LOCUS       sequence-409878-        6946 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mitochondria-targeted mCherry(I158E/Q160A) mutant as a red 
            fluorescent pH sensor.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6946)
  AUTHORS   Rajendran M, Claywell B, Haynes EP, Scales U, Henning CK, Tantama M
  TITLE     Imaging pH Dynamics Simultaneously in Two Cellular Compartments 
            Using a Ratiometric pH-Sensitive Mutant of mCherry.
  JOURNAL   ACS Omega. 2018 Aug 31;3(8):9476-9486. doi: 
            10.1021/acsomega.8b00655. Epub 2018 Aug 20.
  PUBMED    30197999
REFERENCE   2  (bases 1 to 6946)
  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"; doi: 
            "10.1021/acsomega.8b00655"; journalName: "ACS Omega"; date: 
            "2018-08-31- 31"; volume: "3"; issue: "8"; pages: "9476-9486"
COMMENT     Sequence Label: GW1-mito-mCherryEA
FEATURES             Location/Qualifiers
     source          1..6946
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             189..263
                     /codon_start=1
                     /product="mitochondrial presequence of human cytochrome c 
                     oxidase subunit VIII (Rizutto et al., 1989)"
                     /label=COX8 presequence
                     /translation="MSVLTPLLLRGLTGSARRLPVPRAK"
     CDS             285..359
                     /codon_start=1
                     /product="mitochondrial presequence of human cytochrome c 
                     oxidase subunit VIII (Rizutto et al., 1989)"
                     /label=COX8 presequence
                     /translation="MSVLTPLLLRGLTGSARRLPVPRAK"
     CDS             381..455
                     /codon_start=1
                     /product="mitochondrial presequence of human cytochrome c 
                     oxidase subunit VIII (Rizutto et al., 1989)"
                     /label=COX8 presequence
                     /translation="MSVLTPLLLRGLTGSARRLPVPRAK"
     CDS             477..551
                     /codon_start=1
                     /product="mitochondrial presequence of human cytochrome c 
                     oxidase subunit VIII (Rizutto et al., 1989)"
                     /label=COX8 presequence
                     /translation="MSVLTPLLLRGLTGSARRLPVPRAK"
     CDS             606..1316
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEEKARLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(751..769)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     1019..1038
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     polyA_signal    1439..1573
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1476..1495)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1530..1549
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(1596..1612)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1596..1612)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1609..1631)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1620..1636
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(1644..1650,1651..1668,1669..1674))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     primer_bind     complement(1826..1843)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1997..2585)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2077..2096)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2756..3547,3548..3616))
                     /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     3379..3398
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      complement(3673..4186)
                     /direction=LEFT
                     /label=M13 ori
                     /note="M13 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3868..3889)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4080..4099
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4358..4376
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4414..4436)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4536..4555
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4787..4809
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     4801..4818
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     4802..4818
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     enhancer        5384..5763
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        5764..5967
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     5918..5938
