LOCUS       sequence-210793-        6030 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of mouse MeCP2 (deltaN) in mammalian cells .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6030)
  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 6030)
  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(deltaN)
FEATURES             Location/Qualifiers
     source          1..6030
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        56..359
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        360..563
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     514..534
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      666..675
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1590..1610
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=7xHis
                     /translation="HHHHHHH"
     regulatory      1965..1974
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1971..1973,1974..1976,1977..2690)
                     /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(1971..1973,1974..1976,1977..2687)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(2016..2037)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2277..2296)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2624..2645
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    2813..2934
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2850..2869)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2904..2923
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2941..3396)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3078..3099)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3290..3309
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3423..3527
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3529..3886
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3529..3720
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3552..3572)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3737..3872
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3921..4715
                     /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             3921..4715
                     /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(3975..3994)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4585..4604
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4903..4922)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4947..4994
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5323..5911
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5812..5831
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 attaatagta atcaattacg gggtcattag ttcatagccc atatatggag ttccgcgtta
       61 cataacttac ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc ccattgacgt
      121 caataatgac gtatgttccc atagtaacgc caatagggac tttccattga cgtcaatggg
      181 tggagtattt acggtaaact gcccacttgg cagtacatca agtgtatcat atgccaagta
      241 cgccccctat tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc cagtacatga
      301 ccttatggga ctttcctact tggcagtaca tctacgtatt agtcatcgct attaccatgg
      361 tgatgcggtt ttggcagtac atcaatgggc gtggatagcg gtttgactca cggggatttc
      421 caagtctcca ccccattgac gtcaatggga gtttgttttg gcaccaaaat caacgggact
      481 ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat gggcggtagg cgtgtacggt
      541 gggaggtcta tataagcaga gctggtttag tgaaccgtca gatccgctag cgctaccgga
      601 ctcagatctc gagcttcgaa ttctgcaatt ggtcgacggt accgcgggcc cgggatccac
      661 cggtcgccac catggtagct gggatgttag ggctcaggga ggaaaagcca gcagtgccag
      721 aagcctcggc ttcccccaaa cagcggcgct ccattatccg tgaccgggga cctatgtatg
      781 atgaccccac cttgcctgaa ggttggacac gaaagctcaa acaaaggaag tctggccgat
      841 ctgctggaaa gtatgatgta tatttgatca atccccaggg aaaagctttt cgctctaaag
      901 tagaattgat tgcatacttt gaaaaggtgg gagacacctc cttggaccct aatgattttg
      961 acttcacggt aactgggaga gggagcccct ccaggagaga gcagaaacca cctaagaagc
     1021 ccaaatctcc caaagctcca ggaactggca ggggtcgggg acgccccaaa gggagcggca
     1081 ctgggagacc aaaggcagca gcatcagaag gtgttcaggt gaaaagggtc ctggagaaga
     1141 gccctgggaa acttgttgtc aagatgcctt tccaagcatc gcctgggggt aagggtgagg
     1201 gaggtggggc taccacatct gcccaggtca tggtgatcaa acgccctggc agaaagcgaa
     1261 aagctgaagc tgacccccag gccattccta agaaacgggg tagaaagcct gggagtgtgg
     1321 tggcagctgc tgcagctgag gccaaaaaga aagccgtgaa ggagtcttcc atacggtctg
     1381 tgcatgagac tgtgctcccc atcaagaagc gcaagacccg ggagacggtc agcatcgagg
     1441 tcaaggaagt ggtgaagccc ctgctggtgt ccacccttgg tgagaaaagc gggaagggac
     1501 tgaagacctg caagagccct gggcgtaaaa gcaaggagag cagccccaag gggcgcagca
     1561 gcagtgcctc ctccccacct aagaaggagc accatcatca ccaccatcac tcagagtcca
     1621 caaaggcccc catgccactg ctcccatccc cacccccacc tgagcctgag agctctgagg
     1681 accccatcag cccccctgag cctcaggact tgagcagcag catctgcaaa gaagagaaga
     1741 tgccccgagg aggctcactg gaaagcgatg gctgccccaa ggagccagct aagactcagc
     1801 ctatggtcgc caccactacc acagttgcag aaaagtacaa acaccgaggg gagggagagc
     1861 gcaaagacat tgtttcatct tccatgccaa ggccaaacag agaggagcct gtggacagcc
