LOCUS       sequence-210797-        5511 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of mouse MeCP2 (deltaNC) in mammalian cells .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5511)
  AUTHORS   Tillotson R, Selfridge J, Koerner MV, Gadalla KKE, Guy J, De Sousa 
            D, Hector RD, Cobb SR, Bird A
  TITLE     Radically truncated MeCP2 rescues Rett syndrome-like neurological 
            defects.
  JOURNAL   Nature. 2017 Oct 19;550(7676):398-401. doi: 10.1038/nature24058. 
            Epub 2017 Oct 11.
  PUBMED    29019980
REFERENCE   2  (bases 1 to 5511)
  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: 
            "10.1038/nature24058"; journalName: "Nature"; date: "2017-10-19- 
            19"; volume: "550"; issue: "7676"; pages: "398-401"
COMMENT     Sequence Label: pEGFP-N1_MeCP2(deltaNC)
FEATURES             Location/Qualifiers
     source          1..5511
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        55..358
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        359..562
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     513..533
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      665..674
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1451..1453,1454..1456,1457..2170)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(1451..1453,1454..1456,1457..2167)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1496..1517)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1757..1776)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2104..2125
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    2293..2414
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2330..2349)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2384..2403
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2421..2876)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2558..2579)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2770..2789
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2903..3007
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3009..3366
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3009..3200
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3032..3052)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3217..3352
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3401..4195
                     /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             3401..4195
                     /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(3455..3474)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4065..4084
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4383..4402)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4427..4474
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4803..5391
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5292..5311
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ttaatagtaa tcaattacgg ggtcattagt tcatagccca tatatggagt tccgcgttac
       61 ataacttacg gtaaatggcc cgcctggctg accgcccaac gacccccgcc cattgacgtc
      121 aataatgacg tatgttccca tagtaacgcc aatagggact ttccattgac gtcaatgggt
      181 ggagtattta cggtaaactg cccacttggc agtacatcaa gtgtatcata tgccaagtac
      241 gccccctatt gacgtcaatg acggtaaatg gcccgcctgg cattatgccc agtacatgac
      301 cttatgggac tttcctactt ggcagtacat ctacgtatta gtcatcgcta ttaccatggt
      361 gatgcggttt tggcagtaca tcaatgggcg tggatagcgg tttgactcac ggggatttcc
      421 aagtctccac cccattgacg tcaatgggag tttgttttgg caccaaaatc aacgggactt
      481 tccaaaatgt cgtaacaact ccgccccatt gacgcaaatg ggcggtaggc gtgtacggtg
      541 ggaggtctat ataagcagag ctggtttagt gaaccgtcag atccgctagc gctaccggac
      601 tcagatctcg agcttcgaat tctgcaattg gtcgacggta ccgcgggccc gggatccacc
      661 ggtcgccacc atggtagctg ggatgttagg gctcagggag gaaaagccag cagtgccaga
      721 agcctcggct tcccccaaac agcggcgctc cattatccgt gaccggggac ctatgtatga
      781 tgaccccacc ttgcctgaag gttggacacg aaagctcaaa caaaggaagt ctggccgatc
      841 tgctggaaag tatgatgtat atttgatcaa tccccaggga aaagcttttc gctctaaagt
      901 agaattgatt gcatactttg aaaaggtggg agacacctcc ttggacccta atgattttga
      961 cttcacggta actgggagag ggagcccctc caggagagag cagaaaccac ctaagaagcc
     1021 caaatctccc aaagctccag gaactggcag gggtcgggga cgccccaaag ggagcggcac
     1081 tgggagacca aaggcagcag catcagaagg tgttcaggtg aaaagggtcc tggagaagag
     1141 ccctgggaaa cttgttgtca agatgccttt ccaagcatcg cctgggggta agggtgaggg
     1201 aggtggggct accacatctg cccaggtcat ggtgatcaaa cgccctggca gaaagcgaaa
     1261 agctgaagct gacccccagg ccattcctaa gaaacggggt agaaagcctg ggagtgtggt
     1321 ggcagctgct gcagctgagg ccaaaaagaa agccgtgaag gagtcttcca tacggtctgt
     1381 gcatgagact gtgctcccca tcaagaagcg caagacccgg gagacggtcg ggagctccgg
     1441 cagttctgga atggtgagca agggcgagga gctgttcacc ggggtggtgc ccatcctggt
     1501 cgagctggac ggcgacgtaa acggccacaa gttcagcgtg tccggcgagg gcgagggcga
