LOCUS       sequence-429726-        6351 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Encodes the loss-of-function mutation C199S of genetically encoded, 
            chemigenetic fluorescent peroxide sensor oROS-HT in AAV viral 
            vectors.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6351)
  AUTHORS   Lee J.D., Nguyen A., Jin Z.R., Moghadasi A., Gibbs C.E., Wait S.J., 
            Evitts K.M., Asencio A., Bremner S.B., Zuniga S., Chavan V., 
            Williams A., Smith N., Regnier M., Young J.E., Mack D., Nance E., 
            Boyle P.M., Berndt A.
  TITLE     Far-red and sensitive sensor for monitoring real time H2O2 dynamics 
            with subcellular resolution and in multi-parametric imaging 
            applications
  JOURNAL   bioRxiv 2024.02.06.579232
REFERENCE   2  (bases 1 to 6351)
  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: "bioRxiv 
            2024.02.06.579232"
COMMENT     Sequence Label: AAV2_CAG_oROS-HT(C199S)_WPRE
FEATURES             Location/Qualifiers
     source          1..6351
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      1..10
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     misc_feature    1663..2251
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(1716..1736)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2134..2145)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    2294..2415
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2305..2324)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2359..2378
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     repeat_region   2577..2706
                     /label=AAV2 ITR
     rep_origin      complement(2943..3531)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3023..3042)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3702..4493,4494..4562))
                     /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     4325..4344
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4563..4667)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(4693..5148)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4830..4851)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5042..5061
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     repeat_region   5220..5349
                     /label=AAV2 ITR
     primer_bind     6216..6238
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     6300..6319
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
ORIGIN
        1 gccaccatgg agatggcgag ccagcagggc gagacgatgt ccggaccgct gcacattggt
       61 ttgattccca cagttggacc gtacctgcta ccgcatatta tccctatgct gcaccagacc
      121 tttccaaagc tggaaatgta tctgcatgaa gcacagaccc accagttact ggcgcaactg
      181 gacagcggca aactcgattg cgtgatcctc gcactggtga aagagagcga agcattcatt
      241 gaagtgccgt tgtttgatga gccaatgttg ctggctatct atgaagatca cccgtgggcg
      301 aaccgcgaat gcgtaccgat ggccgatctg gcaggggaaa aactgctgat gctggaagat
      361 ggtcacagtt tgcgcgatca ggcaatgggt ttctgttttg aagccggggc gctgagctca
      421 acgggtgaga ccttccaggc cttccgcacc accgacgtcg gccgcaagct gatcatcgat
      481 cagaacgttt ttatcgaggg tacgctgccg atgggtgtcg tccgcccgct gactgaagtc
      541 gagatggacc attaccgcga gccgttcctg aatcctgttg accgcgagcc actgtggcgc
      601 ttcccaaacg agctgccaat cgccggtgag ccagcgaaca tcgtcgcgct ggtcgaagaa
      661 tacatggact ggctgcacca gtcccctgtc ccgaagctgc tgttctgggg caccccaggc
      721 gttctgatcc caccggccga agccgctcgc ctggccaaaa gcctgcctaa ctgcaaggct
      781 gtggacatcg gcccgggtct gaatctgctg caagaagaca acccggacct gatcggcagc
      841 gagatcgcgc gctggctgtc gacgctcgag atttccggcg gaggaacagg tggttctggt
      901 ggaacagggg gtagcggagg tacaggagga agtatggcgg agatcggaac tggattcccg
      961 tttgatccgc attatgtgga agttctggga gagcgcatgc attatgtgga cgttggtcct
     1021 cgtgatggga caccagtgct gttccttcac ggcaatccga catcgtcgta cgtgtggcgt
     1081 aatatcatcc cgcacgttgc ccccacgcac cgctgcattg cccctgactt aattggtatg
     1141 gggaaaagtg ataagcctga tctggggtac ttctttgacg accacgtacg cttcatggat
     1201 gcttttattg aagcattggg tttggaggaa gtagttttgg tgatccatga ttggggtagt
     1261 gctctggggt tccattgggc caagcgtaac ccagaacgcg tgaaaggaat tgcctttatg
     1321 gagttcatcc gtccgattcc aacatgggac gaatggcgac cggaccaact ggatgaagat
     1381 acacacttcc gcgcgaccag cctggaaact ctgcgcaaca tggtggcggc aggtagcggg
     1441 atcactttac tgccagcgct ggctgtgccg ccggagcgca aacgcgatgg ggttgtttat
     1501 ctgccgtgca ttaagccgga accacgccgc actattggtc tggtttatcg tcctggctca
     1561 ccgctgcgca gccgctatga gcagctggca gaggccatcc gcgcaagaat ggatggccat
     1621 ttcgataaag ttttaaaaca ggcggtttaa aagcttatcg ataatcaacc tctggattac
     1681 aaaatttgtg aaagattgac tggtattctt aactatgttg ctccttttac gctatgtgga
     1741 tacgctgctt taatgccttt gtatcatgct attgcttccc gtatggcttt cattttctcc
     1801 tccttgtata aatcctggtt gctgtctctt tatgaggagt tgtggcccgt tgtcaggcaa
     1861 cgtggcgtgg tgtgcactgt gtttgctgac gcaaccccca ctggttgggg cattgccacc
     1921 acctgtcagc tcctttccgg gactttcgct ttccccctcc ctattgccac ggcggaactc
     1981 atcgccgcct gccttgcccg ctgctggaca ggggctcggc tgttgggcac tgacaattcc
     2041 gtggtgttgt cggggaaatc atcgtccttt ccttggctgc tcgcctgtgt tgccacctgg
