LOCUS       sequence-207649-        5386 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Ratiometric pH probe with enhanced brightness for mammalian cells 
            expression..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5386)
  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 5386)
  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
FEATURES             Location/Qualifiers
     source          1..5386
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    167..288
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(204..223)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     258..277
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(295..750)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(432..453)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     644..663
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        777..881
                     /gene="bla"
                     /label=AmpR promoter
     promoter        883..1240
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        883..1074
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(906..926)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      1091..1226
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             1275..2069
                     /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             1275..2069
                     /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(1329..1348)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     1939..1958
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(2257..2276)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    2301..2348
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      2677..3265
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3166..3185
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        3440..3743
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        3744..3947
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     3898..3918
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     complement(4691..4712)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(4952..4971)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
ORIGIN
        1 taatgagaat tctgcagtcg acggtaccgc gggcccggga tccaccggat ctagataact
       61 gatcataatc agccatacca catttgtaga ggttttactt gctttaaaaa acctcccaca
      121 cctccccctg aacctgaaac ataaaatgaa tgcaattgtt gttgttaact tgtttattgc
      181 agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt
      241 ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttaag gcgtaaattg
      301 taagcgttaa tattttgtta aaattcgcgt taaatttttg ttaaatcagc tcatttttta
      361 accaataggc cgaaatcggc aaaatccctt ataaatcaaa agaatagacc gagatagggt
      421 tgagtgttgt tccagtttgg aacaagagtc cactattaaa gaacgtggac tccaacgtca
      481 aagggcgaaa aaccgtctat cagggcgatg gcccactacg tgaaccatca ccctaatcaa
      541 gttttttggg gtcgaggtgc cgtaaagcac taaatcggaa ccctaaaggg agcccccgat
      601 ttagagcttg acggggaaag ccggcgaacg tggcgagaaa ggaagggaag aaagcgaaag
      661 gagcgggcgc tagggcgctg gcaagtgtag cggtcacgct gcgcgtaacc accacacccg
      721 ccgcgcttaa tgcgccgcta cagggcgcgt caggtggcac ttttcgggga aatgtgcgcg
      781 gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat
      841 aaccctgata aatgcttcaa taatattgaa aaaggaagag tcctgaggcg gaaagaacca
      901 gctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag caggcagaag
      961 tatgcaaagc atgcatctca attagtcagc aaccaggtgt ggaaagtccc caggctcccc
     1021 agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccatag tcccgcccct
     1081 aactccgccc atcccgcccc taactccgcc cagttccgcc cattctccgc cccatggctg
     1141 actaattttt tttatttatg cagaggccga ggccgcctcg gcctctgagc tattccagaa
     1201 gtagtgagga ggcttttttg gaggcctagg cttttgcaaa gatcgatcaa gagacaggat
     1261 gaggatcgtt tcgcatgatt gaacaagatg gattgcacgc aggttctccg gccgcttggg
     1321 tggagaggct attcggctat gactgggcac aacagacaat cggctgctct gatgccgccg
     1381 tgttccggct gtcagcgcag gggcgcccgg ttctttttgt caagaccgac ctgtccggtg
     1441 ccctgaatga actgcaagac gaggcagcgc ggctatcgtg gctggccacg acgggcgttc
     1501 cttgcgcagc tgtgctcgac gttgtcactg aagcgggaag ggactggctg ctattgggcg
     1561 aagtgccggg gcaggatctc ctgtcatctc accttgctcc tgccgagaaa gtatccatca
     1621 tggctgatgc aatgcggcgg ctgcatacgc ttgatccggc tacctgccca ttcgaccacc
     1681 aagcgaaaca tcgcatcgag cgagcacgta ctcggatgga agccggtctt gtcgatcagg
     1741 atgatctgga cgaagagcat caggggctcg cgccagccga actgttcgcc aggctcaagg
