LOCUS       sequence-430079-        6918 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Targeting of the chemigenetic, fluorescent peroxide sensor oROS-HT 
            to the extracellular site of cell membranes..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6918)
  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 6918)
  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: pDisplay-oROS-HT
FEATURES             Location/Qualifiers
     source          1..6918
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             1..30
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             34..180
                     /codon_start=1
                     /product="transmembrane domain from platelet derived growth
                     factor receptor beta"
                     /label=PDGFR-beta TM domain
                     /translation="AVGQDTQEVIVVPHSLPFKVVVISAILALVVLTIISLIILIMLWQ
                     KKPR"
     primer_bind     complement(202..219)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    208..432
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      complement(564..1152)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(644..663)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    1481..1528
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     primer_bind     1553..1572
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     CDS             complement(1760..2554)
                     /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             complement(1760..2554)
                     /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(1871..1890)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     2481..2500
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     complement(2653..2672)
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     enhancer        2751..2942
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     2899..2919
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     CDS             complement(join(3002..3793,3794..3862))
                     /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     3625..3644
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3863..3967)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      3994..4449
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4081..4100)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4291..4312
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     enhancer        4461..4840
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4841..5044
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     4995..5015
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     5089..5108
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        5089..5107
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     sig_peptide     5188..5247
                     /codon_start=1
                     /product="leader sequence from mouse immunoglobulin kappa 
                     light chain"
                     /label=Ig-kappa leader
                     /translation="METDTLLLWVLLLWVPGSTG"
     CDS             5251..5277
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
ORIGIN
        1 gaacaaaaac tcatctcaga agaggatctg aatgctgtgg gccaggacac gcaggaggtc
       61 atcgtggtgc cacactcctt gccctttaag gtggtggtga tctcagccat cctggccctg
      121 gtggtgctca ccatcatctc ccttatcatc ctcatcatgc tttggcagaa gaagccacgt
      181 taggcggccg ctcgagatca gcctcgactg tgccttctag ttgccagcca tctgttgttt
      241 gcccctcccc cgtgccttcc ttgaccctgg aaggtgccac tcccactgtc ctttcctaat
      301 aaaatgagga aattgcatcg cattgtctga gtaggtgtca ttctattctg gggggtgggg
      361 tgaggcagga cagcaagggg gaggattggg aagacaatag caggcatgct ggggatgcgg
      421 tgggctctat ggcttctgag gcggaaagaa ccagtggcgg taatacggtt atccacagaa
      481 tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt
      541 aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa
      601 aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt
      661 ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg
      721 tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc
      781 agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc
      841 gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta
      901 tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct
      961 acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc
     1021 tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa
     1081 caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa
     1141 aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa
     1201 aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt
     1261 ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaacc tgaggctatg
     1321 gcagggcctg ccgccccgac gttggctgcg agccctgggc cttcacccga acttgggggg
     1381 tggggtgggg aaaaggaaga aacgcgggcg tattggcccc aatggggtct cggtggggta
     1441 tcgacagagt gccagccctg ggaccgaacc ccgcgtttat gaacaaacga cccaacaccg
     1501 tgcgttttat tctgtctttt tattgccgtc atagcgcggg ttccttccgg tattgtctcc
     1561 ttccgtgttt cagttagcct ccccctaggg tgggcgaaga actccagcat gagatccccg
     1621 cgctggagga tcatccagcc ggcgtcccgg aaaacgattc cgaagcccaa cctttcatag
     1681 aaggcggcgg tggaatcgaa atctcgtgat ggcaggttgg gcgtcgcttg gtcggtcatt
     1741 tcgaacccca gagtcccgct cagaagaact cgtcaagaag gcgatagaag gcgatgcgct
     1801 gcgaatcggg agcggcgata ccgtaaagca cgaggaagcg gtcagcccat tcgccgccaa
     1861 gctcttcagc aatatcacgg gtagccaacg ctatgtcctg atagcggtcc gccacaccca
     1921 gccggccaca gtcgatgaat ccagaaaagc ggccattttc caccatgata ttcggcaagc
     1981 aggcatcgcc atgggtcacg acgagatcct cgccgtcggg catgctcgcc ttgagcctgg
     2041 cgaacagttc ggctggcgcg agcccctgat gctcttcgtc cagatcatcc tgatcgacaa
     2101 gaccggcttc catccgagta cgtgctcgct cgatgcgatg tttcgcttgg tggtcgaatg
     2161 ggcaggtagc cggatcaagc gtatgcagcc gccgcattgc atcagccatg atggatactt
     2221 tctcggcagg agcaaggtga gatgacagga gatcctgccc cggcacttcg cccaatagca
     2281 gccagtccct tcccgcttca gtgacaacgt cgagcacagc tgcgcaagga acgcccgtcg
     2341 tggccagcca cgatagccgc gctgcctcgt cttgcagttc attcagggca ccggacaggt
     2401 cggtcttgac aaaaagaacc gggcgcccct gcgctgacag ccggaacacg gcggcatcag
     2461 agcagccgat tgtctgttgt gcccagtcat agccgaatag cctctccacc caagcggccg
     2521 gagaacctgc gtgcaatcca tcttgttcaa tcatgcgaaa cgatcctcat cctgtctctt
     2581 gatcgatctt tgcaaaagcc taggcctcca aaaaagcctc ctcactactt ctggaatagc
     2641 tcagaggccg ggcctcggcc tctgcataaa taaaaaaaat tagtcagcca tggggcggag
     2701 aatgggcgga actgggcgga gttaggggcg ggatgggcgg agttaggggc gggactatgg
     2761 ttgctgacta attgagatgc atgctttgca tacttctgcc tgctggggag cctggggact
     2821 ttccacacct ggttgctgac taattgagat gcatgctttg catacttctg cctgctgggg
     2881 agcctgggga ctttccacac cctaactgac acacattcca cagctggttc tttccgcctc
     2941 aggactcttc ctttttcaat aaatcaatct aaagtatata tgagtaaact tggtctgaca
     3001 gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca
     3061 tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc
     3121 ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa
     3181 accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc
     3241 agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca
     3301 acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat
     3361 tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag
     3421 cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac
     3481 tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt
     3541 ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt
     3601 gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc
     3661 tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat
     3721 ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca
     3781 gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga
     3841 cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg
     3901 gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg
     3961 ttccgcgcac atttccccga aaagtgccac ctgacgcgcc ctgtagcggc gcattaagcg
     4021 cggcgggtgt ggtggttacg cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg
     4081 ctcctttcgc tttcttccct tcctttctcg ccacgttcgc cggctttccc cgtcaagctc
     4141 taaatcgggg gctcccttta gggttccgat ttagtgcttt acggcacctc gaccccaaaa
     4201 aacttgatta gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc
     4261 ctttgacgtt ggagtccacg ttctttaata gtggactctt gttccaaact ggaacaacac
     4321 tcaaccctat ctcggtctat tcttttgatt tataagggat tttgccgatt tcggcctatt
     4381 ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa ttttaacaaa atattaacgc
     4441 ttacaattta cgcgcgcgtt gacattgatt attgactagt tattaatagt aatcaattac
     4501 ggggtcatta gttcatagcc catatatgga gttccgcgtt acataactta cggtaaatgg
     4561 cccgcctggc tgaccgccca acgacccccg cccattgacg tcaataatga cgtatgttcc
     4621 catagtaacg ccaataggga ctttccattg acgtcaatgg gtggagtatt tacggtaaac
     4681 tgcccacttg gcagtacatc aagtgtatca tatgccaagt acgcccccta ttgacgtcaa
     4741 tgacggtaaa tggcccgcct ggcattatgc ccagtacatg accttatggg actttcctac
     4801 ttggcagtac atctacgtat tagtcatcgc tattaccatg gtgatgcggt tttggcagta
     4861 catcaatggg cgtggatagc ggtttgactc acggggattt ccaagtctcc accccattga
     4921 cgtcaatggg agtttgtttt ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa
     4981 ctccgcccca ttgacgcaaa tgggcggtag gcgtgtacgg tgggaggtct atataagcag
     5041 agctctctgg ctaactagag aacccactgc ttactggctt atcgaaatta atacgactca
     5101 ctatagggag acccaagctt ggtaccgagc tcggatccac tagtaacggc cgccagtgtg
     5161 ctggaattcg gcttggggat atccaccatg gagacagaca cactcctgct atgggtactg
     5221 ctgctctggg ttccaggttc cactggtgac tatccatatg atgttccaga ttatgctatg
     5281 gagatggcga gccagcaggg cgagacgatg tccggaccgc tgcacattgg tttgattccc
     5341 acagttggac cgtacctgct accgcatatt atccctatgc tgcaccagac ctttccaaag
     5401 ctggaaatgt atctgcatga agcacagacc caccagttac tggcgcaact ggacagcggc
     5461 aaactcgatt gcgtgatcct cgcactggtg aaagagagcg aagcattcat tgaagtgccg
     5521 ttgtttgatg agccaatgtt gctggctatc tatgaagatc acccgtgggc gaaccgcgaa
     5581 tgcgtaccga tggccgatct ggcaggggaa aaactgctga tgctggaaga tggtcactgt
     5641 ttgcgcgatc aggcaatggg tttctgtttt gaagccgggg cgctgagctc aacgggtgag
     5701 accttccagg ccttccgcac caccgacgtc ggccgcaagc tgatcatcga tcagaacgtt
     5761 tttatcgagg gtacgctgcc gatgggtgtc gtccgcccgc tgactgaagt cgagatggac
     5821 cattaccgcg agccgttcct gaatcctgtt gaccgcgagc cactgtggcg cttcccaaac
     5881 gagctgccaa tcgccggtga gccagcgaac atcgtcgcgc tggtcgaaga atacatggac
     5941 tggctgcacc agtcccctgt cccgaagctg ctgttctggg gcaccccagg cgttctgatc
     6001 ccaccggccg aagccgctcg cctggccaaa agcctgccta actgcaaggc tgtggacatc
     6061 ggcccgggtc tgaatctgct gcaagaagac aacccggacc tgatcggcag cgagatcgcg
     6121 cgctggctgt cgacgctcga gatttccggc ggaggaacag gtggttctgg tggaacaggg
     6181 ggtagcggag gtacaggagg aagtatggcg gagatcggaa ctggattccc gtttgatccg
     6241 cattatgtgg aagttctggg agagcgcatg cattatgtgg acgttggtcc tcgtgatggg
     6301 acaccagtgc tgttccttca cggcaatccg acatcgtcgt acgtgtggcg taatatcatc
     6361 ccgcacgttg cccccacgca ccgctgcatt gcccctgact taattggtat ggggaaaagt
     6421 gataagcctg atctggggta cttctttgac gaccacgtac gcttcatgga tgcttttatt
     6481 gaagcattgg gtttggagga agtagttttg gtgatccatg attggggtag tgctctgggg
     6541 ttccattggg ccaagcgtaa cccagaacgc gtgaaaggaa ttgcctttat ggagttcatc
     6601 cgtccgattc caacatggga cgaatggcga ccggaccaac tggatgaaga tacacacttc
     6661 cgcgcgacca gcctggaaac tctgcgcaac atggtggcgg caggtagcgg gatcacttta
     6721 ctgccagcgc tggctgtgcc gccggagcgc aaacgcgatg gggttgttta tctgccgtgc
     6781 attaagccgg aaccacgccg cactattggt ctggtttatc gtcctggctc accgctgcgc
     6841 agccgctatg agcagctggc agaggccatc cgcgcaagaa tggatggcca tttcgataaa
     6901 gttttaaaac aggcggtt
//