     2101 attctgcgcg ggacgtcctt ctgctacgtc ccttcggccc tcaatccagc ggaccttcct
     2161 tcccgcggcc tgctgccggc tctgcggcct cttccgcgtc ttcgccttcg ccctcagacg
     2221 agtcggatct ccctttgggc cgcctccccg catcgatacc gtcgacccgg gcggccgctt
     2281 cgagcagaca tgataagata cattgatgag tttggacaaa ccacaactag aatgcagtga
     2341 aaaaaatgct ttatttgtga aatttgtgat gctattgctt tatttgtaac cattataagc
     2401 tgcaataaac aagttaacaa caacaattgc attcatttta tgtttcaggt tcagggggag
     2461 atgtgggagg ttttttaaag caagtaaaac ctctacaaat gtggtaaaat cgataaggat
     2521 cttcctagag catggctacg tagataagta gcatggcggg ttaatcatta actacaagga
     2581 acccctagtg atggagttgg ccactccctc tctgcgcgct cgctcgctca ctgaggccgg
     2641 gcgaccaaag gtcgcccgac gcccgggctt tgcccgggcg gcctcagtga gcgagcgagc
     2701 gcgcagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg tattkggcgc
     2761 tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta
     2821 tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag
     2881 aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg
     2941 tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg
     3001 tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg
     3061 cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga
     3121 agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc
     3181 tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt
     3241 aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact
     3301 ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg
     3361 cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt
     3421 accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt
     3481 ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct
     3541 ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg
     3601 gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt
     3661 aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt
     3721 gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc
     3781 gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg
     3841 cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc
     3901 gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg
     3961 gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca
     4021 ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga
     4081 tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct
     4141 ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg
     4201 cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca
     4261 accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata
     4321 cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct
     4381 tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact
     4441 cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa
     4501 acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc
     4561 atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga
     4621 tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga
     4681 aaagtgccac ctaaattgta agcgttaata ttttgttaaa attcgcgtta aatttttgtt
     4741 aaatcagctc attttttaac caataggccg aaatcggcaa aatcccttat aaatcaaaag
     4801 aatagaccga gatagggttg agtgttgttc cagtttggaa caagagtcca ctattaaaga
     4861 acgtggactc caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg
     4921 aaccatcacc ctaatcaagt tttttggggt cgaggtgccg taaagcacta aatcggaacc
     4981 ctaaagggag cccccgattt agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg
     5041 aagggaagaa agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc
     5101 gcgtaaccac cacacccgcc gcgcttaatg cgccgctaca gggcgcgtcc cattcgccat
     5161 tcaggctgcg caactgttgg gaagggcgat cggtgcgggc ctcttcgcta ttacgccagc
     5221 tgcgcgctcg ctcgctcact gaggccgccc gggcaaagcc cgggcgtcgg gcgacctttg
     5281 gtcgcccggc ctcagtgagc gagcgagcgc gcagagaggg agtggccaac tccatcacta
     5341 ggggttcctt gtagttaatg attaacccgc catgctactt atctacgtag ccatgctcta
     5401 ggaagagtac cattgacgtc aataatgacg tatgttccca tagtaacgcc aatagggact
     5461 ttccattgac gtcaatgggt ggagtattta cggtaaactg cccacttggc agtacatcaa
     5521 gtgtatcata tgccaagtac gccccctatt gacgtcaatg acggtaaatg gcccgcctgg
     5581 cattatgccc agtacatgac cttatgggac tttcctactt ggcagtacat ctacgtatta
     5641 gtcatcgcta ttaccatggt cgaggtgagc cccacgttct gcttcactct ccccatctcc
     5701 cccccctccc cacccccaat tttgtattta tttatttttt aattattttg tgcagcgatg
     5761 ggggcggggg gggggggggg gcgcgcgccr gggggggsgg gggggggsgg ggggsggggs
     5821 ggggsgrggs ggrgrggkgc ggcggcagcc aatcagagcg gcgcgctccg aaagtttcct
     5881 tttatggcga ggcggcggcg gcggcggccc tataaaaagc gaagcgcgcg gcgggcggga
     5941 gtcgctgcgc gctgccttcg ccccgtgccc cgctccgccg ccgcctcgcg ccgcccgccc
     6001 cggctctgac tgaccgcgtt actcccacag gtgagcgggc gggacggccc ttctcctccg
     6061 ggctgtaatt agcgcttggt ttaatgacgg cttgtttctt ttctgtggct gcgtgaaagc
     6121 cttgaggggc tccgggaggg ccctttgtgc ggggggagcg gctcggggct gtccgcgggg
     6181 ggacggctgc cttcgggggg gacggggcag ggcggggttc ggcttctggc gtgtgaccgg
     6241 cggctctaga gcctctgcta accatgttca tgccttcttc tttttcctac agctcctggg
     6301 caacgtgctg gttattgtgc tgtctcatca ttttggcaaa gaattggatc c
//
