LOCUS       sequence-434852-        5692 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Targeting of the chemigenetic, fluorescent peroxide sensor oROS-HT 
            to actin filaments..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5692)
  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 5692)
  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-lifeact-oROS-HT
FEATURES             Location/Qualifiers
     source          1..5692
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      303..891
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     792..811
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        1066..1369
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1370..1573
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     1524..1544
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      1603..1612
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1609..1659
                     /codon_start=1
                     /product="peptide that binds filamentous actin (Riedl et 
                     al., 2008)"
                     /label=Lifeact
                     /note="first 17 amino acids of budding yeast Abp140"
                     /translation="MGVADLIKKFESISKEE"
     polyA_signal    3485..3606
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3522..3541)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3576..3595
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3613..4068)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3750..3771)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3962..3981
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4095..4199
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4201..4558
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4201..4392
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4224..4244)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4409..4544
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4593..5387
                     /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             4593..5387
                     /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(4647..4666)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5257..5276
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5575..5594)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5619..5666
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
ORIGIN
        1 tggcactctg tcgatacccc accgagaccc cattggggcc aatacgcccg cgtttcttcc
       61 ttttccccac cccacccccc aagttcgggt gaaggcccag ggctcgcagc caacgtcggg
      121 gcggcaggcc ctgccatagc ctcaggttac tcatatatac tttagattga tttaaaactt
      181 catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc
      241 ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct
      301 tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta
      361 ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc
      421 ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt aggccaccac
      481 ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct
      541 gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat
      601 aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg
      661 acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa
      721 gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg
      781 gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga
      841 cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc
      901 aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct
      961 gcgttatccc ctgattctgt ggataaccgt attaccgcca tgcattagtt attaatagta
     1021 atcaattacg gggtcattag ttcatagccc atatatggag ttccgcgtta cataacttac
     1081 ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc ccattgacgt caataatgac
     1141 gtatgttccc atagtaacgc caatagggac tttccattga cgtcaatggg tggagtattt
     1201 acggtaaact gcccacttgg cagtacatca agtgtatcat atgccaagta cgccccctat
     1261 tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc cagtacatga ccttatggga
     1321 ctttcctact tggcagtaca tctacgtatt agtcatcgct attaccatgg tgatgcggtt
     1381 ttggcagtac atcaatgggc gtggatagcg gtttgactca cggggatttc caagtctcca
     1441 ccccattgac gtcaatggga gtttgttttg gcaccaaaat caacgggact ttccaaaatg
     1501 tcgtaacaac tccgccccat tgacgcaaat gggcggtagg cgtgtacggt gggaggtcta
     1561 tataagcaga gctggtttag tgaaccgtca gatccgctag ccgccaccat gggagtcgca
     1621 gatcttataa agaaatttga atccataagt aaagaagaag gcgatccccc agtggctaca
     1681 atggagatgg cgagccagca gggcgagacg atgtccggac cgctgcacat tggtttgatt
     1741 cccacagttg gaccgtacct gctaccgcat attatcccta tgctgcacca gacctttcca
     1801 aagctggaaa tgtatctgca tgaagcacag acccaccagt tactggcgca actggacagc
     1861 ggcaaactcg attgcgtgat cctcgcactg gtgaaagaga gcgaagcatt cattgaagtg
