LOCUS       sequence-429715-        6138 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Encodes the genetically encoded, green fluorescent peroxide sensor 
            oROS-G in AAV viral vectors..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6138)
  AUTHORS   Lee J.D., Won. W., Kimball K., Wang Y., Yeboah F., Evitts K.M., 
            Neiswanger C., Schattauer S., Rappleye M., Bremner S.B., Chun C., 
            Smith N., Mack D.L., Young J.E., Lee C.J., Chavkin C., Berndt A.
  TITLE     Structure-guided engineering of a fast genetically encoded sensor 
            for real-time H2O2 monitoring
  JOURNAL   bioRxiv 2024.01.31.578117
REFERENCE   2  (bases 1 to 6138)
  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 
            2024.01.31.578117"
COMMENT     Sequence Label: AAV2_CAG_oROS-G
FEATURES             Location/Qualifiers
     source          1..6138
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..130
                     /label=AAV2 ITR
     primer_bind     997..1019
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1081..1100
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     regulatory      1133..1142
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     1753..1774
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1880..1901)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2141..2160)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     misc_feature    2582..3170
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2635..2655)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3053..3064)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3213..3334
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3224..3243)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3278..3297
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     repeat_region   3496..3625
                     /label=AAV2 ITR
     rep_origin      complement(3862..4450)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3942..3961)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4621..5412,5413..5481))
                     /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     5244..5263
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5482..5586)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(5612..6067)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5749..5770)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5961..5980
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 ctgcgcgctc gctcgctcac tgaggccgcc cgggcaaagc ccgggcgtcg ggcgaccttt
       61 ggtcgcccgg cctcagtgag cgagcgagcg cgcagagagg gagtggccaa ctccatcact
      121 aggggttcct tgtagttaat gattaacccg ccatgctact tatctacgta gccatgctct
      181 aggaagagta ccattgacgt caataatgac gtatgttccc atagtaacgc caatagggac
      241 tttccattga cgtcaatggg tggagtattt acggtaaact gcccacttgg cagtacatca
      301 agtgtatcat atgccaagta cgccccctat tgacgtcaat gacggtaaat ggcccgcctg
      361 gcattatgcc cagtacatga ccttatggga ctttcctact tggcagtaca tctacgtatt
      421 agtcatcgct attaccatgg tcgaggtgag ccccacgttc tgcttcactc tccccatctc
      481 ccccccctcc ccacccccaa ttttgtattt atttattttt taattatttt gtgcagcgat
      541 gggggcgggg gggggggggg ggcgcgcgcc rggggggggg gggggggggg gggggggggg
      601 sggggsgrgg cggrgrggkg cggcggcagc caatcagagc ggcgcgctcc gaaagtttcc
      661 ttttatggcg aggcggcggc ggcggcggcc ctataaaaag cgaagcgcgc ggcgggcggg
      721 agtcgctgcg cgctgccttc gccccgtgcc ccgctccgcc gccgcctcgc gccgcccgcc
      781 ccggctctga ctgaccgcgt tactcccaca ggtgagcggg cgggacggcc cttctcctcc
      841 gggctgtaat tagcgcttgg tttaatgacg gcttgtttct tttctgtggc tgcgtgaaag
      901 ccttgagggg ctccgggagg gccctttgtg cggggggagc ggctcggggc tgtccgcggg
      961 gggacggctg ccttcggggg ggacggggca gggcggggtt cggcttctgg cgtgtgaccg
     1021 gcggctctag agcctctgct aaccatgttc atgccttctt ctttttccta cagctcctgg
     1081 gcaacgtgct ggttattgtg ctgtctcatc attttggcaa agaattggat ccgccaccat
     1141 ggagatggcg agccagcagg gcgagacgat gtccggaccg ctgcacattg gtttgattcc
     1201 cacagttgga ccgtacctgc taccgcatat tatccctatg ctgcaccaga cctttccaaa
     1261 gctggaaatg tatctgcatg aagcacagac ccaccagtta ctggcgcaac tggacagcgg
     1321 caaactcgat tgcgtgatcc tcgcactggt gaaagagagc gaagcattca ttgaagtgcc
     1381 gttgtttgat gagccaatgt tgctggctat ctatgaagat cacccgtggg cgaaccgcga
     1441 atgcgtaccg atggccgatc tggcagggga aaaactgctg atgctggaag atggtcactg
     1501 tttgcgcgat caggcaatgg gtttctgttt tgaagcctca ctcgagaacg tctatatcaa
     1561 ggccgacaag cagaagaacg gcatcaaggc gaacttcaag atccgccaca acatcgagga
     1621 cggcggcgtg cagctcgcct accactacca gcagaacacc cccatcggcg acggccccgt
     1681 gctgctgccc gacaaccact acctgagcgt gcagtccaaa ctttcgaaag accccaacga
     1741 gaagcgcgat cacatggtcc tgctggagtt cgtgaccgcc gccgggatca ctctcggcat
     1801 ggacgagctg tacaagggcg gtaccggagg gagcatggtg agcaagggcg aggagctgtt
     1861 caccggggtg gtgcccatcc tggtcgagct ggacggcgac gtaaacggcc acaagttcag
     1921 cgtgtccggc gagggtgagg gcgatgccac ctacggcaag ctgaccctga agttcatctg