     1921 ggacgcccgt gaccgagaga gttagctgta aggatccacc ggtcgccacc atggtgagca
     1981 agggcgagga gctgttcacc ggggtggtgc ccatcctggt cgagctggac ggcgacgtaa
     2041 acggccacaa gttcagcgtg tccggcgagg gcgagggcga tgccacctac ggcaagctga
     2101 ccctgaagtt catctgcacc accggcaagc tgcccgtgcc ctggcccacc ctcgtgacca
     2161 ccctgaccta cggcgtgcag tgcttcagcc gctaccccga ccacatgaag cagcacgact
     2221 tcttcaagtc cgccatgccc gaaggctacg tccaggagcg caccatcttc ttcaaggacg
     2281 acggcaacta caagacccgc gccgaggtga agttcgaggg cgacaccctg gtgaaccgca
     2341 tcgagctgaa gggcatcgac ttcaaggagg acggcaacat cctggggcac aagctggagt
     2401 acaactacaa cagccacaac gtctatatca tggccgacaa gcagaagaac ggcatcaagg
     2461 tgaacttcaa gatccgccac aacatcgagg acggcagcgt gcagctcgcc gaccactacc
     2521 agcagaacac ccccatcggc gacggccccg tgctgctgcc cgacaaccac tacctgagca
     2581 cccagtccgc cctgagcaaa gaccccaacg agaagcgcga tcacatggtc ctgctggagt
     2641 tcgtgaccgc cgccgggatc actctcggca tggacgagct gtacaagtaa agcggccgcg
     2701 actctagatc ataatcagcc ataccacatt tgtagaggtt ttacttgctt taaaaaacct
     2761 cccacacctc cccctgaacc tgaaacataa aatgaatgca attgttgttg ttaacttgtt
     2821 tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc
     2881 atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttaaggcgt
     2941 aaattgtaag cgttaatatt ttgttaaaat tcgcgttaaa tttttgttaa atcagctcat
     3001 tttttaacca ataggccgaa atcggcaaaa tcccttataa atcaaaagaa tagaccgaga
     3061 tagggttgag tgttgttcca gtttggaaca agagtccact attaaagaac gtggactcca
     3121 acgtcaaagg gcgaaaaacc gtctatcagg gcgatggccc actacgtgaa ccatcaccct
     3181 aatcaagttt tttggggtcg aggtgccgta aagcactaaa tcggaaccct aaagggagcc
     3241 cccgatttag agcttgacgg ggaaagccgg cgaacgtggc gagaaaggaa gggaagaaag
     3301 cgaaaggagc gggcgctagg gcgctggcaa gtgtagcggt cacgctgcgc gtaaccacca
     3361 cacccgccgc gcttaatgcg ccgctacagg gcgcgtcagg tggcactttt cggggaaatg
     3421 tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga
     3481 gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtcct gaggcggaaa
     3541 gaaccagctg tggaatgtgt gtcagttagg gtgtggaaag tccccaggct ccccagcagg
     3601 cagaagtatg caaagcatgc atctcaatta gtcagcaacc aggtgtggaa agtccccagg
     3661 ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc
     3721 gcccctaact ccgcccatcc cgcccctaac tccgcccagt tccgcccatt ctccgcccca
     3781 tggctgacta atttttttta tttatgcaga ggccgaggcc gcctcggcct ctgagctatt
     3841 ccagaagtag tgaggaggct tttttggagg cctaggcttt tgcaaagatc gatcaagaga
     3901 caggatgagg atcgtttcgc atgattgaac aagatggatt gcacgcaggt tctccggccg
     3961 cttgggtgga gaggctattc ggctatgact gggcacaaca gacaatcggc tgctctgatg
     4021 ccgccgtgtt ccggctgtca gcgcaggggc gcccggttct ttttgtcaag accgacctgt
     4081 ccggtgccct gaatgaactg caagacgagg cagcgcggct atcgtggctg gccacgacgg
     4141 gcgttccttg cgcagctgtg ctcgacgttg tcactgaagc gggaagggac tggctgctat
     4201 tgggcgaagt gccggggcag gatctcctgt catctcacct tgctcctgcc gagaaagtat
     4261 ccatcatggc tgatgcaatg cggcggctgc atacgcttga tccggctacc tgcccattcg
     4321 accaccaagc gaaacatcgc atcgagcgag cacgtactcg gatggaagcc ggtcttgtcg
     4381 atcaggatga tctggacgaa gagcatcagg ggctcgcgcc agccgaactg ttcgccaggc
     4441 tcaaggcgag catgcccgac ggcgaggatc tcgtcgtgac ccatggcgat gcctgcttgc
     4501 cgaatatcat ggtggaaaat ggccgctttt ctggattcat cgactgtggc cggctgggtg
     4561 tggcggaccg ctatcaggac atagcgttgg ctacccgtga tattgctgaa gagcttggcg
     4621 gcgaatgggc tgaccgcttc ctcgtgcttt acggtatcgc cgctcccgat tcgcagcgca
     4681 tcgccttcta tcgccttctt gacgagttct tctgagcggg actctggggt tcgaaatgac
     4741 cgaccaagcg acgcccaacc tgccatcacg agatttcgat tccaccgccg ccttctatga
     4801 aaggttgggc ttcggaatcg ttttccggga cgccggctgg atgatcctcc agcgcgggga
     4861 tctcatgctg gagttcttcg cccaccctag ggggaggcta actgaaacac ggaaggagac
     4921 aataccggaa ggaacccgcg ctatgacggc aataaaaaga cagaataaaa cgcacggtgt
     4981 tgggtcgttt gttcataaac gcggggttcg gtcccagggc tggcactctg tcgatacccc
     5041 accgagaccc cattggggcc aatacgcccg cgtttcttcc ttttccccac cccacccccc
     5101 aagttcgggt gaaggcccag ggctcgcagc caacgtcggg gcggcaggcc ctgccatagc
     5161 ctcaggttac tcatatatac tttagattga tttaaaactt catttttaat ttaaaaggat
     5221 ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt
     5281 ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct
     5341 gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc
     5401 ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc
     5461 aaatactgtt cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc
     5521 gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc
     5581 gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg
     5641 aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata
     5701 cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta
     5761 tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc
     5821 ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg
     5881 atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt
     5941 cctggccttt tgctggcctt ttgctcacat gttctttcct gcgttatccc ctgattctgt
     6001 ggataaccgt attaccgcca tgcattagtt
//
