LOCUS       sequence-210788-        6149 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of mouse MeCP2 (R111G) in mammalian cells .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6149)
  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 6149)
  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(R111G)
FEATURES             Location/Qualifiers
     source          1..6149
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        51..354
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        355..558
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     509..529
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             1704..1724
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=7xHis
                     /translation="HHHHHHH"
     regulatory      2079..2088
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(2085..2087,2088..2090,2091..2804)
                     /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(2085..2087,2088..2090,2091..2801)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(2130..2151)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2391..2410)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2738..2759
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    2927..3048
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2964..2983)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3018..3037
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3055..3510)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3192..3213)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3404..3423
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3537..3641
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3643..4000
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3643..3834
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3666..3686)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3851..3986
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4035..4829
                     /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             4035..4829
                     /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(4089..4108)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4699..4718
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5017..5036)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5061..5108
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5437..6025
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5926..5945
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg cgttacataa
       61 cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt gacgtcaata
      121 atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca atgggtggag
      181 tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc aagtacgccc
      241 cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta catgacctta
      301 tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac catggtgatg
      361 cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg atttccaagt
      421 ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg ggactttcca
      481 aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt acggtgggag
      541 gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgcta ccggactcag
      601 atctcgagat ggtagctggg atgttagggc tcagggagga aaagtcagaa gaccaggatc
      661 tccagggcct cagagacaag ccactgaagt ttaagaaggc gaagaaagac aagaaggagg
      721 acaaagaagg caagcatgag ccactacaac cttcagccca ccattctgca gagccagcag
      781 aggcaggcaa agcagaaaca tcagaaagct caggctctgc cccagcagtg ccagaagcct
      841 cggcttcccc caaacagcgg cgctccatta tccgtgaccg gggacctatg tatgatgacc
      901 ccaccttgcc tgaaggttgg acacgaaagc ttaaacaagg gaagtctggc cgatctgctg
      961 gaaagtatga tgtatatttg atcaatcccc agggaaaagc ttttcgctct aaagtagaat
     1021 tgattgcata ctttgaaaag gtgggagaca cctccttgga ccctaatgat tttgacttca
     1081 cggtaactgg gagagggagc ccctccagga gagagcagaa accacctaag aagcccaaat
     1141 ctcccaaagc tccaggaact ggcaggggtc ggggacgccc caaagggagc ggcactggga
     1201 gaccaaaggc agcagcatca gaaggtgttc aggtgaaaag ggtcctggag aagagccctg
     1261 ggaaacttgt tgtcaagatg cctttccaag catcgcctgg gggtaagggt gagggaggtg
     1321 gggctaccac atctgcccag gtcatggtga tcaaacgccc tggcagaaag cgaaaagctg
     1381 aagctgaccc ccaggccatt cctaagaaac ggggtagaaa gcctgggagt gtggtggcag
     1441 ctgctgcagc tgaggccaaa aagaaagccg tgaaggagtc ttccatacgg tctgtgcatg
     1501 agactgtgct ccccatcaag aagcgcaaga cccgggagac ggtcagcatc gaggtcaagg
     1561 aagtggtgaa gcccctgctg gtgtccaccc ttggtgagaa aagcgggaag ggactgaaga
     1621 cctgcaagag ccctgggcgt aaaagcaagg agagcagccc caaggggcgc agcagcagtg
     1681 cctcctcccc acctaagaag gagcaccatc atcaccacca tcactcagag tccacaaagg
     1741 cccccatgcc actgctccca tccccacccc cacctgagcc tgagagctct gaggacccca
     1801 tcagcccccc tgagcctcag gacttgagca gcagcatctg caaagaagag aagatgcccc
     1861 gaggaggctc actggaaagc gatggctgcc ccaaggagcc agctaagact cagcctatgg
     1921 tcgccaccac taccacagtt gcagaaaagt acaaacaccg aggggaggga gagcgcaaag
     1981 acattgtttc atcttccatg ccaaggccaa acagagagga gcctgtggac agccggacgc