     1981 caccaccggc aagctgcccg tgccctggcc caccctcgtg accaccctga cctacggcgt
     2041 gcagtgcttc agccgctacc ccgaccacat gaagcagcac gacttcttca agtccgccat
     2101 gcccgaaggc tacatccagg agcgcaccat cttcttcaag gacgacggca actacaagac
     2161 ccgcgccgag gtgaagttcg agggcgacac cctggtgaac cgcatcgagc tgaagggcat
     2221 cgacttcaag gaggacggca acatcctggg gcacaagctg gagtacaacc tgccggacca
     2281 actgggggcg gattacgata cacacttccg cgcgaccagc ctggaaactc tgcgcaacat
     2341 ggtggcggca ggtagcggga tcactttact gccagcgctg gctgtgccgc cggagcgcaa
     2401 acgcgatggg gttgtttatc tgccgtgcat taagccggaa ccacgccgca ctattggtct
     2461 ggtttatcgt cctggctcac cgctgcgcag ccgctatgag cagctggcag aggccatccg
     2521 cgcaagaatg gatggccatt tcgataaagt tttaaaacag gcggtttaaa agcttatcga
     2581 taatcaacct ctggattaca aaatttgtga aagattgact ggtattctta actatgttgc
     2641 tccttttacg ctatgtggat acgctgcttt aatgcctttg tatcatgcta ttgcttcccg
     2701 tatggctttc attttctcct ccttgtataa atcctggttg ctgtctcttt atgaggagtt
     2761 gtggcccgtt gtcaggcaac gtggcgtggt gtgcactgtg tttgctgacg caacccccac
     2821 tggttggggc attgccacca cctgtcagct cctttccggg actttcgctt tccccctccc
     2881 tattgccacg gcggaactca tcgccgcctg ccttgcccgc tgctggacag gggctcggct
     2941 gttgggcact gacaattccg tggtgttgtc ggggaaatca tcgtcctttc cttggctgct
     3001 cgcctgtgtt gccacctgga ttctgcgcgg gacgtccttc tgctacgtcc cttcggccct
     3061 caatccagcg gaccttcctt cccgcggcct gctgccggct ctgcggcctc ttccgcgtct
     3121 tcgccttcgc cctcagacga gtcggatctc cctttgggcc gcctccccgc atcgataccg
     3181 tcgacccggg cggccgcttc gagcagacat gataagatac attgatgagt ttggacaaac
     3241 cacaactaga atgcagtgaa aaaaatgctt tatttgtgaa atttgtgatg ctattgcttt
     3301 atttgtaacc attataagct gcaataaaca agttaacaac aacaattgca ttcattttat
     3361 gtttcaggtt cagggggaga tgtgggaggt tttttaaagc aagtaaaacc tctacaaatg
     3421 tggtaaaatc gataaggatc ttcctagagc atggctacgt agataagtag catggcgggt
     3481 taatcattaa ctacaaggaa cccctagtga tggagttggc cactccctct ctgcgcgctc
     3541 gctcgctcac tgaggccggg cgaccaaagg tcgcccgacg cccgggcttt gcccgggcgg
     3601 cctcagtgag cgagcgagcg cgcagctgca ttaatgaatc ggccaacgcg cggggagagg
     3661 cggtttgcgt attkggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt
     3721 tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc
     3781 aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa
     3841 aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa
     3901 tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc
     3961 ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc
     4021 cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag
     4081 ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga
     4141 ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc
     4201 gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac
     4261 agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg
     4321 cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca
     4381 aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa
     4441 aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa
     4501 ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt
     4561 aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag
     4621 ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat
     4681 agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc
     4741 cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa
     4801 ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca
     4861 gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa
     4921 cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt
     4981 cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc
     5041 ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact
     5101 catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc
     5161 tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg
     5221 ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct
     5281 catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc
     5341 cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag
     5401 cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac
     5461 acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg
     5521 ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt
     5581 tccgcgcaca tttccccgaa aagtgccacc taaattgtaa gcgttaatat tttgttaaaa
     5641 ttcgcgttaa atttttgtta aatcagctca ttttttaacc aataggccga aatcggcaaa
     5701 atcccttata aatcaaaaga atagaccgag atagggttga gtgttgttcc agtttggaac
     5761 aagagtccac tattaaagaa cgtggactcc aacgtcaaag ggcgaaaaac cgtctatcag
     5821 ggcgatggcc cactacgtga accatcaccc taatcaagtt ttttggggtc gaggtgccgt
     5881 aaagcactaa atcggaaccc taaagggagc ccccgattta gagcttgacg gggaaagccg
     5941 gcgaacgtgg cgagaaagga agggaagaaa gcgaaaggag cgggcgctag ggcgctggca
     6001 agtgtagcgg tcacgctgcg cgtaaccacc acacccgccg cgcttaatgc gccgctacag
     6061 ggcgcgtccc attcgccatt caggctgcgc aactgttggg aagggcgatc ggtgcgggcc
     6121 tcttcgctat tacgccag
//
