LOCUS       sequence-386850-        6245 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  The complete rat MeCp2 ORF is fused in frame of the NH2-terminus of 
            the enhanced GFP (isolated from pEGFP-N1 vector; CLONTECH 
            Laboratories, Inc.).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6245)
  AUTHORS   Brero A, Easwaran HP, Nowak D, Grunewald I, Cremer T, Leonhardt H, 
            Cardoso MC
  TITLE     Methyl CpG-binding proteins induce large-scale chromatin 
            reorganization during terminal differentiation.
  JOURNAL   J Cell Biol. 2005 Jun 6;169(5):733-43. doi: 10.1083/jcb.200502062.
  PUBMED    15939760
REFERENCE   2  (bases 1 to 6245)
  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: "10.1083/jcb"; 
            journalName: "J Cell Biol"; date: "2005-06-6- 6"; volume: "169"; 
            issue: "5"; pages: "733-43"
COMMENT     Sequence Label: pMeCP2G pc1121
FEATURES             Location/Qualifiers
     source          1..6245
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        78..381
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        382..585
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     536..556
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             1281..2138
                     /codon_start=1
                     /product="transcriptional repression domain from rat 
                     methyl-CpG-binding protein 2 (Nan et al., 1997)"
                     /label=MeCP2
                     /translation="VQVKRVLEKSPGKLLVKMPFQASPGGKGEGGGATTSAQVMVIKRP
                     GRKRKAEADPQAIPKKRGRKPGSVVAAAAAEAKKKAVKESSIRSVQETVLPIKKRKTRE
                     TVSIEVKEVVKPLLVSTLGEKSGKGLKTCKSPGRKSKESSPKGRSSSASSPPKKEHHHH
                     HHHAESPKAPMPLLPPPPPPEPQSSEDPISPPEPQDLSSSICKEEKMPRAGSLESDGCP
                     KEPAKTQPMVAAAATTTTTTTTTVAEKYKHRGEGERKDIVSSSMPRPNREEPVDSRTPV
                     TERVS"
     regulatory      2202..2211
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(2208..2210,2211..2213,2214..2927)
                     /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(2208..2210,2211..2213,2214..2924)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(2253..2274)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2514..2533)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2861..2882
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    3050..3171
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3087..3106)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3141..3160
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3178..3633)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3315..3336)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3527..3546
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3660..3764
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3766..4123
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3766..3957
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3789..3809)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3974..4109
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4158..4952
                     /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             4158..4952
                     /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(4212..4231)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4822..4841
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5140..5159)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5184..5231
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5560..6148
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6049..6068
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gtattaccgc catgcattag ttattaatag taatcaatta cggggtcatt agttcatagc
       61 ccatatatgg agttccgcgt tacataactt acggtaaatg gcccgcctgg ctgaccgccc
