgcgggggga cggctgcctt
     2281 cgggggggac ggggcagggc ggggttcggc ttctggcgtg tgaccggcgg ctctagagcc
     2341 tctgctaacc atgttcatgc cttcttcttt ttcctacagc tcctgggcaa cgtgctggtt
     2401 attgtgctgt ctcatcattt tggcaaagaa ttggatccac catggctcaa gagtttgtga
     2461 actgcaaaat ccagcctggg aaggtggttg tgttcatcaa gcccacctgc ccgtactgca
     2521 ggagggccca agagatcctc agtcaattgc ccatcaaaca agggcttctg gaatttgtcg
     2581 atatcacagc caccaaccac actaacgaga ttcaagatta tttgcaacag ctcacgggag
     2641 caagaacggt gcctcgagtc tttattggta aagattgtat aggcggatgc agtgatctag
     2701 tctctttgca acagagtggg gaactgctga cgcggctaaa gcagattgga gctctgcaga
     2761 ctagtggtgg ttcaggtggt ggtggttcag gtggtggtgg ttcaggtgga ggaggatcag
     2821 gaggaggagg atcaggagga ggaggatcag gaggagaatt cgtgagcaaa ggcgaagaac
     2881 tgtttaccgg cgtggtgccg attctggtgg aactggatgg cgatgtgaac ggccataaat
     2941 ttagcgtgag cggcgaaggc gaaggcgatg cgacctatgg caaactgacc ctgaaattta
     3001 ttagcaccac cggcaaactg ccggtgccgt ggccgaccct ggtgaccacc ctgtcctggg
     3061 gcgtgcagtg ctttgcgcgc tatccggatc atatgaaacg ccatgatttt tttaaaagcg
     3121 cgatgccgga aggctatgtg caggaacgca ccattttttt taaagatgat ggcaactata
     3181 aaacccgcgc ggaagtgaaa tttgaaggcg ataccctggt gaaccgcatt gaactgaaag
     3241 gcattgattt taaagaagat ggcaacattc tgggccataa actggaatat aacggttggt
     3301 gcggagataa cgtgtatatt accgcggata aacagaaaaa cggcattaaa gcgaacttta
     3361 aaattcgcca taacattgaa gatggcggcg tgcagctggc ggatcattat cagcagaaca
     3421 ccccgattgg cgatggcccg gtgctgctgc cggataacca ttatctgagc acctgcagca
     3481 aactgagcaa agatccgaac gaaaaacgcg atcatatggt gtttctggaa tttgtgaccg
     3541 cggcgggcat taccctgggc atggatgaac tgtataaata aaagcttatc gataatcaac
     3601 ctctggatta caaaatttgt gaaagattga ctggtattct taactatgtt gctcctttta
     3661 cgctatgtgg atacgctgct ttaatgcctt tgtatcatgc tattgcttcc cgtatggctt
     3721 tcattttctc ctccttgtat aaatcctggt tgctgtctct ttatgaggag ttgtggcccg
     3781 ttgtcaggca acgtggcgtg gtgtgcactg tgtttgctga cgcaaccccc actggttggg
     3841 gcattgccac cacctgtcag ctcctttccg ggactttcgc tttccccctc cctattgcca
     3901 cggcggaact catcgccgcc tgccttgccc gctgctggac aggggctcgg ctgttgggca
     3961 ctgacaattc cgtggtgttg tcggggaaat catcgtcctt tccttggctg ctcgcctgtg
     4021 ttgccacctg gattctgcgc gggacgtcct tctgctacgt cccttcggcc ctcaatccag
     4081 cggaccttcc ttcccgcggc ctgctgccgg ctctgcggcc tcttccgcgt cttcgccttc
     4141 gccctcagac gagtcggatc tccctttggg ccgcctcccc gcatcgatac cgtcgacccg
     4201 ggcggccgct tcgagcagac atgataagat acattgatga gtttggacaa accacaacta
     4261 gaatgcagtg aaaaaaatgc tttatttgtg aaatttgtga tgctattgct ttatttgtaa
     4321 ccattataag ctgcaataaa caagttaaca acaacaattg cattcatttt atgtttcagg
     4381 ttcaggggga gatgtgggag gttttttaaa gcaagtaaaa cctctacaaa tgtggtaaaa
     4441 tcgataagga tcttcctaga gcatggctac gtagataagt agcatggcgg gttaatcatt
     4501 aactacaagg aacccctagt gatggagttg gccactccct ctctgcgcgc tcgctcgctc
     4561 actgaggccg ggcgaccaaa ggtcgcccga cgcccgggct ttgcccgggc ggcctcagtg
     4621 agcgagcgag cgcgcagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc
     4681 gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc
     4741 ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata
     4801 acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg
     4861 cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct
     4921 caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa
     4981 gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc
     5041 tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt
     5101 aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg
     5161 ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg
     5221 cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct
     5281 tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc
     5341 tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg
     5401 ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc
     5461 aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt
     5521 aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa
     5581 aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat
     5641 gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct
     5701 gactccccgt cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg
     5761 caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag
     5821 ccggaagggc cgagcgcaga a
//
