LOCUS       sequence-203922-        5388 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 5388)
  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 5388)
  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..5388
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    168..289
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(205..224)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     259..278
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(296..751)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(433..454)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     645..664
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        778..882
                     /gene="bla"
                     /label=AmpR promoter
     promoter        884..1241
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        884..1075
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(907..927)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      1092..1227
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             1276..2070
                     /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             1276..2070
                     /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(1330..1349)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     1940..1959
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(2258..2277)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    2302..2349
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      2678..3266
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3167..3186
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        3441..3744
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        3745..3948
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     3899..3919
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     complement(4690..4711)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(4951..4970)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
ORIGIN
        1 aagcttcgaa ttctgcagtc gacggtaccg cgggcccggg atccaccgga tctagataac
       61 tgatcataat cagccatacc acatttgtag aggttttact tgctttaaaa aacctcccac
      121 acctccccct gaacctgaaa cataaaatga atgcaattgt tgttgttaac ttgtttattg
      181 cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt
      241 tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttaa cgcgtaaatt
      301 gtaagcgtta atattttgtt aaaattcgcg ttaaattttt gttaaatcag ctcatttttt
      361 aaccaatagg ccgaaatcgg caaaatccct tataaatcaa aagaatagac cgagataggg
      421 ttgagtgttg ttccagtttg gaacaagagt ccactattaa agaacgtgga ctccaacgtc
      481 aaagggcgaa aaaccgtcta tcagggcgat ggcccactac gtgaaccatc accctaatca
      541 agttttttgg ggtcgaggtg ccgtaaagca ctaaatcgga accctaaagg gagcccccga
      601 tttagagctt gacggggaaa gccggcgaac gtggcgagaa aggaagggaa gaaagcgaaa
      661 ggagcgggcg ctagggcgct ggcaagtgta gcggtcacgc tgcgcgtaac caccacaccc
      721 gccgcgctta atgcgccgct acagggcgcg tcaggtggca cttttcgggg aaatgtgcgc
      781 ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa
      841 taaccctgat aaatgcttca ataatattga aaaaggaaga gtcctgaggc ggaaagaacc
      901 agctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa
      961 gtatgcaaag catgcatctc aattagtcag caaccaggtg tggaaagtcc ccaggctccc
     1021 cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccata gtcccgcccc
     1081 taactccgcc catcccgccc ctaactccgc ccagttccgc ccattctccg ccccatggct
     1141 gactaatttt ttttatttat gcagaggccg aggccgcctc ggcctctgag ctattccaga
     1201 agtagtgagg aggctttttt ggaggcctag gcttttgcaa agatcgatca agagacagga
     1261 tgaggatcgt ttcgcatgat tgaacaagat ggattgcacg caggttctcc ggccgcttgg
     1321 gtggagaggc tattcggcta tgactgggca caacagacaa tcggctgctc tgatgccgcc
     1381 gtgttccggc tgtcagcgca ggggcgcccg gttctttttg tcaagaccga cctgtccggt
     1441 gccctgaatg aactgcaaga cgaggcagcg cggctatcgt ggctggccac gacgggcgtt
     1501 ccttgcgcag ctgtgctcga cgttgtcact gaagcgggaa gggactggct gctattgggc
     1561 gaagtgccgg ggcaggatct cctgtcatct caccttgctc ctgccgagaa agtatccatc
     1621 atggctgatg caatgcggcg gctgcatacg cttgatccgg ctacctgccc attcgaccac
     1681 caagcgaaac atcgcatcga gcgagcacgt actcggatgg aagccggtct tgtcgatcag
     1741 gatgatctgg acgaagagca tcaggggctc gcgccagccg aactgttcgc caggctcaag
     1801 gcgagcatgc ccgacggcga ggatctcgtc gtgacccatg gcgatgcctg cttgccgaat
