LOCUS       sequence-207356-        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        61..364
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        365..568
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     519..539
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      671..680
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1595..1615
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=7xHis
                     /translation="HHHHHHH"
     regulatory      1970..1979
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1976..1978,1979..1981,1982..2695)
                     /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(1976..1978,1979..1981,1982..2692)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(2021..2042)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2282..2301)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2629..2650
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    2818..2939
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2855..2874)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2909..2928
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2946..3401)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3083..3104)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3295..3314
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3428..3532
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3534..3891
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3534..3725
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3557..3577)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3742..3877
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3926..4720
                     /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             3926..4720
                     /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(3980..3999)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4590..4609
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4908..4927)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4952..4999
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5328..5916
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5817..5836
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg
       61 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
      121 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      181 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      241 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      301 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      361 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      421 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      481 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      541 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgcta
      601 ccggactcag atctcgagct tcgaattctg caattggtcg acggtaccgc gggcccggga
      661 tccaccggtc gccaccatgg tagctgggat gttagggctc agggaggaaa agccagcagt
      721 gccagaagcc tcggcttccc ccaaacagcg gcgctccatt atccgtgacc ggggacctat
      781 gtatgatgac cccaccttgc ctgaaggttg gacacgaaag ctcaaacaaa ggaagtctgg
      841 ccgatctgct ggaaagtatg atgtatattt gatcaatccc cagggaaaag cttttcgctc
      901 taaagtagaa ttgattgcat actttgaaaa ggtgggagac acctccttgg accctaatga
      961 ttttgacttc acggtaactg ggagagggag cccctccagg agagagcaga aaccacctaa
     1021 gaagcccaaa tctcccaaag ctccaggaac tggcaggggt cggggacgcc ccaaagggag
     1081 cggcactggg agaccaaagg cagcagcatc agaaggtgtt caggtgaaaa gggtcctgga
     1141 gaagagccct gggaaacttg ttgtcaagat gcctttccaa gcatcgcctg ggggtaaggg
     1201 tgagggaggt ggggctacca catctgccca ggtcatggtg atcaaacgcc ctggcagaaa
     1261 gcgaaaagct gaagctgacc cccaggccat tcctaagaaa cggggtagaa agcctgggag
     1321 tgtggtggca gctgctgcag ctgaggccaa aaagaaagcc gtgaaggagt cttccatacg
     1381 gtctgtgcat gagactgtgc tccccatcaa gaagcgcaag acccgggaga cggtcagcat
     1441 cgaggtcaag gaagtggtga agcccctgct ggtgtccacc cttggtgaga aaagcgggaa
     1501 gggactgaag acctgcaaga gccctgggcg taaaagcaag gagagcagcc ccaaggggcg
     1561 cagcagcagt gcctcctccc cacctaagaa ggagcaccat catcaccacc atcactcaga
     1621 gtccacaaag gcccccatgc cactgctccc atccccaccc ccacctgagc ctgagagctc
     1681 tgaggacccc atcagccccc ctgagcctca ggacttgagc agcagcatct gcaaagaaga
     1741 gaagatgccc cgaggaggct cactggaaag cgatggctgc cccaaggagc cagctaagac
     1801 tcagcctatg gtcgccacca ctaccacagt tgcagaaaag tacaaacacc gaggggaggg
     1861 agagcgcaaa gacattgttt catcttccat gccaaggcca aacagagagg agcctgtgga
     1921 cagccggacg cccgtgaccg agagagttag ctgtaaggat ccaccggtcg ccaccatggt
     1981 gagcaagggc gaggagctgt tcaccggggt ggtgcccatc ctggtcgagc tggacggcga
     2041 cgtaaacggc cacaagttca gcgtgtccgg cgagggcgag ggcgatgcca cctacggcaa
     2101 gctgaccctg aagttcatct gcaccaccgg caagctgccc gtgccctggc ccaccctcgt