      121 aacgaccccc gcccattgac gtcaataatg acgtatgttc ccatagtaac gccaataggg
      181 actttccatt gacgtcaatg ggtggagtat ttacggtaaa ctgcccactt ggcagtacat
      241 caagtgtatc atatgccaag tacgccccct attgacgtca atgacggtaa atggcccgcc
      301 tggcattatg cccagtacat gaccttatgg gactttccta cttggcagta catctacgta
      361 ttagtcatcg ctattaccat ggtgatgcgg ttttggcagt acatcaatgg gcgtggatag
      421 cggtttgact cacggggatt tccaagtctc caccccattg acgtcaatgg gagtttgttt
      481 tggcaccaaa atcaacggga ctttccaaaa tgtcgtaaca actccgcccc attgacgcaa
      541 atgggcggta ggcgtgtacg gtgggaggtc tatataagca gagctggttt agtgaaccgt
      601 cagatccgct agcgctaccg gactcagatc tcgagctcca ccgcggtggc ggccgctcta
      661 gcatggtagc tgggatgtta gggctcagga aggaaaagtc agaagaccag gatctccagg
      721 gcctcaaaga gaaacccctg aagtttaaga aggtgaagaa agacaagaag gaagacaaag
      781 agggcaaaca tgaaccacta cagccttcag cccaccattc tgcagagcca gcagaggcag
      841 gcaaagcaga gacatcagaa agctcaggct ctgccccagc agtaccagaa gcctctgctt
      901 ctcccaaaca gcgacgttcc atcattcgtg accggggacc tatgtatgat gaccccacct
      961 tgcctgaagg ttggacgcga aagcttaaac agaggaagtc tggtcgctct gctggaaagt
     1021 atgatgtata tttgatcaat ccccagggaa aagcctttcg ctctaaagta gaattgattg
     1081 catattttga aaaggtggga gacacctcct tggaccctaa tgattttgac ttcactgtaa
     1141 ctgggagagg gagcccttcc aggagagaac agaaaccacc taagaagccc aaatctccca
     1201 aagctccagg aactggcagg ggtcggggac gcccgaaagg gagcggcact gggagaccaa
     1261 aggcagcagc atcagaaggt gttcaagtga aaagggtcct ggagaagagc cctgggaaac
     1321 ttctcgtcaa gatgcctttc caagcatcac ctgggggtaa gggtgaggga ggtggggcta
     1381 ccacatctgc gcaggtcatg gtgatcaaac gccctggcag aaagcgaaaa gctgaagctg
     1441 acccccaggc cattcctaag aaacggggta gaaagcctgg gagtgtggtg gcagctgctg
     1501 ctgcagaggc caaaaagaaa gctgtgaagg aatcttctat acggtctgtg caggagactg
     1561 tgctccccat caagaagcgc aagacccggg aaaccgtcag cattgaggtc aaggaggtgg
     1621 tgaagcccct gctggtgtct acacttggtg agaagagtgg aaagggactg aagacatgca
     1681 agagccctgg gcgtaaaagc aaggagagca gccccaaggg gcgcagcagc agtgcctcct
     1741 caccacctaa gaaggagcac catcatcacc accatcacgc agagtcccca aaggccccca
     1801 tgccattgct tccacctcca cccccacctg agcctcagag ctctgaggac cccatcagcc
     1861 cccctgagcc tcaggacttg agcagcagca tctgcaaaga ggagaagatg ccccgagcag
     1921 gctcactgga aagcgatggc tgccccaagg agccagctaa gactcagccc atggttgctg
     1981 ccgccgccac caccaccacc accaccacca ccacagttgc agaaaagtac aaacaccgag
     2041 gggagggaga gcgcaaagac attgtttcat cctccatgcc gaggccaaac agagaggagc
     2101 ctgtggacag ccggacgccc gtgaccgaga gagttagcag gaattcgata tcaagcttat
     2161 cgataccgtc gacggtaccg cgggcccggg atccaccggt cgccaccatg gtgagcaagg
     2221 gcgaggagct gttcaccggg gtggtgccca tcctggtcga gctggacggc gacgtaaacg
     2281 gccacaagtt cagcgtgtcc ggcgagggcg agggcgatgc cacctacggc aagctgaccc
     2341 tgaagttcat ctgcaccacc ggcaagctgc ccgtgccctg gcccaccctc gtgaccaccc
     2401 tgacctacgg cgtgcagtgc ttcagccgct accccgacca catgaagcag cacgacttct
     2461 tcaagtccgc catgcccgaa ggctacgtcc aggagcgcac catcttcttc aaggacgacg
     2521 gcaactacaa gacccgcgcc gaggtgaagt tcgagggcga caccctggtg aaccgcatcg
     2581 agctgaaggg catcgacttc aaggaggacg gcaacatcct ggggcacaag ctggagtaca
     2641 actacaacag ccacaacgtc tatatcatgg ccgacaagca gaagaacggc atcaaggtga
     2701 acttcaagat ccgccacaac atcgaggacg gcagcgtgca gctcgccgac cactaccagc
     2761 agaacacccc catcggcgac ggccccgtgc tgctgcccga caaccactac ctgagcaccc
     2821 agtccgccct gagcaaagac cccaacgaga agcgcgatca catggtcctg ctggagttcg
     2881 tgaccgccgc cgggatcact ctcggcatgg acgagctgta caagtaaagc ggccgcgact
     2941 ctagatcata atcagccata ccacatttgt agaggtttta cttgctttaa aaaacctccc
     3001 acacctcccc ctgaacctga aacataaaat gaatgcaatt gttgttgtta acttgtttat
     3061 tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt
     3121 tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt aaggcgtaaa