     1801 cgagcatgcc cgacggcgag gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata
     1861 tcatggtgga aaatggccgc ttttctggat tcatcgactg tggccggctg ggtgtggcgg
     1921 accgctatca ggacatagcg ttggctaccc gtgatattgc tgaagagctt ggcggcgaat
     1981 gggctgaccg cttcctcgtg ctttacggta tcgccgctcc cgattcgcag cgcatcgcct
     2041 tctatcgcct tcttgacgag ttcttctgag cgggactctg gggttcgaaa tgaccgacca
     2101 agcgacgccc aacctgccat cacgagattt cgattccacc gccgccttct atgaaaggtt
     2161 gggcttcgga atcgttttcc gggacgccgg ctggatgatc ctccagcgcg gggatctcat
     2221 gctggagttc ttcgcccacc ctagggggag gctaactgaa acacggaagg agacaatacc
     2281 ggaaggaacc cgcgctatga cggcaataaa aagacagaat aaaacgcacg gtgttgggtc
     2341 gtttgttcat aaacgcgggg ttcggtccca gggctggcac tctgtcgata ccccaccgag
     2401 accccattgg ggccaatacg cccgcgtttc ttccttttcc ccaccccacc ccccaagttc
     2461 gggtgaaggc ccagggctcg cagccaacgt cggggcggca ggccctgcca tagcctcagg
     2521 ttactcatat atactttaga ttgatttaaa acttcatttt taatttaaaa ggatctaggt
     2581 gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg
     2641 agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt
     2701 aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca
     2761 agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac
     2821 tgttcttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac
     2881 atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct
     2941 taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg
     3001 gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga gatacctaca
     3061 gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt
     3121 aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta
     3181 tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc
     3241 gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc
     3301 cttttgctgg ccttttgctc acatgttctt tcctgcgtta tcccctgatt ctgtggataa
     3361 ccgtattacc gccatgcatt agttattaat agtaatcaat tacggggtca ttagttcata
     3421 gcccatatat ggagttccgc gttacataac ttacggtaaa tggcccgcct ggctgaccgc
     3481 ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt tcccatagta acgccaatag
     3541 ggactttcca ttgacgtcaa tgggtggagt atttacggta aactgcccac ttggcagtac
     3601 atcaagtgta tcatatgcca agtacgcccc ctattgacgt caatgacggt aaatggcccg
     3661 cctggcatta tgcccagtac atgaccttat gggactttcc tacttggcag tacatctacg
     3721 tattagtcat cgctattacc atggtgatgc ggttttggca gtacatcaat gggcgtggat
     3781 agcggtttga ctcacgggga tttccaagtc tccaccccat tgacgtcaat gggagtttgt
     3841 tttggcacca aaatcaacgg gactttccaa aatgtcgtaa caactccgcc ccattgacgc
     3901 aaatgggcgg taggcgtgta cggtgggagg tctatataag cagagctggt ttagtgaacc
     3961 gtcagatccg ctagccgcca ccatgtccgg accgctgcac attggtttga ttcccacagt
     4021 tggaccgtac ctgctaccgc atattatccc tatgctgcac cagacctttc caaagctgga
     4081 aatgtatctg catgaagcac agacccacca gttactggcg caactggaca gcggcaaact
     4141 cgattgcgtg atcctcgcgc tggtgaaaga gagcgaagca ttcattgaag tgccgttgtt
     4201 tgatgagcca atgttgctgg ctatctatga agatcacccg tgggcgaacc gcgaatgcgt
     4261 accgatggcc gatctggcag gggaaaaact gctgatgctg gaagatggtc actctttgcg
     4321 cgatcaggca atgtccgccg gcttcaacag cgacaacgtc tatatcatgg ccgacaagca
     4381 gaagaacggc atcaaggcca acttcaagat ccgccacaac gtcgaggacg gcagcgtgca
     4441 gctcgccgac cactaccagc agaacacccc catcggcgac ggccccgtgc tgctgcccga
     4501 caaccactac ctgagcttcc agtccgtcct gagcaaagac cccaacgaga agcgcgatca
     4561 catggtcctg caggagttcg tgaccgccgc cgggatcact ctcggcatgg acgagctgta
     4621 caacgtggat ggcggtagcg gtggcaccgg cagcaagggc gaggagctgt tcaccggggt
     4681 ggtgcccatc ctggtcgagc tggacggcga cgtaaacggc cacaagttca gcgtgtccgg
     4741 cgagggcgag ggcgatgcca cctacggcaa gctgaccctg aagctgatct gcaccaccgg
     4801 caagctgccc gtgccctggc ccaccctcgt gaccaccctc ggctacggcc tgaagtgctt
     4861 cgcccgctac cccgaccaca tgaagcagca cgacttcttc aagtccgcca tgcccgaagg
     4921 ctacgtccag gagcgcacca tcttcttcaa ggacgacggc aactacaaga cccgcgccga
     4981 ggtgaagttc gagggcgaca ccctggtgaa ccgcatcgag ctgaagggca tcggcttcaa
     5041 ggaggacggc aacatcctgg ggcacaagct ggagtacaac ggcaccggtt tctgttttga
     5101 agccggggcg gatgaagata cacacttccg cgcgaccagc ctggaaactc tgcgcaacat
     5161 ggtggcggca ggtagcggga tcactttact gccagcgctg gctgtgccgc cggagcgcaa
     5221 acgcgatggg gttgtttatc tgccgtgcat taagccggaa ccacgccgca ctattggcct
     5281 ggtttatcgt cctggctcac cgctgcgcag ccgctatgag cagctggcag aggccatccg
     5341 cgcaagaatg gatggccatt tcgataaagt tttaaaacag gcggtt
//
