LOCUS       sequence-386847-        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 YFP (pEYFP-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: pMeCP2Y pc644
FEATURES             Location/Qualifiers
     source          1..6245
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        77..380
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        381..584
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     535..555
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             1280..2137
                     /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      2201..2210
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(2207..2209,2210..2212,2213..2926)
                     /codon_start=1
                     /product="enhanced YFP"
                     /label=EYFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(2252..2273)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2513..2532)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2860..2881
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    3049..3170
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3086..3105)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3140..3159
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3177..3632)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3314..3335)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3526..3545
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3659..3763
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3765..4122
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3765..3956
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3788..3808)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3973..4108
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4157..4951
                     /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             4157..4951
                     /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(4211..4230)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4821..4840
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5139..5158)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5183..5230
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5559..6147
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6048..6067
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tattaccgcc atgcattagt tattaatagt aatcaattac ggggtcatta gttcatagcc
       61 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
      121 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
      181 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
      241 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
      301 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat
      361 tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc
      421 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt
      481 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa
      541 tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctggttta gtgaaccgtc
      601 agatccgcta gcgctaccgg actcagatct cgagctccac cgcggtggcg gccgctctag
      661 catggtagct gggatgttag ggctcaggaa ggaaaagtca gaagaccagg atctccaggg
      721 cctcaaagag aaacccctga agtttaagaa ggtgaagaaa gacaagaagg aagacaaaga
      781 gggcaaacat gaaccactac agccttcagc ccaccattct gcagagccag cagaggcagg
      841 caaagcagag acatcagaaa gctcaggctc tgccccagca gtaccagaag cctctgcttc
      901 tcccaaacag cgacgttcca tcattcgtga ccggggacct atgtatgatg accccacctt
      961 gcctgaaggt tggacgcgaa agcttaaaca gaggaagtct ggtcgctctg ctggaaagta
     1021 tgatgtatat ttgatcaatc cccagggaaa agcctttcgc tctaaagtag aattgattgc
     1081 atattttgaa aaggtgggag acacctcctt ggaccctaat gattttgact tcactgtaac
     1141 tgggagaggg agcccttcca ggagagaaca gaaaccacct aagaagccca aatctcccaa
     1201 agctccagga actggcaggg gtcggggacg cccgaaaggg agcggcactg ggagaccaaa
     1261 ggcagcagca tcagaaggtg ttcaagtgaa aagggtcctg gagaagagcc ctgggaaact
     1321 tctcgtcaag atgcctttcc aagcatcacc tgggggtaag ggtgagggag gtggggctac
     1381 cacatctgcg caggtcatgg tgatcaaacg ccctggcaga aagcgaaaag ctgaagctga
     1441 cccccaggcc attcctaaga aacggggtag aaagcctggg agtgtggtgg cagctgctgc
     1501 tgcagaggcc aaaaagaaag ctgtgaagga atcttctata cggtctgtgc aggagactgt
     1561 gctccccatc aagaagcgca agacccggga aaccgtcagc attgaggtca aggaggtggt
     1621 gaagcccctg ctggtgtcta cacttggtga gaagagtgga aagggactga agacatgcaa
     1681 gagccctggg cgtaaaagca aggagagcag ccccaagggg cgcagcagca gtgcctcctc
     1741 accacctaag aaggagcacc atcatcacca ccatcacgca gagtccccaa aggcccccat
