LOCUS       sequence-435272-        5623 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses the loss-of-function mutation C199S of the genetically 
            encoded, chemigenetic hydrogen peroxide sensor oROS-HT in mamalian 
            cells..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5623)
  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 5623)
  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: pC1_oROS-HT_LF(C199S)
FEATURES             Location/Qualifiers
     source          1..5623
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      3..12
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     polyA_signal    1816..1937
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1853..1872)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1907..1926
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(1944..2399)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2081..2102)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2293..2312
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2426..2530
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2532..2889
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2532..2723
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2555..2575)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      2740..2875
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             2924..3718
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             2924..3718
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(2978..2997)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3588..3607
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(3906..3925)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    3950..3997
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4326..4914
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4815..4834
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        5089..5392
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        5393..5596
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     5547..5567
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
ORIGIN
        1 ccgccaccat ggagatggcg agccagcagg gcgagacgat gtccggaccg ctgcacattg
       61 gtttgattcc cacagttgga ccgtacctgc taccgcatat tatccctatg ctgcaccaga
      121 cctttccaaa gctggaaatg tatctgcatg aagcacagac ccaccagtta ctggcgcaac
      181 tggacagcgg caaactcgat tgcgtgatcc tcgcactggt gaaagagagc gaagcattca
      241 ttgaagtgcc gttgtttgat gagccaatgt tgctggctat ctatgaagat cacccgtggg
      301 cgaaccgcga atgcgtaccg atggccgatc tggcagggga aaaactgctg atgctggaag
      361 atggtcacag tttgcgcgat caggcaatgg gtttctgttt tgaagccggg gcgctgagct
      421 caacgggtga gaccttccag gccttccgca ccaccgacgt cggccgcaag ctgatcatcg
      481 atcagaacgt ttttatcgag ggtacgctgc cgatgggtgt cgtccgcccg ctgactgaag
      541 tcgagatgga ccattaccgc gagccgttcc tgaatcctgt tgaccgcgag ccactgtggc
      601 gcttcccaaa cgagctgcca atcgccggtg agccagcgaa catcgtcgcg ctggtcgaag
      661 aatacatgga ctggctgcac cagtcccctg tcccgaagct gctgttctgg ggcaccccag
      721 gcgttctgat cccaccggcc gaagccgctc gcctggccaa aagcctgcct aactgcaagg
      781 ctgtggacat cggcccgggt ctgaatctgc tgcaagaaga caacccggac ctgatcggca
      841 gcgagatcgc gcgctggctg tcgacgctcg agatttccgg cggaggaaca ggtggttctg
      901 gtggaacagg gggtagcgga ggtacaggag gaagtatggc ggagatcgga actggattcc
      961 cgtttgatcc gcattatgtg gaagttctgg gagagcgcat gcattatgtg gacgttggtc
     1021 ctcgtgatgg gacaccagtg ctgttccttc acggcaatcc gacatcgtcg tacgtgtggc
     1081 gtaatatcat cccgcacgtt gcccccacgc accgctgcat tgcccctgac ttaattggta
     1141 tggggaaaag tgataagcct gatctggggt acttctttga cgaccacgta cgcttcatgg
     1201 atgcttttat tgaagcattg ggtttggagg aagtagtttt ggtgatccat gattggggta
     1261 gtgctctggg gttccattgg gccaagcgta acccagaacg cgtgaaagga attgccttta
     1321 tggagttcat ccgtccgatt ccaacatggg acgaatggcg accggaccaa ctggatgaag
     1381 atacacactt ccgcgcgacc agcctggaaa ctctgcgcaa catggtggcg gcaggtagcg
     1441 ggatcacttt actgccagcg ctggctgtgc cgccggagcg caaacgcgat ggggttgttt
     1501 atctgccgtg cattaagccg gaaccacgcc gcactattgg tctggtttat cgtcctggct
     1561 caccgctgcg cagccgctat gagcagctgg cagaggccat ccgcgcaaga atggatggcc
     1621 atttcgataa agttttaaaa caggcggttt aatgagaatt ctgcagtcga cggtaccgcg
     1681 ggcccgggat ccaccggatc tagataactg atcataatca gccataccac atttgtagag
     1741 gttttacttg ctttaaaaaa cctcccacac ctccccctga acctgaaaca taaaatgaat
     1801 gcaattgttg ttgttaactt gtttattgca gcttataatg gttacaaata aagcaatagc
     1861 atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa
     1921 ctcatcaatg tatcttaagg cgtaaattgt aagcgttaat attttgttaa aattcgcgtt
     1981 aaatttttgt taaatcagct cattttttaa ccaataggcc gaaatcggca aaatccctta
     2041 taaatcaaaa gaatagaccg agatagggtt gagtgttgtt ccagtttgga acaagagtcc
     2101 actattaaag aacgtggact ccaacgtcaa agggcgaaaa accgtctatc agggcgatgg
     2161 cccactacgt gaaccatcac cctaatcaag ttttttgggg tcgaggtgcc gtaaagcact
     2221 aaatcggaac cctaaaggga gcccccgatt tagagcttga cggggaaagc cggcgaacgt
     2281 ggcgagaaag gaagggaaga aagcgaaagg agcgggcgct agggcgctgg caagtgtagc
     2341 ggtcacgctg cgcgtaacca ccacacccgc cgcgcttaat gcgccgctac agggcgcgtc
     2401 aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca
     2461 ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa
     2521 aaggaagagt cctgaggcgg aaagaaccag ctgtggaatg tgtgtcagtt agggtgtgga
     2581 aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca
     2641 accaggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc
     2701 aattagtcag caaccatagt cccgccccta actccgccca tcccgcccct aactccgccc
     2761 agttccgccc attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag
     2821 gccgcctcgg cctctgagct attccagaag tagtgaggag gcttttttgg aggcctaggc
     2881 ttttgcaaag atcgatcaag agacaggatg aggatcgttt cgcatgattg aacaagatgg
     2941 attgcacgca ggttctccgg ccgcttgggt ggagaggcta ttcggctatg actgggcaca
     3001 acagacaatc ggctgctctg atgccgccgt gttccggctg tcagcgcagg ggcgcccggt
     3061 tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa ctgcaagacg aggcagcgcg
     3121 gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct gtgctcgacg ttgtcactga
     3181 agcgggaagg gactggctgc tattgggcga agtgccgggg caggatctcc tgtcatctca
     3241 ccttgctcct gccgagaaag tatccatcat ggctgatgca atgcggcggc tgcatacgct
     3301 tgatccggct acctgcccat tcgaccacca agcgaaacat cgcatcgagc gagcacgtac
     3361 tcggatggaa gccggtcttg tcgatcagga tgatctggac gaagagcatc aggggctcgc
     3421 gccagccgaa ctgttcgcca ggctcaaggc gagcatgccc gacggcgagg atctcgtcgt
     3481 gacccatggc gatgcctgct tgccgaatat catggtggaa aatggccgct tttctggatt
     3541 catcgactgt ggccggctgg gtgtggcgga ccgctatcag gacatagcgt tggctacccg
     3601 tgatattgct gaagagcttg gcggcgaatg ggctgaccgc ttcctcgtgc tttacggtat
     3661 cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt cttgacgagt tcttctgagc
     3721 gggactctgg ggttcgaaat gaccgaccaa gcgacgccca acctgccatc acgagatttc
     3781 gattccaccg ccgccttcta tgaaaggttg ggcttcggaa tcgttttccg ggacgccggc
     3841 tggatgatcc tccagcgcgg ggatctcatg ctggagttct tcgcccaccc tagggggagg
     3901 ctaactgaaa cacggaagga gacaataccg gaaggaaccc gcgctatgac ggcaataaaa
     3961 agacagaata aaacgcacgg tgttgggtcg tttgttcata aacgcggggt tcggtcccag
     4021 ggctggcact ctgtcgatac cccaccgaga ccccattggg gccaatacgc ccgcgtttct
     4081 tccttttccc caccccaccc cccaagttcg ggtgaaggcc cagggctcgc agccaacgtc
     4141 ggggcggcag gccctgccat agcctcaggt tactcatata tactttagat tgatttaaaa
     4201 cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa
     4261 atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga
     4321 tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg
     4381 ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact
     4441 ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac
     4501 cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg
     4561 gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg
     4621 gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga
     4681 acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc
     4741 gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg
     4801 agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc
     4861 tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc
     4921 agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt
     4981 cctgcgttat cccctgattc tgtggataac cgtattaccg ccatgcatta gttattaata
     5041 gtaatcaatt acggggtcat tagttcatag cccatatatg gagttccgcg ttacataact
     5101 tacggtaaat ggcccgcctg gctgaccgcc caacgacccc cgcccattga cgtcaataat
     5161 gacgtatgtt cccatagtaa cgccaatagg gactttccat tgacgtcaat gggtggagta
     5221 tttacggtaa actgcccact tggcagtaca tcaagtgtat catatgccaa gtacgccccc
     5281 tattgacgtc aatgacggta aatggcccgc ctggcattat gcccagtaca tgaccttatg
     5341 ggactttcct acttggcagt acatctacgt attagtcatc gctattacca tggtgatgcg
     5401 gttttggcag tacatcaatg ggcgtggata gcggtttgac tcacggggat ttccaagtct
     5461 ccaccccatt gacgtcaatg ggagtttgtt ttggcaccaa aatcaacggg actttccaaa
     5521 atgtcgtaac aactccgccc cattgacgca aatgggcggt aggcgtgtac ggtgggaggt
     5581 ctatataagc agagctggtt tagtgaaccg tcagatccgc tag
//
