LOCUS       sequence-210786-        6149 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of mouse MeCP2 (WT) 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(WT)
FEATURES             Location/Qualifiers
     source          1..6149
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        58..361
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        362..565
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     516..536
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             1711..1731
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=7xHis
                     /translation="HHHHHHH"
     regulatory      2086..2095
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(2092..2094,2095..2097,2098..2811)
                     /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(2092..2094,2095..2097,2098..2808)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(2137..2158)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2398..2417)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2745..2766
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    2934..3055
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2971..2990)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3025..3044
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3062..3517)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3199..3220)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3411..3430
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3544..3648
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3650..4007
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3650..3841
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3673..3693)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3858..3993
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4042..4836
                     /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             4042..4836
                     /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(4096..4115)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4706..4725
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5024..5043)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5068..5115
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5444..6032
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5933..5952
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ttattaatag taatcaatta cggggtcatt agttcatagc ccatatatgg agttccgcgt
       61 tacataactt acggtaaatg gcccgcctgg ctgaccgccc aacgaccccc gcccattgac
      121 gtcaataatg acgtatgttc ccatagtaac gccaataggg actttccatt gacgtcaatg
      181 ggtggagtat ttacggtaaa ctgcccactt ggcagtacat caagtgtatc atatgccaag
      241 tacgccccct attgacgtca atgacggtaa atggcccgcc tggcattatg cccagtacat
      301 gaccttatgg gactttccta cttggcagta catctacgta ttagtcatcg ctattaccat
      361 ggtgatgcgg ttttggcagt acatcaatgg gcgtggatag cggtttgact cacggggatt
      421 tccaagtctc caccccattg acgtcaatgg gagtttgttt tggcaccaaa atcaacggga
      481 ctttccaaaa tgtcgtaaca actccgcccc attgacgcaa atgggcggta ggcgtgtacg
      541 gtgggaggtc tatataagca gagctggttt agtgaaccgt cagatccgct agcgctaccg
      601 gactcagatc tcgagatggt agctgggatg ttagggctca gggaggaaaa gtcagaagac
      661 caggatctcc agggcctcag agacaagcca ctgaagttta agaaggcgaa gaaagacaag
      721 aaggaggaca aagaaggcaa gcatgagcca ctacaacctt cagcccacca ttctgcagag
      781 ccagcagagg caggcaaagc agaaacatca gaaagctcag gctctgcccc agcagtgcca
      841 gaagcctcgg cttcccccaa acagcggcgc tccattatcc gtgaccgggg acctatgtat
      901 gatgacccca ccttgcctga aggttggaca cgaaagctta aacaaaggaa gtctggccga
      961 tctgctggaa agtatgatgt atatttgatc aatccccagg gaaaagcttt tcgctctaaa
     1021 gtagaattga ttgcatactt tgaaaaggtg ggagacacct ccttggaccc taatgatttt
     1081 gacttcacgg taactgggag agggagcccc tccaggagag agcagaaacc acctaagaag
     1141 cccaaatctc ccaaagctcc aggaactggc aggggtcggg gacgccccaa agggagcggc
     1201 actgggagac caaaggcagc agcatcagaa ggtgttcagg tgaaaagggt cctggagaag
     1261 agccctggga aacttgttgt caagatgcct ttccaagcat cgcctggggg taagggtgag
     1321 ggaggtgggg ctaccacatc tgcccaggtc atggtgatca aacgccctgg cagaaagcga
     1381 aaagctgaag ctgaccccca ggccattcct aagaaacggg gtagaaagcc tgggagtgtg
     1441 gtggcagctg ctgcagctga ggccaaaaag aaagccgtga aggagtcttc catacggtct
     1501 gtgcatgaga ctgtgctccc catcaagaag cgcaagaccc gggagacggt cagcatcgag
     1561 gtcaaggaag tggtgaagcc cctgctggtg tccacccttg gtgagaaaag cgggaaggga
     1621 ctgaagacct gcaagagccc tgggcgtaaa agcaaggaga gcagccccaa ggggcgcagc
     1681 agcagtgcct cctccccacc taagaaggag caccatcatc accaccatca ctcagagtcc
     1741 acaaaggccc ccatgccact gctcccatcc ccacccccac ctgagcctga gagctctgag
     1801 gaccccatca gcccccctga gcctcaggac ttgagcagca gcatctgcaa agaagagaag
     1861 atgccccgag gaggctcact ggaaagcgat ggctgcccca aggagccagc taagactcag
     1921 cctatggtcg ccaccactac cacagttgca gaaaagtaca aacaccgagg ggagggagag
     1981 cgcaaagaca ttgtttcatc ttccatgcca aggccaaaca gagaggagcc tgtggacagc
     2041 cggacgcccg tgaccgagag agttagctgt aaggatccac cggtcgccac catggtgagc