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     5964..5988
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     intron          6103..6929
                     /label=CMV intron A
                     /note="human cytomegalovirus intron A (Chapman et al., 
                     1991)"
ORIGIN
        1 aagcttcccg gccacagcaa tcgatccatg acacaaccag tgacaagcag tcccccagtg
       61 aaaccgcatc cgcatccgag tccgataccg ataaagattc tgaatcggag tgagtgccgc
      121 gtccgagagc gttccctgtc cacgtccacc atcggcggag caggtgtgcc tgaggcctct
      181 agaccaccat gagcgtgctg acaccattgt tgctcagagg cctcactggt tcagccaggc
      241 ggctgccagt cccaagagct aagatccata gtctgggaga tccaatgtca gttctcactc
      301 cccttttgct taggggactg accggaagcg ctcggcggtt gcctgtccca cgcgccaaga
      361 ttcactccct gggcgaccct atgagtgtcc ttactcctct tttgctgaga ggcttgacag
      421 ggagcgctcg gaggctccca gtgcctcgcg caaagatcca ttctctcgga gatcccatgt
      481 ccgtcctcac tcctctgctg ctgcgcgggc ttacaggttc cgctcgccgc ctgcctgttc
      541 ctcgggcaaa gatacacagt ctgggcgatc ctcaccggcg tggaggagca tctggtggag
      601 ctagtatggt gagcaagggc gaggaggata acatggccat catcaaggag ttcatgcgct
      661 tcaaggtgca catggagggc tccgtgaacg gccacgagtt cgagatcgag ggcgagggcg
      721 agggccgccc ctacgagggc acccagaccg ccaagctgaa ggtgaccaag ggtggccccc
      781 tgcccttcgc ctgggacatc ctgtcccctc agttcatgta cggctccaag gcctacgtga
      841 agcaccccgc cgacatcccc gactacttga agctgtcctt ccccgagggc ttcaagtggg
      901 agcgcgtgat gaacttcgag gacggcggcg tggtgaccgt gacccaggac tcctccctgc
      961 aggacggcga gttcatctac aaggtgaagc tgcgcggcac caacttcccc tccgacggcc
     1021 ccgtaatgca gaagaagacc atgggctggg aggcctcctc cgagcggatg taccccgagg
     1081 acggcgccct gaagggtgag gaaaaggcga ggctgaagct gaaggacggc ggccactacg
     1141 acgctgaggt caagaccacc tacaaggcca agaagcccgt gcagctgccc ggcgcctaca
     1201 acgtcaacat caagttggac atcacctccc acaacgagga ctacaccatc gtggaacagt
     1261 acgaacgcgc cgagggtcgt cactctaccg gcggcatgga cgagctgtac aagtaaacac
     1321 gacgaattca ttgatcataa tcagccatac cacatttgta gaggttttac ttgctttaaa
     1381 aaacctccca cacctccccc tgaacctgaa acataaaatg aatgcaattg ttgttgttaa
     1441 cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa
     1501 taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta
     1561 tcatgtctgg atccgtaccg agctcgcgta atcatgtcat agctgtttcc tgtgtgaaat
     1621 tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg taaagcctgg
     1681 ggtgcctaat gagtgagcta actacattaa ttgcgttgcg ctcactgccc gctttccagt
     1741 cgggaaacct gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt
     1801 tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc
     1861 tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg
     1921 ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg
     1981 ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac
     2041 gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg
     2101 gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct
     2161 ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg
     2221 tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct
     2281 gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac
     2341 tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt
     2401 tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc
     2461 tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca
     2521 ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat
     2581 ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac
     2641 gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt
     2701 aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc
     2761 aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg
     2821 cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg
     2881 ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc
     2941 cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta
     3001 ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg
     3061 ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct
     3121 ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta
     3181 gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg
     3241 ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga
     3301 ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt
     3361 gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca
     3421 ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt
     3481 cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt
     3541 ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga
     3601 aatgttgaat actcatactc ttcctttttc aataagcggc cgcggccatg ccggccacta
     3661 gtctcgaggg ccaccccggt tgataatcag aaaagcccca aaaacaggaa gattgtataa
     3721 gcaaatattt aaattgtaag cgttaatatt ttgttaaaat tcgcgttaaa tttttgttaa
     3781 atcagctcat tttttaacca ataggccgaa atcggcaaaa tcccttataa atcaaaagaa
     3841 tagaccgaga tagggttgag tgttgttcca gtttggaaca agagtccact attaaagaac
     3901 gtggactcca acgtcaaagg gcgaaaaacc gtctatcagg gcgatggccc actacgtgaa
     3961 ccatcaccct aatcaagttt tttggggtcg aggtgccgta aagcactaaa tcggaaccct
     4021 aaagggagcc cccgatttag agcttgacgg ggaaagccgg cgaacgtggc gagaaaggaa
     4081 gggaagaaag cgaaaggagc gggcgctagg gcgctggcaa gtgtagcggt cacgctgcgc
     4141 gtaaccacca cacccgccgc gcttaatgcg ccgctacagg gcgcgtgggg atccctcgag
     4201 ttattattga agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat
     4261 ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgacgt
     4321 ctaagaaacc attattatca tgacattaac ctataaaaat aggcgtatca cgaggccctt
     4381 tcgtctcgcg cgtttcggtg atgacggtga aaacctctga cacatgcagc tcccggagac
     4441 ggtcacagct tgtctgtaag cggatgccgg gagcagacaa gcccgtcagg gcgcgtcagc
     4501 gggtgttggc gggtgtcggg gctggcttaa ctatgcggca tcagagcaga ttgtactgag
     4561 agtgcaccat atgaagacgt cgcctcctca ctacttctgg aatagctcag aggccgaggc
     4621 ggcctcggcc tctgcataaa taaaaaaaat tagtcagggc gccattcgcc attcaggctg
     4681 cgcaactgtt gggaagggcg atcggtgcgg gcctcttcgc tattacgcca gctggcgaaa
     4741 gggggatgtg ctgcaaggcg attaagttgg gtaacgccag ggttttccca gtcacgacgt
     4801 tgtaaaacga cggccagtgc ccgtgtcaag gacggtgact gcagtgaata ataaaatgtg
     4861 tgtttgtccg aaatacgcgt tttgagattt ctgtcgccga ctaaattcat gtcgcgcgat
     4921 agtggtgttt atcgccgata gagatggcga tattggaaaa atcgatattt gaaaatatgg
     4981 catattgaaa atgtcgccga tgtgagtttc tgtgtaactg atatcgccat ttttccaaaa
     5041 gtgatttttg ggcatacgcg atatctggcg atagcgctta tatcgtttac gggggatggc
     5101 gatagacgac tttggtgact tgggcgattc tgtgtgtcgc aaatatcgca gtttcgatat
     5161 aggtgacaga cgatatgagg ctatatcgcc gatagaggcg acatcaagct ggcacatggc
     5221 caatgcatat cgatctatac attgaatcaa tattggccat tagccatatt attcattggt
     5281 tatatagcat aaatcaatat tggctattgg ccattgcata cgttgtatcc atatcataat
     5341 atgtacattt atattggctc atgtccaaca ttaccgccat gttgacattg attattgact
     5401 agttattaat agtaatcaat tacggggtca ttagttcata gcccatatat ggagttccgc
     5461 gttacataac ttacggtaaa tggcccgcct ggctgaccgc ccaacgaccc ccgcccattg
     5521 acgtcaataa tgacgtatgt tcccatagta acgccaatag ggactttcca ttgacgtcaa
     5581 tgggtggagt atttacggta aactgcccac ttggcagtac atcaagtgta tcatatgcca
     5641 agtacgcccc ctattgacgt caatgacggt aaatggcccg cctggcatta tgcccagtac
     5701 atgaccttat gggactttcc tacttggcag tacatctacg tattagtcat cgctattacc
     5761 atggtgatgc ggttttggca gtacatcaat gggcgtggat agcggtttga ctcacgggga
     5821 tttccaagtc tccaccccat tgacgtcaat gggagtttgt tttggcacca aaatcaacgg
     5881 gactttccaa aatgtcgtaa caactccgcc ccattgacgc aaatgggcgg taggcgtgta
     5941 cggtgggagg tctatataag cagagctcgt ttagtgaacc gtcagatcgc ctggagacgc
     6001 catccacgct gttttgacct ccatagaaga caccgggacc gatccagcct ccgcggccgg
     6061 gaacggtgca ttggaacgcg gattccccgt gccaagagtg acgtaagtac cgcctataga
     6121 gtctataggc ccaccccctt ggcttcttat gcatgctata ctgtttttgg cttggggtct
     6181 atacaccccc gcttcctcat gttataggtg atggtatagc ttagcctata ggtgtgggtt
     6241 attgaccatt attgaccact cccctattgg tgacgatact ttccattact aatccataac
     6301 atggctcttt gccacaactc tctttattgg ctatatgcca atacactgtc cttcagagac
     6361 tgacacggac tctgtatttt tacaggatgg ggtctcattt attatttaca aattcacata
     6421 tacaacacca ccgtccccag tgcccgcagt ttttattaaa cataacgtgg gatctccacg
     6481 cgaatctcgg gtacgtgttc cggacatggg ctcttctccg gtagcggcgg agcttctaca
     6541 tccgagccct gctcccatgc ctccagcgac tcatggtcgc tcggcagctc cttgctccta
     6601 acagtggagg ccagacttag gcacagcacg atgcccacca ccaccagtgt gccgcacaag
     6661 gccgtggcgg tagggtatgt gtctgaaaat gagctcgggg agcgggcttg caccgctgac
     6721 gcatttggaa gacttaaggc agcggcagaa gaagatgcag gcagctgagt tgttgtgttc
     6781 tgataagagt cagaggtaac tcccgttgcg gtgctgttaa cggtggaggg cagtgtagtc
     6841 tgagcagtac tcgttgctgc cgcgcgcgcc accagacata atagctgaca gactaacaga
     6901 ctgttccttt ccatgggtct tttctgcagt caccgtcctt gacacg
//