     2161 gaccaccctg acctacggcg tgcagtgctt cagccgctac cccgaccaca tgaagcagca
     2221 cgacttcttc aagtccgcca tgcccgaagg ctacgtccag gagcgcacca tcttcttcaa
     2281 ggacgacggc aactacaaga cccgcgccga ggtgaagttc gagggcgaca ccctggtgaa
     2341 ccgcatcgag ctgaagggca tcgacttcaa ggaggacggc aacatcctgg ggcacaagct
     2401 ggagtacaac tacaacagcc acaacgtcta tatcatggcc gacaagcaga agaacggcat
     2461 caaggtgaac ttcaagatcc gccacaacat cgaggacggc agcgtgcagc tcgccgacca
     2521 ctaccagcag aacaccccca tcggcgacgg ccccgtgctg ctgcccgaca accactacct
     2581 gagcacccag tccgccctga gcaaagaccc caacgagaag cgcgatcaca tggtcctgct
     2641 ggagttcgtg accgccgccg ggatcactct cggcatggac gagctgtaca agtaaagcgg
     2701 ccgcgactct agatcataat cagccatacc acatttgtag aggttttact tgctttaaaa
     2761 aacctcccac acctccccct gaacctgaaa cataaaatga atgcaattgt tgttgttaac
     2821 ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat
     2881 aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttaa
     2941 ggcgtaaatt gtaagcgtta atattttgtt aaaattcgcg ttaaattttt gttaaatcag
     3001 ctcatttttt aaccaatagg ccgaaatcgg caaaatccct tataaatcaa aagaatagac
     3061 cgagataggg ttgagtgttg ttccagtttg gaacaagagt ccactattaa agaacgtgga
     3121 ctccaacgtc aaagggcgaa aaaccgtcta tcagggcgat ggcccactac gtgaaccatc
     3181 accctaatca agttttttgg ggtcgaggtg ccgtaaagca ctaaatcgga accctaaagg
     3241 gagcccccga tttagagctt gacggggaaa gccggcgaac gtggcgagaa aggaagggaa
     3301 gaaagcgaaa ggagcgggcg ctagggcgct ggcaagtgta gcggtcacgc tgcgcgtaac
     3361 caccacaccc gccgcgctta atgcgccgct acagggcgcg tcaggtggca cttttcgggg
     3421 aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct
     3481 catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga gtcctgaggc
     3541 ggaaagaacc agctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca
     3601 gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccaggtg tggaaagtcc
     3661 ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccata
     3721 gtcccgcccc taactccgcc catcccgccc ctaactccgc ccagttccgc ccattctccg
     3781 ccccatggct gactaatttt ttttatttat gcagaggccg aggccgcctc ggcctctgag
     3841 ctattccaga agtagtgagg aggctttttt ggaggcctag gcttttgcaa agatcgatca
     3901 agagacagga tgaggatcgt ttcgcatgat tgaacaagat ggattgcacg caggttctcc
     3961 ggccgcttgg gtggagaggc tattcggcta tgactgggca caacagacaa tcggctgctc
     4021 tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg gttctttttg tcaagaccga
     4081 cctgtccggt gccctgaatg aactgcaaga cgaggcagcg cggctatcgt ggctggccac
     4141 gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact gaagcgggaa gggactggct
     4201 gctattgggc gaagtgccgg ggcaggatct cctgtcatct caccttgctc ctgccgagaa
     4261 agtatccatc atggctgatg caatgcggcg gctgcatacg cttgatccgg ctacctgccc
     4321 attcgaccac caagcgaaac atcgcatcga gcgagcacgt actcggatgg aagccggtct
     4381 tgtcgatcag gatgatctgg acgaagagca tcaggggctc gcgccagccg aactgttcgc
     4441 caggctcaag gcgagcatgc ccgacggcga ggatctcgtc gtgacccatg gcgatgcctg
     4501 cttgccgaat atcatggtgg aaaatggccg cttttctgga ttcatcgact gtggccggct
     4561 gggtgtggcg gaccgctatc aggacatagc gttggctacc cgtgatattg ctgaagagct
     4621 tggcggcgaa tgggctgacc gcttcctcgt gctttacggt atcgccgctc ccgattcgca
     4681 gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga gcgggactct ggggttcgaa
     4741 atgaccgacc aagcgacgcc caacctgcca tcacgagatt tcgattccac cgccgccttc
     4801 tatgaaaggt tgggcttcgg aatcgttttc cgggacgccg gctggatgat cctccagcgc
     4861 ggggatctca tgctggagtt cttcgcccac cctaggggga ggctaactga aacacggaag
     4921 gagacaatac cggaaggaac ccgcgctatg acggcaataa aaagacagaa taaaacgcac
     4981 ggtgttgggt cgtttgttca taaacgcggg gttcggtccc agggctggca ctctgtcgat
     5041 accccaccga gaccccattg gggccaatac gcccgcgttt cttccttttc cccaccccac
     5101 cccccaagtt cgggtgaagg cccagggctc gcagccaacg tcggggcggc aggccctgcc
     5161 atagcctcag gttactcata tatactttag attgatttaa aacttcattt ttaatttaaa
     5221 aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt
     5281 tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt
     5341 tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt
     5401 ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag
     5461 ataccaaata ctgtccttct agtgtagccg tagttaggcc accacttcaa gaactctgta
     5521 gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat
     5581 aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg
     5641 ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg
     5701 agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac
     5761 aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga
     5821 aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt
     5881 ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta
     5941 cggttcctgg ccttttgctg gccttttgct cacatgttct ttcctgcgtt atcccctgat
     6001 tctgtggata accgtattac cgccatgcat
//
