LOCUS       sequence-207358-        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        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             join(1457..1459,1460..1462,1463..2176)
                     /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(1457..1459,1460..1462,1463..2173)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1502..1523)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1763..1782)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2110..2131
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    2299..2420
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2336..2355)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2390..2409
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2427..2882)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2564..2585)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2776..2795
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2909..3013
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3015..3372
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3015..3206
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3038..3058)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3223..3358
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3407..4201
                     /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             3407..4201
                     /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(3461..3480)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4071..4090
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4389..4408)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4433..4480
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4809..5397
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5298..5317
                     /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 cggtcgggag
     1441 ctccggcagt tctggaatgg tgagcaaggg cgaggagctg ttcaccgggg tggtgcccat
     1501 cctggtcgag ctggacggcg acgtaaacgg ccacaagttc agcgtgtccg gcgagggcga
     1561 gggcgatgcc acctacggca agctgaccct gaagttcatc tgcaccaccg gcaagctgcc
     1621 cgtgccctgg cccaccctcg tgaccaccct gacctacggc gtgcagtgct tcagccgcta
     1681 ccccgaccac atgaagcagc acgacttctt caagtccgcc atgcccgaag gctacgtcca
     1741 ggagcgcacc atcttcttca aggacgacgg caactacaag acccgcgccg aggtgaagtt
     1801 cgagggcgac accctggtga accgcatcga gctgaagggc atcgacttca aggaggacgg
     1861 caacatcctg gggcacaagc tggagtacaa ctacaacagc cacaacgtct atatcatggc
     1921 cgacaagcag aagaacggca tcaaggtgaa cttcaagatc cgccacaaca tcgaggacgg
     1981 cagcgtgcag ctcgccgacc actaccagca gaacaccccc atcggcgacg gccccgtgct
     2041 gctgcccgac aaccactacc tgagcaccca gtccgccctg agcaaagacc ccaacgagaa
     2101 gcgcgatcac atggtcctgc tggagttcgt gaccgccgcc gggatcactc tcggcatgga
     2161 cgagctgtac aagtaaagcg gccgcgactc tagatcataa tcagccatac cacatttgta
     2221 gaggttttac ttgctttaaa aaacctccca cacctccccc tgaacctgaa acataaaatg
     2281 aatgcaattg ttgttgttaa cttgtttatt gcagcttata atggttacaa ataaagcaat
     2341 agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc
     2401 aaactcatca atgtatctta aggcgtaaat tgtaagcgtt aatattttgt taaaattcgc
     2461 gttaaatttt tgttaaatca gctcattttt taaccaatag gccgaaatcg gcaaaatccc
     2521 ttataaatca aaagaataga ccgagatagg gttgagtgtt gttccagttt ggaacaagag
     2581 tccactatta aagaacgtgg actccaacgt caaagggcga aaaaccgtct atcagggcga
     2641 tggcccacta cgtgaaccat caccctaatc aagttttttg gggtcgaggt gccgtaaagc
     2701 actaaatcgg aaccctaaag ggagcccccg atttagagct tgacggggaa agccggcgaa
     2761 cgtggcgaga aaggaaggga agaaagcgaa aggagcgggc gctagggcgc tggcaagtgt
     2821 agcggtcacg ctgcgcgtaa ccaccacacc cgccgcgctt aatgcgccgc tacagggcgc
     2881 gtcaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat
     2941 acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg
     3001 aaaaaggaag agtcctgagg cggaaagaac cagctgtgga atgtgtgtca gttagggtgt
     3061 ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca
     3121 gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat
     3181 ctcaattagt cagcaaccat agtcccgccc ctaactccgc ccatcccgcc cctaactccg
     3241 cccagttccg cccattctcc gccccatggc tgactaattt tttttattta tgcagaggcc
     3301 gaggccgcct cggcctctga gctattccag aagtagtgag gaggcttttt tggaggccta
     3361 ggcttttgca aagatcgatc aagagacagg atgaggatcg tttcgcatga ttgaacaaga
     3421 tggattgcac gcaggttctc cggccgcttg ggtggagagg ctattcggct atgactgggc
     3481 acaacagaca atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc
     3541 ggttcttttt gtcaagaccg acctgtccgg tgccctgaat gaactgcaag acgaggcagc
     3601 gcggctatcg tggctggcca cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac
     3661 tgaagcggga agggactggc tgctattggg cgaagtgccg gggcaggatc tcctgtcatc
     3721 tcaccttgct cctgccgaga aagtatccat catggctgat gcaatgcggc ggctgcatac
     3781 gcttgatccg gctacctgcc cattcgacca ccaagcgaaa catcgcatcg agcgagcacg
     3841 tactcggatg gaagccggtc ttgtcgatca ggatgatctg gacgaagagc atcaggggct
     3901 cgcgccagcc gaactgttcg ccaggctcaa ggcgagcatg cccgacggcg aggatctcgt
     3961 cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg
     4021 attcatcgac tgtggccggc tgggtgtggc ggaccgctat caggacatag cgttggctac
     4081 ccgtgatatt gctgaagagc ttggcggcga atgggctgac cgcttcctcg tgctttacgg
     4141 tatcgccgct cccgattcgc agcgcatcgc cttctatcgc cttcttgacg agttcttctg
     4201 agcgggactc tggggttcga aatgaccgac caagcgacgc ccaacctgcc atcacgagat
     4261 ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt ccgggacgcc
     4321 ggctggatga tcctccagcg cggggatctc atgctggagt tcttcgccca ccctaggggg
     4381 aggctaactg aaacacggaa ggagacaata ccggaaggaa cccgcgctat gacggcaata
     4441 aaaagacaga ataaaacgca cggtgttggg tcgtttgttc ataaacgcgg ggttcggtcc
     4501 cagggctggc actctgtcga taccccaccg agaccccatt ggggccaata cgcccgcgtt
     4561 tcttcctttt ccccacccca ccccccaagt tcgggtgaag gcccagggct cgcagccaac
     4621 gtcggggcgg caggccctgc catagcctca ggttactcat atatacttta gattgattta
     4681 aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc
     4741 aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa
     4801 ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca
     4861 ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta
     4921 actggcttca gcagagcgca gataccaaat actgtccttc tagtgtagcc gtagttaggc
     4981 caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca
     5041 gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta
     5101 ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag
     5161 cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt
     5221 cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc
     5281 acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac
     5341 ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac
     5401 gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc
     5461 tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgccatgca t
//
