LOCUS       sequence-262557-        5596 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mitochondrial ratiometric pH probe with enhanced brightness fused to
            Smac/DIABLO fragment. Probe is localized in mitochondrial 
            intermembrane space. Mammalian cell expression..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5596)
  AUTHORS   Ermakova YG, Pak VV, Bogdanova YA, Kotlobay AA, Yampolsky IV, 
            Shokhina AG, Panova AS, Marygin RA, Staroverov DB, Bilan DS, Sies H,
            Belousov VV
  TITLE     SypHer3s: a genetically encoded fluorescent ratiometric probe with 
            enhanced brightness and an improved dynamic range.
  JOURNAL   Chem Commun (Camb). 2018 Mar 2. doi: 10.1039/c7cc08740c.
  PUBMED    29497715
REFERENCE   2  (bases 1 to 5596)
  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: "Chem Commun
            (Camb). 2018 Mar 2. doi: 10.1039/c7cc08740c."
COMMENT     Sequence Label: pC1-SypHer3s-IMS
FEATURES             Location/Qualifiers
     source          1..5596
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        61..364
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        365..568
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     519..539
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      597..606
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     regulatory      601..610
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     complement(1542..1563)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1803..1822)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     polyA_signal    2363..2484
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2400..2419)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2454..2473
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2491..2946)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2628..2649)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2840..2859
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2973..3077
                     /gene="bla"
                     /label=AmpR promoter
     enhancer        3079..3270
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3102..3122)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3308..3443
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3492..4286
                     /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             3492..4286
                     /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(3546..3565)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4156..4175
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4474..4493)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4518..4565
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4894..5482
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5383..5402
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg
       61 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
      121 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      181 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      241 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      301 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      361 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      421 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      481 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      541 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgcca
      601 ccatggcggc tctgaagagt tggctgtcgc gcagcgtaac ttcattcttc aggtacagac
      661 agtgtttgtg tgttcctgtt gtggctaact ttaagaagcg gtgtttctca gaattgataa
      721 gaccatggca caaaactgtg acgattggct ttggagtaac cctgtgtgcg gttcctatta
      781 agcttcgaat tctgcagtcg acggtaccgc gggcccggga tccacccgtc gccacctctg
      841 gaccgctgca cattggtttg attcccacag ttggaccgta cctgctaccg catattatcc
      901 ctatgctgca ccagaccttt ccaaagctgg aaatgtatct gcatgaagca cagacccacc
      961 agttactggc gcaactggac agcggcaaac tcgattgcgt gatcctcgcg ctggtgaaag
     1021 agagcgaagc attcattgaa gtgccgttgt ttgatgagcc aatgttgctg gctatctatg
     1081 aagatcaccc gtgggcgaac cgcgaatgcg taccgatggc cgatctggca ggggaaaaac
     1141 tgctgatgct ggaagatggt cactctttgc gcgatcaggc aatgtccgcc ggcttcaaca
     1201 gcgacaacgt ctatatcatg gccgacaagc agaagaacgg catcaaggcc aacttcaaga
     1261 tccgccacaa cgtcgaggac ggcagcgtgc agctcgccga ccactaccag cagaacaccc
     1321 ccatcggcga cggccccgtg ctgctgcccg acaaccacta cctgagcttc cagtccgtcc
     1381 tgagcaaaga ccccaacgag aagcgcgatc acatggtcct gcaggagttc gtgaccgccg
     1441 ccgggatcac tctcggcatg gacgagctgt acaacgtgga tggcggtagc ggtggcaccg
     1501 gcagcaaggg cgaggagctg ttcaccgggg tggtgcccat cctggtcgag ctggacggcg
     1561 acgtaaacgg ccacaagttc agcgtgtccg gcgagggcga gggcgatgcc acctacggca
     1621 agctgaccct gaagctgatc tgcaccaccg gcaagctgcc cgtgccctgg cccaccctcg