     2101 aagggcgagg agctgttcac cggggtggtg cccatcctgg tcgagctgga cggcgacgta
     2161 aacggccaca agttcagcgt gtccggcgag ggcgagggcg atgccaccta cggcaagctg
     2221 accctgaagt tcatctgcac caccggcaag ctgcccgtgc cctggcccac cctcgtgacc
     2281 accctgacct acggcgtgca gtgcttcagc cgctaccccg accacatgaa gcagcacgac
     2341 ttcttcaagt ccgccatgcc cgaaggctac gtccaggagc gcaccatctt cttcaaggac
     2401 gacggcaact acaagacccg cgccgaggtg aagttcgagg gcgacaccct ggtgaaccgc
     2461 atcgagctga agggcatcga cttcaaggag gacggcaaca tcctggggca caagctggag
     2521 tacaactaca acagccacaa cgtctatatc atggccgaca agcagaagaa cggcatcaag
     2581 gtgaacttca agatccgcca caacatcgag gacggcagcg tgcagctcgc cgaccactac
     2641 cagcagaaca cccccatcgg cgacggcccc gtgctgctgc ccgacaacca ctacctgagc
     2701 acccagtccg ccctgagcaa agaccccaac gagaagcgcg atcacatggt cctgctggag
     2761 ttcgtgaccg ccgccgggat cactctcggc atggacgagc tgtacaagta aagcggccgc
     2821 gactctagat cataatcagc cataccacat ttgtagaggt tttacttgct ttaaaaaacc
     2881 tcccacacct ccccctgaac ctgaaacata aaatgaatgc aattgttgtt gttaacttgt
     2941 ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag
     3001 catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta tcttaaggcg
     3061 taaattgtaa gcgttaatat tttgttaaaa ttcgcgttaa atttttgtta aatcagctca
     3121 ttttttaacc aataggccga aatcggcaaa atcccttata aatcaaaaga atagaccgag
     3181 atagggttga gtgttgttcc agtttggaac aagagtccac tattaaagaa cgtggactcc
     3241 aacgtcaaag ggcgaaaaac cgtctatcag ggcgatggcc cactacgtga accatcaccc
     3301 taatcaagtt ttttggggtc gaggtgccgt aaagcactaa atcggaaccc taaagggagc
     3361 ccccgattta gagcttgacg gggaaagccg gcgaacgtgg cgagaaagga agggaagaaa
     3421 gcgaaaggag cgggcgctag ggcgctggca agtgtagcgg tcacgctgcg cgtaaccacc
     3481 acacccgccg cgcttaatgc gccgctacag ggcgcgtcag gtggcacttt tcggggaaat
     3541 gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg
     3601 agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtcc tgaggcggaa
     3661 agaaccagct gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc tccccagcag
     3721 gcagaagtat gcaaagcatg catctcaatt agtcagcaac caggtgtgga aagtccccag
     3781 gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accatagtcc
     3841 cgcccctaac tccgcccatc ccgcccctaa ctccgcccag ttccgcccat tctccgcccc
     3901 atggctgact aatttttttt atttatgcag aggccgaggc cgcctcggcc tctgagctat
     3961 tccagaagta gtgaggaggc ttttttggag gcctaggctt ttgcaaagat cgatcaagag
     4021 acaggatgag gatcgtttcg catgattgaa caagatggat tgcacgcagg ttctccggcc
     4081 gcttgggtgg agaggctatt cggctatgac tgggcacaac agacaatcgg ctgctctgat
     4141 gccgccgtgt tccggctgtc agcgcagggg cgcccggttc tttttgtcaa gaccgacctg
     4201 tccggtgccc tgaatgaact gcaagacgag gcagcgcggc tatcgtggct ggccacgacg
     4261 ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga ctggctgcta
     4321 ttgggcgaag tgccggggca ggatctcctg tcatctcacc ttgctcctgc cgagaaagta
     4381 tccatcatgg ctgatgcaat gcggcggctg catacgcttg atccggctac ctgcccattc
     4441 gaccaccaag cgaaacatcg catcgagcga gcacgtactc ggatggaagc cggtcttgtc
     4501 gatcaggatg atctggacga agagcatcag gggctcgcgc cagccgaact gttcgccagg
     4561 ctcaaggcga gcatgcccga cggcgaggat ctcgtcgtga cccatggcga tgcctgcttg
     4621 ccgaatatca tggtggaaaa tggccgcttt tctggattca tcgactgtgg ccggctgggt
     4681 gtggcggacc gctatcagga catagcgttg gctacccgtg atattgctga agagcttggc
     4741 ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc
     4801 atcgccttct atcgccttct tgacgagttc ttctgagcgg gactctgggg ttcgaaatga
     4861 ccgaccaagc gacgcccaac ctgccatcac gagatttcga ttccaccgcc gccttctatg
     4921 aaaggttggg cttcggaatc gttttccggg acgccggctg gatgatcctc cagcgcgggg
     4981 atctcatgct ggagttcttc gcccacccta gggggaggct aactgaaaca cggaaggaga
     5041 caataccgga aggaacccgc gctatgacgg caataaaaag acagaataaa acgcacggtg
     5101 ttgggtcgtt tgttcataaa cgcggggttc ggtcccaggg ctggcactct gtcgataccc
     5161 caccgagacc ccattggggc caatacgccc gcgtttcttc cttttcccca ccccaccccc
     5221 caagttcggg tgaaggccca gggctcgcag ccaacgtcgg ggcggcaggc cctgccatag
     5281 cctcaggtta ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga
     5341 tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt
     5401 tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc
     5461 tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc
     5521 cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac
     5581 caaatactgt tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac
     5641 cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt
     5701 cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct
     5761 gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat
     5821 acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt
     5881 atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg
     5941 cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt
     6001 gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt
     6061 tcctggcctt ttgctggcct tttgctcaca tgttctttcc tgcgttatcc cctgattctg
     6121 tggataaccg tattaccgcc atgcattag
//