     3181 ttgtaagcgt taatattttg ttaaaattcg cgttaaattt ttgttaaatc agctcatttt
     3241 ttaaccaata ggccgaaatc ggcaaaatcc cttataaatc aaaagaatag accgagatag
     3301 ggttgagtgt tgttccagtt tggaacaaga gtccactatt aaagaacgtg gactccaacg
     3361 tcaaagggcg aaaaaccgtc tatcagggcg atggcccact acgtgaacca tcaccctaat
     3421 caagtttttt ggggtcgagg tgccgtaaag cactaaatcg gaaccctaaa gggagccccc
     3481 gatttagagc ttgacgggga aagccggcga acgtggcgag aaaggaaggg aagaaagcga
     3541 aaggagcggg cgctagggcg ctggcaagtg tagcggtcac gctgcgcgta accaccacac
     3601 ccgccgcgct taatgcgccg ctacagggcg cgtcaggtgg cacttttcgg ggaaatgtgc
     3661 gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac
     3721 aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtcctgag gcggaaagaa
     3781 ccagctgtgg aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc cagcaggcag
     3841 aagtatgcaa agcatgcatc tcaattagtc agcaaccagg tgtggaaagt ccccaggctc
     3901 cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca tagtcccgcc
     3961 cctaactccg cccatcccgc ccctaactcc gcccagttcc gcccattctc cgccccatgg
     4021 ctgactaatt ttttttattt atgcagaggc cgaggccgcc tcggcctctg agctattcca
     4081 gaagtagtga ggaggctttt ttggaggcct aggcttttgc aaagatcgat caagagacag
     4141 gatgaggatc gtttcgcatg attgaacaag atggattgca cgcaggttct ccggccgctt
     4201 gggtggagag gctattcggc tatgactggg cacaacagac aatcggctgc tctgatgccg
     4261 ccgtgttccg gctgtcagcg caggggcgcc cggttctttt tgtcaagacc gacctgtccg
     4321 gtgccctgaa tgaactgcaa gacgaggcag cgcggctatc gtggctggcc acgacgggcg
     4381 ttccttgcgc agctgtgctc gacgttgtca ctgaagcggg aagggactgg ctgctattgg
     4441 gcgaagtgcc ggggcaggat ctcctgtcat ctcaccttgc tcctgccgag aaagtatcca
     4501 tcatggctga tgcaatgcgg cggctgcata cgcttgatcc ggctacctgc ccattcgacc
     4561 accaagcgaa acatcgcatc gagcgagcac gtactcggat ggaagccggt cttgtcgatc
     4621 aggatgatct ggacgaagag catcaggggc tcgcgccagc cgaactgttc gccaggctca
     4681 aggcgagcat gcccgacggc gaggatctcg tcgtgaccca tggcgatgcc tgcttgccga
     4741 atatcatggt ggaaaatggc cgcttttctg gattcatcga ctgtggccgg ctgggtgtgg
     4801 cggaccgcta tcaggacata gcgttggcta cccgtgatat tgctgaagag cttggcggcg
     4861 aatgggctga ccgcttcctc gtgctttacg gtatcgccgc tcccgattcg cagcgcatcg
     4921 ccttctatcg ccttcttgac gagttcttct gagcgggact ctggggttcg aaatgaccga
     4981 ccaagcgacg cccaacctgc catcacgaga tttcgattcc accgccgcct tctatgaaag
     5041 gttgggcttc ggaatcgttt tccgggacgc cggctggatg atcctccagc gcggggatct
     5101 catgctggag ttcttcgccc accctagggg gaggctaact gaaacacgga aggagacaat
     5161 accggaagga acccgcgcta tgacggcaat aaaaagacag aataaaacgc acggtgttgg
     5221 gtcgtttgtt cataaacgcg gggttcggtc ccagggctgg cactctgtcg ataccccacc
     5281 gagaccccat tggggccaat acgcccgcgt ttcttccttt tccccacccc accccccaag
     5341 ttcgggtgaa ggcccagggc tcgcagccaa cgtcggggcg gcaggccctg ccatagcctc
     5401 aggttactca tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta
     5461 ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca
     5521 ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg
     5581 cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga
     5641 tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa
     5701 tactgttctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc
     5761 tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg
     5821 tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac
     5881 ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct
     5941 acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc
     6001 ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg
     6061 gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg
     6121 ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct
     6181 ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga
     6241 taacc
//