     1681 tgaccaccct cggctacggc ctgaagtgct tcgcccgcta ccccgaccac atgaagcagc
     1741 acgacttctt caagtccgcc atgcccgaag gctacgtcca ggagcgcacc atcttcttca
     1801 aggacgacgg caactacaag acccgcgccg aggtgaagtt cgagggcgac accctggtga
     1861 accgcatcga gctgaagggc atcggcttca aggaggacgg caacatcctg gggcacaagc
     1921 tggagtacaa cggcaccggt ttctgttttg aagccggggc ggatgaagat acacacttcc
     1981 gcgcgaccag cctggaaact ctgcgcaaca tggtggcggc aggtagcggg atcactttac
     2041 tgccagcgct ggctgtgccg ccggagcgca aacgcgatgg ggttgtttat ctgccgtgca
     2101 ttaagccgga accacgccgc actattggcc tggtttatcg tcctggctca ccgctgcgca
     2161 gccgctatga gcagctggca gaggccatcc gcgcaagaat ggatggacat ttcgataaag
     2221 ttttgaaaca ggcgttgtaa agcggccgcg actctagatc ataatcagcc ataccacatt
     2281 tgtagaggtt ttacttgctt taaaaaacct cccacacctc cccctgaacc tgaaacataa
     2341 aatgaatgca attgttgttg ttaacttgtt tattgcagct tataatggtt acaaataaag
     2401 caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt
     2461 gtccaaactc atcaatgtat cttaaggcgt aaattgtaag cgttaatatt ttgttaaaat
     2521 tcgcgttaaa tttttgttaa atcagctcat tttttaacca ataggccgaa atcggcaaaa
     2581 tcccttataa atcaaaagaa tagaccgaga tagggttgag tgttgttcca gtttggaaca
     2641 agagtccact attaaagaac gtggactcca acgtcaaagg gcgaaaaacc gtctatcagg
     2701 gcgatggccc actacgtgaa ccatcaccct aatcaagttt tttggggtcg aggtgccgta
     2761 aagcactaaa tcggaaccct aaagggagcc cccgatttag agcttgacgg ggaaagccgg
     2821 cgaacgtggc gagaaaggaa gggaagaaag cgaaaggagc gggcgctagg gcgctggcaa
     2881 gtgtagcggt cacgctgcgc gtaaccacca cacccgccgc gcttaatgcg ccgctacagg
     2941 gcgcgtcagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct
     3001 aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat
     3061 attgaaaaag gaagagtcct gaggcggaaa gaaccagctg tggaatgtgt gtcagttagg
     3121 gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta
     3181 gtcagcaacc aggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat
     3241 gcatctcaat tagtcagcaa ccatagtccc gcccctaact ccgcccatcc cgcccctaac
     3301 tccgcccatc ccgcccctaa ctccgcccag ttccgcccat tctccgcccc atggctgact
     3361 aatttttttt atttatgcag aggccgaggc cgcctcggcc tctgagctat tccagaagta
     3421 gtgaggaggc ttttttggag gcctaggctt ttgcaaagat cgatcaagag acaggatgag
     3481 gatcgtttcg catgattgaa caagatggat tgcacgcagg ttctccggcc gcttgggtgg
     3541 agaggctatt cggctatgac tgggcacaac agacaatcgg ctgctctgat gccgccgtgt
     3601 tccggctgtc agcgcagggg cgcccggttc tttttgtcaa gaccgacctg tccggtgccc
     3661 tgaatgaact gcaagacgag gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt
     3721 gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga ctggctgcta ttgggcgaag
     3781 tgccggggca ggatctcctg tcatctcacc ttgctcctgc cgagaaagta tccatcatgg
     3841 ctgatgcaat gcggcggctg catacgcttg atccggctac ctgcccattc gaccaccaag
     3901 cgaaacatcg catcgagcga gcacgtactc ggatggaagc cggtcttgtc gatcaggatg
     3961 atctggacga agagcatcag gggctcgcgc cagccgaact gttcgccagg ctcaaggcga
     4021 gcatgcccga cggcgaggat ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca
     4081 tggtggaaaa tggccgcttt tctggattca tcgactgtgg ccggctgggt gtggcggacc
     4141 gctatcagga catagcgttg gctacccgtg atattgctga agagcttggc ggcgaatggg
     4201 ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc atcgccttct
     4261 atcgccttct tgacgagttc ttctgagcgg gactctgggg ttcgaaatga ccgaccaagc
     4321 gacgcccaac ctgccatcac gagatttcga ttccaccgcc gccttctatg aaaggttggg
     4381 cttcggaatc gttttccggg acgccggctg gatgatcctc cagcgcgggg atctcatgct
     4441 ggagttcttc gcccacccta gggggaggct aactgaaaca cggaaggaga caataccgga
     4501 aggaacccgc gctatgacgg caataaaaag acagaataaa acgcacggtg ttgggtcgtt
     4561 tgttcataaa cgcggggttc ggtcccaggg ctggcactct gtcgataccc caccgagacc
     4621 ccattggggc caatacgccc gcgtttcttc cttttcccca ccccaccccc caagttcggg
     4681 tgaaggccca gggctcgcag ccaacgtcgg ggcggcaggc cctgccatag cctcaggtta
     4741 ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa
     4801 gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc
     4861 gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat
     4921 ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga
     4981 gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt
     5041 tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata
     5101 cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac
     5161 cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg
     5221 ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg
     5281 tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag
     5341 cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct
     5401 ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc
     5461 aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt
     5521 ttgctggcct tttgctcaca tgttctttcc tgcgttatcc cctgattctg tggataaccg
     5581 tattaccgcc atgcat
//
