LOCUS       sequence-478299-        5938 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses mCherry tagged methyl-CpG recognition motif targeted to 
            nucleus in mammalian cells..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5938)
  AUTHORS   Zhao H, Ma D, Xie J, Sanchez O, Huang F, Yuan C
  TITLE     Live-Cell Probe for In Situ Single-Cell Monitoring of Mitochondrial 
            DNA Methylation.
  JOURNAL   ACS Sens. 2021 Oct 22;6(10):3575-3586. doi: 
            10.1021/acssensors.1c00731. Epub 2021 Sep 29.
  PUBMED    34586782
REFERENCE   2  (bases 1 to 5938)
  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/acssensors.1c00731"; journalName: "ACS Sens"; date: 
            "2021-10-22- 22"; volume: "6"; issue: "10"; pages: "3575-3586"
COMMENT     Sequence Label: Nuc-meCpG
FEATURES             Location/Qualifiers
     source          1..5938
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        376..579
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     530..550
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     576..600
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             678..698
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             968..988
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     regulatory      1475..1484
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1481..2191
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(1626..1644)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     1894..1913
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     regulatory      2220..2229
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     polyA_signal    2233..2367
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2270..2289)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2324..2343
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     promoter        2436..2793
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2436..2627
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2459..2479)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      2644..2779
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(2813..2830)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2813..2829)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2822..2844)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      3042..3497
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3129..3148)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3339..3360
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3514..3533)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3633..3655
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3693..3711)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3779..3883
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3884..3952,3953..4744)
                     /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     complement(4102..4121)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4915..5503
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5404..5423
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5657..5674
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    5791..5812
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(5827..5832,5833..5850,5851..5857)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5865..5881
                     /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)."
     primer_bind     5870..5892
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     5889..5905
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5889..5905
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     enhancer        join(5934..5938,1..375)
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 tgattattga ctagttatta atagtaatca attacggggt cattagttca tagcccatat
       61 atggagttcc gcgttacata acttacggta aatggcccgc ctggctgacc gcccaacgac
      121 ccccgcccat tgacgtcaat aatgacgtat gttcccatag taacgccaat agggactttc
      181 cattgacgtc aatgggtgga gtatttacgg taaactgccc acttggcagt acatcaagtg
      241 tatcatatgc caagtacgcc ccctattgac gtcaatgacg gtaaatggcc cgcctggcat
      301 tatgcccagt acatgacctt atgggacttt cctacttggc agtacatcta cgtattagtc
      361 atcgctatta ccatggtgat gcggttttgg cagtacatca atgggcgtgg atagcggttt
      421 gactcacggg gatttccaag tctccacccc attgacgtca atgggagttt gttttggcac
      481 caaaatcaac gggactttcc aaaatgtcgt aacaactccg ccccattgac gcaaatgggc
      541 ggtaggcgtg tacggtggga ggtctatata agcagagctc gtttagtgaa ccgtcagatc
      601 gcctggagac gccatccacg ctgttttgac ctccatagaa gacaccggga ccgatccagc
      661 ctccgcggcc cgggatgcca aaaaagaaga gaaaggtagg tagcggcagc ggtagcggtt
      721 ccggaggcag cgaattcgct gaggactggc tggacagccc ggccctgggc cctggctgga
      781 agcgccgcga agtctttcgc aagtcagggg ccaccgcggg acgctcagac acctattacc
      841 agagccccac aggagacagg atccgaagca aagttgagct gactcgatac ctgggccctg
      901 cgggcgatct caccctcttc gacttcaaac aaggcatctt gggcggtggc caattcgtac
      961 cgggatgcca aaaaagaaga gaaaggtagg tagcggcagc ggtagcggtt ccggaggcag
     1021 cgaattcgct gaggactggc tggacagccc ggccctgggc cctggctgga agcgccgcga
     1081 agtctttcgc aagtcagggg ccaccgcggg acgctcagac acctattacc agagccccac
     1141 aggagacagg atccgaagca aagttgagct gactcgatac ctgggccctg cgggcgatct
     1201 caccctcttc gacttcaaac aaggcatctt gggcggtggc caattcgctg aggactggct
     1261 ggacagcccg gccctgggcc ctggctggaa gcgccgcgaa gtctttcgca agtcaggggc
     1321 caccgcggga cgctcagaca cctattacca gagccccaca ggagacagga tccgaagcaa
     1381 agttgagctg actcgatacc tgggccctgc gggcgatctc accctcttcg acttcaaaca