     1561 tgccacctac ggcaagctga ccctgaagtt catctgcacc accggcaagc tgcccgtgcc
     1621 ctggcccacc ctcgtgacca ccctgaccta cggcgtgcag tgcttcagcc gctaccccga
     1681 ccacatgaag cagcacgact tcttcaagtc cgccatgccc gaaggctacg tccaggagcg
     1741 caccatcttc ttcaaggacg acggcaacta caagacccgc gccgaggtga agttcgaggg
     1801 cgacaccctg gtgaaccgca tcgagctgaa gggcatcgac ttcaaggagg acggcaacat
     1861 cctggggcac aagctggagt acaactacaa cagccacaac gtctatatca tggccgacaa
     1921 gcagaagaac ggcatcaagg tgaacttcaa gatccgccac aacatcgagg acggcagcgt
     1981 gcagctcgcc gaccactacc agcagaacac ccccatcggc gacggccccg tgctgctgcc
     2041 cgacaaccac tacctgagca cccagtccgc cctgagcaaa gaccccaacg agaagcgcga
     2101 tcacatggtc ctgctggagt tcgtgaccgc cgccgggatc actctcggca tggacgagct
     2161 gtacaagtaa agcggccgcg actctagatc ataatcagcc ataccacatt tgtagaggtt
     2221 ttacttgctt taaaaaacct cccacacctc cccctgaacc tgaaacataa aatgaatgca
     2281 attgttgttg ttaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc
     2341 acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc
     2401 atcaatgtat cttaaggcgt aaattgtaag cgttaatatt ttgttaaaat tcgcgttaaa
     2461 tttttgttaa atcagctcat tttttaacca ataggccgaa atcggcaaaa tcccttataa
     2521 atcaaaagaa tagaccgaga tagggttgag tgttgttcca gtttggaaca agagtccact
     2581 attaaagaac gtggactcca acgtcaaagg gcgaaaaacc gtctatcagg gcgatggccc
     2641 actacgtgaa ccatcaccct aatcaagttt tttggggtcg aggtgccgta aagcactaaa
     2701 tcggaaccct aaagggagcc cccgatttag agcttgacgg ggaaagccgg cgaacgtggc
     2761 gagaaaggaa gggaagaaag cgaaaggagc gggcgctagg gcgctggcaa gtgtagcggt
     2821 cacgctgcgc gtaaccacca cacccgccgc gcttaatgcg ccgctacagg gcgcgtcagg
     2881 tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc
     2941 aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag
     3001 gaagagtcct gaggcggaaa gaaccagctg tggaatgtgt gtcagttagg gtgtggaaag
     3061 tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc
     3121 aggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat
     3181 tagtcagcaa ccatagtccc gcccctaact ccgcccatcc cgcccctaac tccgcccagt
     3241 tccgcccatt ctccgcccca tggctgacta atttttttta tttatgcaga ggccgaggcc
     3301 gcctcggcct ctgagctatt ccagaagtag tgaggaggct tttttggagg cctaggcttt
     3361 tgcaaagatc gatcaagaga caggatgagg atcgtttcgc atgattgaac aagatggatt
     3421 gcacgcaggt tctccggccg cttgggtgga gaggctattc ggctatgact gggcacaaca
     3481 gacaatcggc tgctctgatg ccgccgtgtt ccggctgtca gcgcaggggc gcccggttct
     3541 ttttgtcaag accgacctgt ccggtgccct gaatgaactg caagacgagg cagcgcggct
     3601 atcgtggctg gccacgacgg gcgttccttg cgcagctgtg ctcgacgttg tcactgaagc
     3661 gggaagggac tggctgctat tgggcgaagt gccggggcag gatctcctgt catctcacct
     3721 tgctcctgcc gagaaagtat ccatcatggc tgatgcaatg cggcggctgc atacgcttga
     3781 tccggctacc tgcccattcg accaccaagc gaaacatcgc atcgagcgag cacgtactcg
     3841 gatggaagcc ggtcttgtcg atcaggatga tctggacgaa gagcatcagg ggctcgcgcc
     3901 agccgaactg ttcgccaggc tcaaggcgag catgcccgac ggcgaggatc tcgtcgtgac
     3961 ccatggcgat gcctgcttgc cgaatatcat ggtggaaaat ggccgctttt ctggattcat
     4021 cgactgtggc cggctgggtg tggcggaccg ctatcaggac atagcgttgg ctacccgtga
     4081 tattgctgaa gagcttggcg gcgaatgggc tgaccgcttc ctcgtgcttt acggtatcgc
     4141 cgctcccgat tcgcagcgca tcgccttcta tcgccttctt gacgagttct tctgagcggg
     4201 actctggggt tcgaaatgac cgaccaagcg acgcccaacc tgccatcacg agatttcgat
     4261 tccaccgccg ccttctatga aaggttgggc ttcggaatcg ttttccggga cgccggctgg
     4321 atgatcctcc agcgcgggga tctcatgctg gagttcttcg cccaccctag ggggaggcta
     4381 actgaaacac ggaaggagac aataccggaa ggaacccgcg ctatgacggc aataaaaaga
     4441 cagaataaaa cgcacggtgt tgggtcgttt gttcataaac gcggggttcg gtcccagggc
     4501 tggcactctg tcgatacccc accgagaccc cattggggcc aatacgcccg cgtttcttcc
     4561 ttttccccac cccacccccc aagttcgggt gaaggcccag ggctcgcagc caacgtcggg
     4621 gcggcaggcc ctgccatagc ctcaggttac tcatatatac tttagattga tttaaaactt
     4681 catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc
     4741 ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct
     4801 tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta
     4861 ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc
     4921 ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt aggccaccac
     4981 ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct
     5041 gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat
     5101 aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg
     5161 acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa
     5221 gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg
     5281 gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga
     5341 cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc
     5401 aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct
     5461 gcgttatccc ctgattctgt ggataaccgt attaccgcca tgcattagtt a
//