     1861 atcatggtgg aaaatggccg cttttctgga ttcatcgact gtggccggct gggtgtggcg
     1921 gaccgctatc aggacatagc gttggctacc cgtgatattg ctgaagagct tggcggcgaa
     1981 tgggctgacc gcttcctcgt gctttacggt atcgccgctc ccgattcgca gcgcatcgcc
     2041 ttctatcgcc ttcttgacga gttcttctga gcgggactct ggggttcgaa atgaccgacc
     2101 aagcgacgcc caacctgcca tcacgagatt tcgattccac cgccgccttc tatgaaaggt
     2161 tgggcttcgg aatcgttttc cgggacgccg gctggatgat cctccagcgc ggggatctca
     2221 tgctggagtt cttcgcccac cctaggggga ggctaactga aacacggaag gagacaatac
     2281 cggaaggaac ccgcgctatg acggcaataa aaagacagaa taaaacgcac ggtgttgggt
     2341 cgtttgttca taaacgcggg gttcggtccc agggctggca ctctgtcgat accccaccga
     2401 gaccccattg gggccaatac gcccgcgttt cttccttttc cccaccccac cccccaagtt
     2461 cgggtgaagg cccagggctc gcagccaacg tcggggcggc aggccctgcc atagcctcag
     2521 gttactcata tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg
     2581 tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact
     2641 gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg
     2701 taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc
     2761 aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata
     2821 ctgtccttct agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta
     2881 catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc
     2941 ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg
     3001 ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac
     3061 agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg
     3121 taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt
     3181 atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct
     3241 cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg
     3301 ccttttgctg gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtggata
     3361 accgtattac cgccatgcat tagttattaa tagtaatcaa ttacggggtc attagttcat
     3421 agcccatata tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg
     3481 cccaacgacc cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata
     3541 gggactttcc attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta
     3601 catcaagtgt atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc
     3661 gcctggcatt atgcccagta catgacctta tgggactttc ctacttggca gtacatctac
     3721 gtattagtca tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga
     3781 tagcggtttg actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg
     3841 ttttggcacc aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg
     3901 caaatgggcg gtaggcgtgt acggtgggag gtctatataa gcagagctgg tttagtgaac
     3961 cgtcagatcc gctagcgcca catgtccgga ccgctgcaca ttggtttgat tcccacagtt
     4021 ggaccgtacc tgctaccgca tattatccct atgctgcacc agacctttcc aaagctggaa
     4081 atgtatctgc atgaagcaca gacccaccag ttactggcgc aactggacag cggcaaactc
     4141 gattgcgtga tcctcgcgct ggtgaaagag agcgaagcat tcattgaagt gccgttgttt
     4201 gatgagccaa tgttgctggc tatctatgaa gatcacccgt gggcgaaccg cgaatgcgta
     4261 ccgatggccg atctggcagg ggaaaaactg ctgatgctgg aagatggtca ctctttgcgc
     4321 gatcaggcaa tgtccgccgg cttcaacagc gacaacgtct atatcatggc cgacaagcag
     4381 aagaacggca tcaaggccaa cttcaagatc cgccacaacg tcgaggacgg cagcgtgcag
     4441 ctcgccgacc actaccagca gaacaccccc atcggcgacg gccccgtgct gctgcccgac
     4501 aaccactacc tgagcttcca gtccgtcctg agcaaagacc ccaacgagaa gcgcgatcac
     4561 atggtcctgc aggagttcgt gaccgccgcc gggatcactc tcggcatgga cgagctgtac
     4621 aacgtggatg gcggtagcgg tggcaccggc agcaagggcg aggagctgtt caccggggtg
     4681 gtgcccatcc tggtcgagct ggacggcgac gtaaacggcc acaagttcag cgtgtccggc
     4741 gagggcgagg gcgatgccac ctacggcaag ctgaccctga agctgatctg caccaccggc
     4801 aagctgcccg tgccctggcc caccctcgtg accaccctcg gctacggcct gaagtgcttc
     4861 gcccgctacc ccgaccacat gaagcagcac gacttcttca agtccgccat gcccgaaggc
     4921 tacgtccagg agcgcaccat cttcttcaag gacgacggca actacaagac ccgcgccgag
     4981 gtgaagttcg agggcgacac cctggtgaac cgcatcgagc tgaagggcat cggcttcaag
     5041 gaggacggca acatcctggg gcacaagctg gagtacaacg gcaccggttt ctgttttgaa
     5101 gccggggcgg atgaagatac acacttccgc gcgaccagcc tggaaactct gcgcaacatg
     5161 gtggcggcag gtagcgggat cactttactg ccagcgctgg ctgtgccgcc ggagcgcaaa
     5221 cgcgatgggg ttgtttatct gccgtgcatt aagccggaac cacgccgcac tattggcctg
     5281 gtttatcgtc ctggctcacc gctgcgcagc cgctatgagc agctggcaga ggccatccgc
     5341 gcaagaatgg atggccattt cgataaagtt ttaaaacagg cggtttaa
//