     1441 aggcatcttg ggcggtggcc aattcgtacc ggtcgccacc atggtgagca agggcgagga
     1501 ggataacatg gccatcatca aggagttcat gcgcttcaag gtgcacatgg agggctccgt
     1561 gaacggccac gagttcgaga tcgagggcga gggcgagggc cgcccctacg agggcaccca
     1621 gaccgccaag ctgaaggtga ccaagggtgg ccccctgccc ttcgcctggg acatcctgtc
     1681 ccctcagttc atgtacggct ccaaggccta cgtgaagcac cccgccgaca tccccgacta
     1741 cttgaagctg tccttccccg agggcttcaa gtgggagcgc gtgatgaact tcgaggacgg
     1801 cggcgtggtg accgtgaccc aggactcctc cctgcaggac ggcgagttca tctacaaggt
     1861 gaagctgcgc ggcaccaact tcccctccga cggccccgta atgcagaaga agaccatggg
     1921 ctgggaggcc tcctccgagc ggatgtaccc cgaggacggc gccctgaagg gcgagatcaa
     1981 gcagaggctg aagctgaagg acggcggcca ctacgacgct gaggtcaaga ccacctacaa
     2041 ggccaagaag cccgtgcagc tgcccggcgc ctacaacgtc aacatcaagt tggacatcac
     2101 ctcccacaac gaggactaca ccatcgtgga acagtacgaa cgcgccgagg gccgccactc
     2161 caccggcggc atggacgagc tgtacaagta atcctctaga gtcgacctgc agaagcttgg
     2221 ccgccatggc ccaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc
     2281 acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc
     2341 atcaatgtat cttatcatgt ctggatcggg aattaattcg gcgcagcacc atggcctgaa
     2401 ataacctctg aaagaggaac ttggttaggt accttctgag gcggaaagaa ccagctgtgg
     2461 aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa
     2521 agcatgcatc tcaattagtc agcaaccagg tgtggaaagt ccccaggctc cccagcaggc
     2581 agaagtatgc aaagcatgca tctcaattag tcagcaacca tagtcccgcc cctaactccg
     2641 cccatcccgc ccctaactcc gcccagttcc gcccattctc cgccccatgg ctgactaatt
     2701 ttttttattt atgcagaggc cgaggccgcc tcggcctctg agctattcca gaagtagtga
     2761 ggaggctttt ttggaggcct aggcttttgc aaaaagctgt taacagcttg gcactggccg
     2821 tcgttttaca acgtcgtgac tgggaaaacc ctggcgttac ccaacttaat cgccttgcag
     2881 cacatccccc tttcgccagc tggcgtaata gcgaagaggc ccgcaccgat cgcccttccc
     2941 aacagttgcg cagcctgaat ggcgaatggc gcctgatgcg gtattttctc cttacgcatc
     3001 tgtgcggtat ttcacaccgc atacgtcaaa gcaaccatag tacgcgccct gtagcggcgc
     3061 attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgccct
     3121 agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg
     3181 tcaagctcta aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga
     3241 ccccaaaaaa cttgatttgg gtgatggttc acgtagtggg ccatcgccct gatagacggt
     3301 ttttcgccct ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg
     3361 aacaacactc aaccctatct cgggctattc ttttgattta taagggattt tgccgatttc
     3421 ggcctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt ttaacaaaat
     3481 attaacgttt acaattttat ggtgcactct cagtacaatc tgctctgatg ccgcatagtt
     3541 aagccagccc cgacacccgc caacacccgc tgacgcgccc tgacgggctt gtctgctccc
     3601 ggcatccgct tacagacaag ctgtgaccgt ctccgggagc tgcatgtgtc agaggttttc
     3661 accgtcatca ccgaaacgcg cgagacgaaa gggcctcgtg atacgcctat ttttataggt
     3721 taatgtcatg ataataatgg tttcttagac gtcaggtggc acttttcggg gaaatgtgcg
     3781 cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca
     3841 ataaccctga taaatgcttc aataatattg aaaaaggaag agtatgagta ttcaacattt
     3901 ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg ctcacccaga
     3961 aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg gttacatcga
     4021 actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac gttttccaat
     4081 gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtattg acgccgggca
     4141 agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt actcaccagt
     4201 cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg ctgccataac
     4261 catgagtgat aacactgcgg ccaacttact tctgacaacg atcggaggac cgaaggagct
     4321 aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt gggaaccgga
     4381 gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgtag caatggcaac
     4441 aacgttgcgc aaactattaa ctggcgaact acttactcta gcttcccggc aacaattaat
     4501 agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc ttccggctgg
     4561 ctggtttatt gctgataaat ctggagccgg tgagcgtggg tctcgcggta tcattgcagc
     4621 actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg ggagtcaggc
     4681 aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga ttaagcattg
     4741 gtaactgtca gaccaagttt actcatatat actttagatt gatttaaaac ttcattttta
     4801 atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg
     4861 tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga
     4921 tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt
     4981 ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag
     5041 agcgcagata ccaaatactg ttcttctagt gtagccgtag ttaggccacc acttcaagaa
     5101 ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag
     5161 tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca
     5221 gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac
     5281 cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa
     5341 ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc
     5401 agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg
     5461 tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc
     5521 ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgttctttc ctgcgttatc
     5581 ccctgattct gtggataacc gtattaccgc ctttgagtga gctgataccg ctcgccgcag
     5641 ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg gaagagcgcc caatacgcaa
     5701 accgcctctc cccgcgcgtt ggccgattca ttaatgcagc tggcacgaca ggtttcccga
     5761 ctggaaagcg ggcagtgagc gcaacgcaat taatgtgagt tagctcactc attaggcacc
     5821 ccaggcttta cactttatgc ttccggctcg tatgttgtgt ggaattgtga gcggataaca
     5881 atttcacaca ggaaacagct atgacatgat tacgaattaa ttcgagctcg cccgacat
//
