LOCUS       sequence-414328-        4647 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of mCherry with Y70A mutation.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4647)
  AUTHORS   Sekhon H, Ha JH, Presti MF, Procopio SB, Jarvis AR, Mirsky PO, John 
            AM, Loh SN
  TITLE     Adaptable, turn-on maturation (ATOM) fluorescent biosensors for 
            multiplexed detection in cells.
  JOURNAL   Nat Methods. 2023 Nov 9. doi: 10.1038/s41592-023-02065-w.
  PUBMED    37945909
REFERENCE   2  (bases 1 to 4647)
  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: "Nat 
            Methods. 2023 Nov 9. doi: 10.1038/s41592-023-02065-w."
COMMENT     Sequence Label: pCMV-mCherry(Y70A)
FEATURES             Location/Qualifiers
     source          1..4647
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     63..82
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(381..400)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    425..472
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      801..1389
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1290..1309
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        1564..1867
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1868..2071
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2022..2042
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      2100..2109
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2106..2816
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMAGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(2251..2269)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     2519..2538
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     polyA_signal    2938..3059
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2975..2994)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3029..3048
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3066..3521)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3203..3224)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3415..3434
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3548..3652
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3654..4011
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3654..3845
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3677..3697)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3862..3997
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             join(4046..4647,1..193)
                     /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             join(4046..4647,1..193)
                     /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(4100..4119)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
ORIGIN
        1 ccgcttttct ggattcatcg actgtggccg gctgggtgtg gcggaccgct atcaggacat
       61 agcgttggct acccgtgata ttgctgaaga gcttggcggc gaatgggctg accgcttcct
      121 cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc gccttctatc gccttcttga
      181 cgagttcttc tgagcgggac tctggggttc gaaatgaccg accaagcgac gcccaacctg
      241 ccatcacgag atttcgattc caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt
      301 ttccgggacg ccggctggat gatcctccag cgcggggatc tcatgctgga gttcttcgcc
      361 caccctaggg ggaggctaac tgaaacacgg aaggagacaa taccggaagg aacccgcgct
      421 atgacggcaa taaaaagaca gaataaaacg cacggtgttg ggtcgtttgt tcataaacgc
      481 ggggttcggt cccagggctg gcactctgtc gataccccac cgagacccca ttggggccaa
      541 tacgcccgcg tttcttcctt ttccccaccc caccccccaa gttcgggtga aggcccaggg
      601 ctcgcagcca acgtcggggc ggcaggccct gccatagcct caggttactc atatatactt
      661 tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat
      721 aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta
      781 gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa
      841 acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt
      901 tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct tctagtgtag
      961 ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta
     1021 atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca
     1081 agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag
     1141 cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa
     1201 agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga
     1261 acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc
     1321 gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc
     1381 ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt
     1441 gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat taccgccatg
     1501 cattagttat taatagtaat caattacggg gtcattagtt catagcccat atatggagtt
     1561 ccgcgttaca taacttacgg taaatggccc gcctggctga ccgcccaacg acccccgccc
     1621 attgacgtca ataatgacgt atgttcccat agtaacgcca atagggactt tccattgacg
     1681 tcaatgggtg gagtatttac ggtaaactgc ccacttggca gtacatcaag tgtatcatat
     1741 gccaagtacg ccccctattg acgtcaatga cggtaaatgg cccgcctggc attatgccca
     1801 gtacatgacc ttatgggact ttcctacttg gcagtacatc tacgtattag tcatcgctat
     1861 taccatggtg atgcggtttt ggcagtacat caatgggcgt ggatagcggt ttgactcacg
     1921 gggatttcca agtctccacc ccattgacgt caatgggagt ttgttttggc accaaaatca
     1981 acgggacttt ccaaaatgtc gtaacaactc cgccccattg acgcaaatgg gcggtaggcg
     2041 tgtacggtgg gaggtctata taagcagagc tggtttagtg aaccgtcaga tccgctagcg
     2101 ccaccatggt gagcaagggc gaggaggata acatggccat catcaaggag ttcatgcgct
     2161 tcaaggtgca catggagggc tccgtgaacg gccacgagtt cgagatcgag ggcgagggcg
     2221 agggccgccc ctacgagggc acccagaccg ccaagctgaa ggtgaccaag ggtggccccc
     2281 tgcccttcgc ctgggacatc ctgtcccctc agttcatggc cggctccaag gcctacgtga
     2341 agcaccccgc cgacatcccc gactacttga agctgtcctt ccccgagggc ttcaagtggg
     2401 agcgcgtgat gaacttcgag gacggcggcg tggtgaccgt gacccaggac tcctccctgc
     2461 aggacggcga gttcatctac aaggtgaagc tgcgcggcac caacttcccc tccgacggcc
     2521 ccgtaatgca gaagaagacc atgggctggg aggcctcctc cgagcggatg taccccgagg
     2581 acggcgccct gaagggcgag atcaagcaga ggctgaagct gaaggacggc ggccactacg
     2641 acgctgaggt caagaccacc tacaaggcca agaagcccgt gcagctgccc ggcgcctaca
     2701 acgtcaacat caagttggac atcacctccc acaacgagga ctacaccatc gtggaacagt
     2761 acgaacgcgc cgagggccgc cactccaccg gcggcatgga cgagctgtac aagtgagcgg
     2821 ccgcgactct agatcataat cagccatacc acatttgtag aggttttact tgctttaaaa
     2881 aacctcccac acctccccct gaacctgaaa cataaaatga atgcaattgt tgttgttaac
     2941 ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat
     3001 aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttaa
     3061 ggcgtaaatt gtaagcgtta atattttgtt aaaattcgcg ttaaattttt gttaaatcag
     3121 ctcatttttt aaccaatagg ccgaaatcgg caaaatccct tataaatcaa aagaatagac
     3181 cgagataggg ttgagtgttg ttccagtttg gaacaagagt ccactattaa agaacgtgga
     3241 ctccaacgtc aaagggcgaa aaaccgtcta tcagggcgat ggcccactac gtgaaccatc
     3301 accctaatca agttttttgg ggtcgaggtg ccgtaaagca ctaaatcgga accctaaagg
     3361 gagcccccga tttagagctt gacggggaaa gccggcgaac gtggcgagaa aggaagggaa
     3421 gaaagcgaaa ggagcgggcg ctagggcgct ggcaagtgta gcggtcacgc tgcgcgtaac
     3481 caccacaccc gccgcgctta atgcgccgct acagggcgcg tcaggtggca cttttcgggg
     3541 aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct
     3601 catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga gtcctgaggc
     3661 ggaaagaacc agctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca
     3721 gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccaggtg tggaaagtcc
     3781 ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccata
     3841 gtcccgcccc taactccgcc catcccgccc ctaactccgc ccagttccgc ccattctccg
     3901 ccccatggct gactaatttt ttttatttat gcagaggccg aggccgcctc ggcctctgag
     3961 ctattccaga agtagtgagg aggctttttt ggaggcctag gcttttgcaa agatcgatca
     4021 agagacagga tgaggatcgt ttcgcatgat tgaacaagat ggattgcacg caggttctcc
     4081 ggccgcttgg gtggagaggc tattcggcta tgactgggca caacagacaa tcggctgctc
     4141 tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg gttctttttg tcaagaccga
     4201 cctgtccggt gccctgaatg aactgcaaga cgaggcagcg cggctatcgt ggctggccac
     4261 gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact gaagcgggaa gggactggct
     4321 gctattgggc gaagtgccgg ggcaggatct cctgtcatct caccttgctc ctgccgagaa
     4381 agtatccatc atggctgatg caatgcggcg gctgcatacg cttgatccgg ctacctgccc
     4441 attcgaccac caagcgaaac atcgcatcga gcgagcacgt actcggatgg aagccggtct
     4501 tgtcgatcag gatgatctgg acgaagagca tcaggggctc gcgccagccg aactgttcgc
     4561 caggctcaag gcgagcatgc ccgacggcga ggatctcgtc gtgacccatg gcgatgcctg
     4621 cttgccgaat atcatggtgg aaaatgg
//
