LOCUS       sequence-409877-        6532 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression (cytosol) of the 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 6532)
  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 6532)
  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-mCherryEA
FEATURES             Location/Qualifiers
     source          1..6532
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             189..899
                     /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(334..352)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     602..621
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     polyA_signal    1025..1159
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1062..1081)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1116..1135
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(1182..1198)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1182..1198)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1195..1217)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1206..1222
                     /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(1230..1236,1237..1254,1255..1260))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     primer_bind     complement(1412..1429)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1583..2171)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1663..1682)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2342..3133,3134..3202))
                     /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     2965..2984
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      complement(3259..3772)
                     /direction=LEFT
                     /label=M13 ori
                     /note="M13 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3454..3475)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3666..3685
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3944..3962
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4000..4022)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4122..4141
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4373..4395
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     4387..4404
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     4388..4404
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     enhancer        4970..5349
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        5350..5553
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     5504..5524
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     5550..5574
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     intron          5689..6515
                     /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 ggtgagcaag ggcgaggagg ataacatggc catcatcaag gagttcatgc
      241 gcttcaaggt gcacatggag ggctccgtga acggccacga gttcgagatc gagggcgagg
      301 gcgagggccg cccctacgag ggcacccaga ccgccaagct gaaggtgacc aagggtggcc
      361 ccctgccctt cgcctgggac atcctgtccc ctcagttcat gtacggctcc aaggcctacg
      421 tgaagcaccc cgccgacatc cccgactact tgaagctgtc cttccccgag ggcttcaagt
      481 gggagcgcgt gatgaacttc gaggacggcg gcgtggtgac cgtgacccag gactcctccc
      541 tgcaggacgg cgagttcatc tacaaggtga agctgcgcgg caccaacttc ccctccgacg
      601 gccccgtaat gcagaagaag accatgggct gggaggcctc ctccgagcgg atgtaccccg
      661 aggacggcgc cctgaagggt gaggaaaagg cgaggctgaa gctgaaggac ggcggccact
      721 acgacgctga ggtcaagacc acctacaagg ccaagaagcc cgtgcagctg cccggcgcct
      781 acaacgtcaa catcaagttg gacatcacct cccacaacga ggactacacc atcgtggaac
      841 agtacgaacg cgccgagggt cgtcactcta ccggcggcat ggacgagctg tacaagtaat
      901 aaactcgacg aattcattga tcataatcag ccataccaca tttgtagagg ttttacttgc
      961 tttaaaaaac ctcccacacc tccccctgaa cctgaaacat aaaatgaatg caattgttgt
     1021 tgttaacttg tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt
     1081 cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt
     1141 atcttatcat gtctggatcc gtaccgagct cgcgtaatca tgtcatagct gtttcctgtg
     1201 tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa
     1261 gcctggggtg cctaatgagt gagctaacta cattaattgc gttgcgctca ctgcccgctt
     1321 tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag
     1381 gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg
     1441 ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat
     1501 caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta
     1561 aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa
     1621 atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc
     1681 cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt
     1741 ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca
     1801 gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg
     1861 accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat
     1921 cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta
     1981 cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct
     2041 gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac
     2101 aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa
     2161 aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa
     2221 actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt
     2281 taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca
     2341 gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca
     2401 tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc
     2461 ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa
     2521 accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc
     2581 agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca
     2641 acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat
     2701 tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag
     2761 cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac
     2821 tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt
     2881 ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt
     2941 gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc
     3001 tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat
     3061 ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca
     3121 gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga
     3181 cacggaaatg ttgaatactc atactcttcc tttttcaata agcggccgcg gccatgccgg
     3241 ccactagtct cgagggccac cccggttgat aatcagaaaa gccccaaaaa caggaagatt
     3301 gtataagcaa atatttaaat tgtaagcgtt aatattttgt taaaattcgc gttaaatttt
     3361 tgttaaatca gctcattttt taaccaatag gccgaaatcg gcaaaatccc ttataaatca
     3421 aaagaataga ccgagatagg gttgagtgtt gttccagttt ggaacaagag tccactatta
     3481 aagaacgtgg actccaacgt caaagggcga aaaaccgtct atcagggcga tggcccacta
     3541 cgtgaaccat caccctaatc aagttttttg gggtcgaggt gccgtaaagc actaaatcgg
     3601 aaccctaaag ggagcccccg atttagagct tgacggggaa agccggcgaa cgtggcgaga
     3661 aaggaaggga agaaagcgaa aggagcgggc gctagggcgc tggcaagtgt agcggtcacg
     3721 ctgcgcgtaa ccaccacacc cgccgcgctt aatgcgccgc tacagggcgc gtggggatcc
     3781 ctcgagttat tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga
     3841 atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc
     3901 tgacgtctaa gaaaccatta ttatcatgac attaacctat aaaaataggc gtatcacgag
     3961 gccctttcgt ctcgcgcgtt tcggtgatga cggtgaaaac ctctgacaca tgcagctccc
     4021 ggagacggtc acagcttgtc tgtaagcgga tgccgggagc agacaagccc gtcagggcgc
     4081 gtcagcgggt gttggcgggt gtcggggctg gcttaactat gcggcatcag agcagattgt
     4141 actgagagtg caccatatga agacgtcgcc tcctcactac ttctggaata gctcagaggc
     4201 cgaggcggcc tcggcctctg cataaataaa aaaaattagt cagggcgcca ttcgccattc
     4261 aggctgcgca actgttggga agggcgatcg gtgcgggcct cttcgctatt acgccagctg
     4321 gcgaaagggg gatgtgctgc aaggcgatta agttgggtaa cgccagggtt ttcccagtca
     4381 cgacgttgta aaacgacggc cagtgcccgt gtcaaggacg gtgactgcag tgaataataa
     4441 aatgtgtgtt tgtccgaaat acgcgttttg agatttctgt cgccgactaa attcatgtcg
     4501 cgcgatagtg gtgtttatcg ccgatagaga tggcgatatt ggaaaaatcg atatttgaaa
     4561 atatggcata ttgaaaatgt cgccgatgtg agtttctgtg taactgatat cgccattttt
     4621 ccaaaagtga tttttgggca tacgcgatat ctggcgatag cgcttatatc gtttacgggg
     4681 gatggcgata gacgactttg gtgacttggg cgattctgtg tgtcgcaaat atcgcagttt
     4741 cgatataggt gacagacgat atgaggctat atcgccgata gaggcgacat caagctggca
     4801 catggccaat gcatatcgat ctatacattg aatcaatatt ggccattagc catattattc
     4861 attggttata tagcataaat caatattggc tattggccat tgcatacgtt gtatccatat
     4921 cataatatgt acatttatat tggctcatgt ccaacattac cgccatgttg acattgatta
     4981 ttgactagtt attaatagta atcaattacg gggtcattag ttcatagccc atatatggag
     5041 ttccgcgtta cataacttac ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc
     5101 ccattgacgt caataatgac gtatgttccc atagtaacgc caatagggac tttccattga
     5161 cgtcaatggg tggagtattt acggtaaact gcccacttgg cagtacatca agtgtatcat
     5221 atgccaagta cgccccctat tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc
     5281 cagtacatga ccttatggga ctttcctact tggcagtaca tctacgtatt agtcatcgct
     5341 attaccatgg tgatgcggtt ttggcagtac atcaatgggc gtggatagcg gtttgactca
     5401 cggggatttc caagtctcca ccccattgac gtcaatggga gtttgttttg gcaccaaaat
     5461 caacgggact ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat gggcggtagg
     5521 cgtgtacggt gggaggtcta tataagcaga gctcgtttag tgaaccgtca gatcgcctgg
     5581 agacgccatc cacgctgttt tgacctccat agaagacacc gggaccgatc cagcctccgc
     5641 ggccgggaac ggtgcattgg aacgcggatt ccccgtgcca agagtgacgt aagtaccgcc
     5701 tatagagtct ataggcccac ccccttggct tcttatgcat gctatactgt ttttggcttg
     5761 gggtctatac acccccgctt cctcatgtta taggtgatgg tatagcttag cctataggtg
     5821 tgggttattg accattattg accactcccc tattggtgac gatactttcc attactaatc
     5881 cataacatgg ctctttgcca caactctctt tattggctat atgccaatac actgtccttc
     5941 agagactgac acggactctg tatttttaca ggatggggtc tcatttatta tttacaaatt
     6001 cacatataca acaccaccgt ccccagtgcc cgcagttttt attaaacata acgtgggatc
     6061 tccacgcgaa tctcgggtac gtgttccgga catgggctct tctccggtag cggcggagct
     6121 tctacatccg agccctgctc ccatgcctcc agcgactcat ggtcgctcgg cagctccttg
     6181 ctcctaacag tggaggccag acttaggcac agcacgatgc ccaccaccac cagtgtgccg
     6241 cacaaggccg tggcggtagg gtatgtgtct gaaaatgagc tcggggagcg ggcttgcacc
     6301 gctgacgcat ttggaagact taaggcagcg gcagaagaag atgcaggcag ctgagttgtt
     6361 gtgttctgat aagagtcaga ggtaactccc gttgcggtgc tgttaacggt ggagggcagt
     6421 gtagtctgag cagtactcgt tgctgccgcg cgcgccacca gacataatag ctgacagact
     6481 aacagactgt tcctttccat gggtcttttc tgcagtcacc gtccttgaca cg
//