     1921 ccgttgtttg atgagccaat gttgctggct atctatgaag atcacccgtg ggcgaaccgc
     1981 gaatgcgtac cgatggccga tctggcaggg gaaaaactgc tgatgctgga agatggtcac
     2041 tgtttgcgcg atcaggcaat gggtttctgt tttgaagccg gggcgctgag ctcaacgggt
     2101 gagaccttcc aggccttccg caccaccgac gtcggccgca agctgatcat cgatcagaac
     2161 gtttttatcg agggtacgct gccgatgggt gtcgtccgcc cgctgactga agtcgagatg
     2221 gaccattacc gcgagccgtt cctgaatcct gttgaccgcg agccactgtg gcgcttccca
     2281 aacgagctgc caatcgccgg tgagccagcg aacatcgtcg cgctggtcga agaatacatg
     2341 gactggctgc accagtcccc tgtcccgaag ctgctgttct ggggcacccc aggcgttctg
     2401 atcccaccgg ccgaagccgc tcgcctggcc aaaagcctgc ctaactgcaa ggctgtggac
     2461 atcggcccgg gtctgaatct gctgcaagaa gacaacccgg acctgatcgg cagcgagatc
     2521 gcgcgctggc tgtcgacgct cgagatttcc ggcggaggaa caggtggttc tggtggaaca
     2581 gggggtagcg gaggtacagg aggaagtatg gcggagatcg gaactggatt cccgtttgat
     2641 ccgcattatg tggaagttct gggagagcgc atgcattatg tggacgttgg tcctcgtgat
     2701 gggacaccag tgctgttcct tcacggcaat ccgacatcgt cgtacgtgtg gcgtaatatc
     2761 atcccgcacg ttgcccccac gcaccgctgc attgcccctg acttaattgg tatggggaaa
     2821 agtgataagc ctgatctggg gtacttcttt gacgaccacg tacgcttcat ggatgctttt
     2881 attgaagcat tgggtttgga ggaagtagtt ttggtgatcc atgattgggg tagtgctctg
     2941 gggttccatt gggccaagcg taacccagaa cgcgtgaaag gaattgcctt tatggagttc
     3001 atccgtccga ttccaacatg ggacgaatgg cgaccggacc aactggatga agatacacac
     3061 ttccgcgcga ccagcctgga aactctgcgc aacatggtgg cggcaggtag cgggatcact
     3121 ttactgccag cgctggctgt gccgccggag cgcaaacgcg atggggttgt ttatctgccg
     3181 tgcattaagc cggaaccacg ccgcactatt ggtctggttt atcgtcctgg ctcaccgctg
     3241 cgcagccgct atgagcagct ggcagaggcc atccgcgcaa gaatggatgg ccatttcgat
     3301 aaagttttaa aacaggcggt ttaagaattc tgcagtcgac ggtaccgcgg gcccgggatc
     3361 caccggatct agataactga tcataatcag ccataccaca tttgtagagg ttttacttgc
     3421 tttaaaaaac ctcccacacc tccccctgaa cctgaaacat aaaatgaatg caattgttgt
     3481 tgttaacttg tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt
     3541 cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt
     3601 atcttaaggc gtaaattgta agcgttaata ttttgttaaa attcgcgtta aatttttgtt
     3661 aaatcagctc attttttaac caataggccg aaatcggcaa aatcccttat aaatcaaaag
     3721 aatagaccga gatagggttg agtgttgttc cagtttggaa caagagtcca ctattaaaga
     3781 acgtggactc caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg
     3841 aaccatcacc ctaatcaagt tttttggggt cgaggtgccg taaagcacta aatcggaacc
     3901 ctaaagggag cccccgattt agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg
     3961 aagggaagaa agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc
     4021 gcgtaaccac cacacccgcc gcgcttaatg cgccgctaca gggcgcgtca ggtggcactt
     4081 ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt
     4141 atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagtc
     4201 ctgaggcgga aagaaccagc tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg
     4261 ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg
     4321 aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc
     4381 aaccatagtc ccgcccctaa ctccgcccat cccgccccta actccgccca gttccgccca
     4441 ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg ccgcctcggc
     4501 ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctaggct tttgcaaaga
     4561 tcgatcaaga gacaggatga ggatcgtttc gcatgattga acaagatgga ttgcacgcag
     4621 gttctccggc cgcttgggtg gagaggctat tcggctatga ctgggcacaa cagacaatcg
     4681 gctgctctga tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca
     4741 agaccgacct gtccggtgcc ctgaatgaac tgcaagacga ggcagcgcgg ctatcgtggc
     4801 tggccacgac gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg
     4861 actggctgct attgggcgaa gtgccggggc aggatctcct gtcatctcac cttgctcctg
     4921 ccgagaaagt atccatcatg gctgatgcaa tgcggcggct gcatacgctt gatccggcta
     4981 cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact cggatggaag
     5041 ccggtcttgt cgatcaggat gatctggacg aagagcatca ggggctcgcg ccagccgaac
     5101 tgttcgccag gctcaaggcg agcatgcccg acggcgagga tctcgtcgtg acccatggcg
     5161 atgcctgctt gccgaatatc atggtggaaa atggccgctt ttctggattc atcgactgtg
     5221 gccggctggg tgtggcggac cgctatcagg acatagcgtt ggctacccgt gatattgctg
     5281 aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc gccgctcccg
     5341 attcgcagcg catcgccttc tatcgccttc ttgacgagtt cttctgagcg ggactctggg
     5401 gttcgaaatg accgaccaag cgacgcccaa cctgccatca cgagatttcg attccaccgc
     5461 cgccttctat gaaaggttgg gcttcggaat cgttttccgg gacgccggct ggatgatcct
     5521 ccagcgcggg gatctcatgc tggagttctt cgcccaccct agggggaggc taactgaaac
     5581 acggaaggag acaataccgg aaggaacccg cgctatgacg gcaataaaaa gacagaataa
     5641 aacgcacggt gttgggtcgt ttgttcataa acgcggggtt cggtcccagg gc
//