     2041 ccgtgaccga gagagttagc tgtaaggatc caccggtcgc caccatggtg agcaagggcg
     2101 aggagctgtt caccggggtg gtgcccatcc tggtcgagct ggacggcgac gtaaacggcc
     2161 acaagttcag cgtgtccggc gagggcgagg gcgatgccac ctacggcaag ctgaccctga
     2221 agttcatctg caccaccggc aagctgcccg tgccctggcc caccctcgtg accaccctga
     2281 cctacggcgt gcagtgcttc agccgctacc ccgaccacat gaagcagcac gacttcttca
     2341 agtccgccat gcccgaaggc tacgtccagg agcgcaccat cttcttcaag gacgacggca
     2401 actacaagac ccgcgccgag gtgaagttcg agggcgacac cctggtgaac cgcatcgagc
     2461 tgaagggcat cgacttcaag gaggacggca acatcctggg gcacaagctg gagtacaact
     2521 acaacagcca caacgtctat atcatggccg acaagcagaa gaacggcatc aaggtgaact
     2581 tcaagatccg ccacaacatc gaggacggca gcgtgcagct cgccgaccac taccagcaga
     2641 acacccccat cggcgacggc cccgtgctgc tgcccgacaa ccactacctg agcacccagt
     2701 ccgccctgag caaagacccc aacgagaagc gcgatcacat ggtcctgctg gagttcgtga
     2761 ccgccgccgg gatcactctc ggcatggacg agctgtacaa gtaaagcggc cgcgactcta
     2821 gatcataatc agccatacca catttgtaga ggttttactt gctttaaaaa acctcccaca
     2881 cctccccctg aacctgaaac ataaaatgaa tgcaattgtt gttgttaact tgtttattgc
     2941 agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt
     3001 ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttaag gcgtaaattg
     3061 taagcgttaa tattttgtta aaattcgcgt taaatttttg ttaaatcagc tcatttttta
     3121 accaataggc cgaaatcggc aaaatccctt ataaatcaaa agaatagacc gagatagggt
     3181 tgagtgttgt tccagtttgg aacaagagtc cactattaaa gaacgtggac tccaacgtca
     3241 aagggcgaaa aaccgtctat cagggcgatg gcccactacg tgaaccatca ccctaatcaa
     3301 gttttttggg gtcgaggtgc cgtaaagcac taaatcggaa ccctaaaggg agcccccgat
     3361 ttagagcttg acggggaaag ccggcgaacg tggcgagaaa ggaagggaag aaagcgaaag
     3421 gagcgggcgc tagggcgctg gcaagtgtag cggtcacgct gcgcgtaacc accacacccg
     3481 ccgcgcttaa tgcgccgcta cagggcgcgt caggtggcac ttttcgggga aatgtgcgcg
     3541 gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat
     3601 aaccctgata aatgcttcaa taatattgaa aaaggaagag tcctgaggcg gaaagaacca
     3661 gctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag caggcagaag
     3721 tatgcaaagc atgcatctca attagtcagc aaccaggtgt ggaaagtccc caggctcccc
     3781 agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccatag tcccgcccct
     3841 aactccgccc atcccgcccc taactccgcc cagttccgcc cattctccgc cccatggctg
     3901 actaattttt tttatttatg cagaggccga ggccgcctcg gcctctgagc tattccagaa
     3961 gtagtgagga ggcttttttg gaggcctagg cttttgcaaa gatcgatcaa gagacaggat
     4021 gaggatcgtt tcgcatgatt gaacaagatg gattgcacgc aggttctccg gccgcttggg
     4081 tggagaggct attcggctat gactgggcac aacagacaat cggctgctct gatgccgccg
     4141 tgttccggct gtcagcgcag gggcgcccgg ttctttttgt caagaccgac ctgtccggtg
     4201 ccctgaatga actgcaagac gaggcagcgc ggctatcgtg gctggccacg acgggcgttc
     4261 cttgcgcagc tgtgctcgac gttgtcactg aagcgggaag ggactggctg ctattgggcg
     4321 aagtgccggg gcaggatctc ctgtcatctc accttgctcc tgccgagaaa gtatccatca
     4381 tggctgatgc aatgcggcgg ctgcatacgc ttgatccggc tacctgccca ttcgaccacc
     4441 aagcgaaaca tcgcatcgag cgagcacgta ctcggatgga agccggtctt gtcgatcagg
     4501 atgatctgga cgaagagcat caggggctcg cgccagccga actgttcgcc aggctcaagg
     4561 cgagcatgcc cgacggcgag gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata
     4621 tcatggtgga aaatggccgc ttttctggat tcatcgactg tggccggctg ggtgtggcgg
     4681 accgctatca ggacatagcg ttggctaccc gtgatattgc tgaagagctt ggcggcgaat
     4741 gggctgaccg cttcctcgtg ctttacggta tcgccgctcc cgattcgcag cgcatcgcct
     4801 tctatcgcct tcttgacgag ttcttctgag cgggactctg gggttcgaaa tgaccgacca
     4861 agcgacgccc aacctgccat cacgagattt cgattccacc gccgccttct atgaaaggtt
     4921 gggcttcgga atcgttttcc gggacgccgg ctggatgatc ctccagcgcg gggatctcat
     4981 gctggagttc ttcgcccacc ctagggggag gctaactgaa acacggaagg agacaatacc
     5041 ggaaggaacc cgcgctatga cggcaataaa aagacagaat aaaacgcacg gtgttgggtc
     5101 gtttgttcat aaacgcgggg ttcggtccca gggctggcac tctgtcgata ccccaccgag
     5161 accccattgg ggccaatacg cccgcgtttc ttccttttcc ccaccccacc ccccaagttc
     5221 gggtgaaggc ccagggctcg cagccaacgt cggggcggca ggccctgcca tagcctcagg
     5281 ttactcatat atactttaga ttgatttaaa acttcatttt taatttaaaa ggatctaggt
     5341 gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg
     5401 agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt
     5461 aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca
     5521 agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac
     5581 tgttcttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac
     5641 atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct
     5701 taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg
     5761 gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga gatacctaca
     5821 gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt
     5881 aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta
     5941 tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc
     6001 gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc
     6061 cttttgctgg ccttttgctc acatgttctt tcctgcgtta tcccctgatt ctgtggataa
     6121 ccgtattacc gccatgcatt agttattaa
//