     1801 gccattgctt ccacctccac ccccacctga gcctcagagc tctgaggacc ccatcagccc
     1861 ccctgagcct caggacttga gcagcagcat ctgcaaagag gagaagatgc cccgagcagg
     1921 ctcactggaa agcgatggct gccccaagga gccagctaag actcagccca tggttgctgc
     1981 cgccgccacc accaccacca ccaccaccac cacagttgca gaaaagtaca aacaccgagg
     2041 ggagggagag cgcaaagaca ttgtttcatc ctccatgccg aggccaaaca gagaggagcc
     2101 tgtggacagc cggacgcccg tgaccgagag agttagcagg aattcgatat caagcttatc
     2161 gataccgtcg acggtaccgc gggcccggga tccaccggtc gccaccatgg tgagcaaggg
     2221 cgaggagctg ttcaccgggg tggtgcccat cctggtcgag ctggacggcg acgtaaacgg
     2281 ccacaagttc agcgtgtccg gcgagggcga gggcgatgcc acctacggca agctgaccct
     2341 gaagttcatc tgcaccaccg gcaagctgcc cgtgccctgg cccaccctcg tgaccacctt
     2401 cggctacggc ctgcagtgct tcgcccgcta ccccgaccac atgaagcagc acgacttctt
     2461 caagtccgcc atgcccgaag gctacgtcca ggagcgcacc atcttcttca aggacgacgg
     2521 caactacaag acccgcgccg aggtgaagtt cgagggcgac accctggtga accgcatcga
     2581 gctgaagggc atcgacttca aggaggacgg caacatcctg gggcacaagc tggagtacaa
     2641 ctacaacagc cacaacgtct atatcatggc cgacaagcag aagaacggca tcaaggtgaa
     2701 cttcaagatc cgccacaaca tcgaggacgg cagcgtgcag ctcgccgacc actaccagca
     2761 gaacaccccc atcggcgacg gccccgtgct gctgcccgac aaccactacc tgagctacca
     2821 gtccgccctg agcaaagacc ccaacgagaa gcgcgatcac atggtcctgc tggagttcgt
     2881 gaccgccgcc gggatcactc tcggcatgga cgagctgtac aagtaaagcg gccgcgactc
     2941 tagatcataa tcagccatac cacatttgta gaggttttac ttgctttaaa aaacctccca
     3001 cacctccccc tgaacctgaa acataaaatg aatgcaattg ttgttgttaa cttgtttatt
     3061 gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt
     3121 ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta aggcgtaaat
     3181 tgtaagcgtt aatattttgt taaaattcgc gttaaatttt tgttaaatca gctcattttt
     3241 taaccaatag gccgaaatcg gcaaaatccc ttataaatca aaagaataga ccgagatagg
     3301 gttgagtgtt gttccagttt ggaacaagag tccactatta aagaacgtgg actccaacgt
     3361 caaagggcga aaaaccgtct atcagggcga tggcccacta cgtgaaccat caccctaatc
     3421 aagttttttg gggtcgaggt gccgtaaagc actaaatcgg aaccctaaag ggagcccccg
     3481 atttagagct tgacggggaa agccggcgaa cgtggcgaga aaggaaggga agaaagcgaa
     3541 aggagcgggc gctagggcgc tggcaagtgt agcggtcacg ctgcgcgtaa ccaccacacc
     3601 cgccgcgctt aatgcgccgc tacagggcgc gtcaggtggc acttttcggg gaaatgtgcg
     3661 cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca
     3721 ataaccctga taaatgcttc aataatattg aaaaaggaag agtcctgagg cggaaagaac
     3781 cagctgtgga atgtgtgtca gttagggtgt ggaaagtccc caggctcccc agcaggcaga
     3841 agtatgcaaa gcatgcatct caattagtca gcaaccaggt gtggaaagtc cccaggctcc
     3901 ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccat agtcccgccc
     3961 ctaactccgc ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc
     4021 tgactaattt tttttattta tgcagaggcc gaggccgcct cggcctctga gctattccag
     4081 aagtagtgag gaggcttttt tggaggccta ggcttttgca aagatcgatc aagagacagg
     4141 atgaggatcg tttcgcatga ttgaacaaga tggattgcac gcaggttctc cggccgcttg
     4201 ggtggagagg ctattcggct atgactgggc acaacagaca atcggctgct ctgatgccgc
     4261 cgtgttccgg ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg acctgtccgg
     4321 tgccctgaat gaactgcaag acgaggcagc gcggctatcg tggctggcca cgacgggcgt
     4381 tccttgcgca gctgtgctcg acgttgtcac tgaagcggga agggactggc tgctattggg
     4441 cgaagtgccg gggcaggatc tcctgtcatc tcaccttgct cctgccgaga aagtatccat
     4501 catggctgat gcaatgcggc ggctgcatac gcttgatccg gctacctgcc cattcgacca
     4561 ccaagcgaaa catcgcatcg agcgagcacg tactcggatg gaagccggtc ttgtcgatca
     4621 ggatgatctg gacgaagagc atcaggggct cgcgccagcc gaactgttcg ccaggctcaa
     4681 ggcgagcatg cccgacggcg aggatctcgt cgtgacccat ggcgatgcct gcttgccgaa
     4741 tatcatggtg gaaaatggcc gcttttctgg attcatcgac tgtggccggc tgggtgtggc
     4801 ggaccgctat caggacatag cgttggctac ccgtgatatt gctgaagagc ttggcggcga
     4861 atgggctgac cgcttcctcg tgctttacgg tatcgccgct cccgattcgc agcgcatcgc
     4921 cttctatcgc cttcttgacg agttcttctg agcgggactc tggggttcga aatgaccgac
     4981 caagcgacgc ccaacctgcc atcacgagat ttcgattcca ccgccgcctt ctatgaaagg
     5041 ttgggcttcg gaatcgtttt ccgggacgcc ggctggatga tcctccagcg cggggatctc
     5101 atgctggagt tcttcgccca ccctaggggg aggctaactg aaacacggaa ggagacaata
     5161 ccggaaggaa cccgcgctat gacggcaata aaaagacaga ataaaacgca cggtgttggg
     5221 tcgtttgttc ataaacgcgg ggttcggtcc cagggctggc actctgtcga taccccaccg
     5281 agaccccatt ggggccaata cgcccgcgtt tcttcctttt ccccacccca ccccccaagt
     5341 tcgggtgaag gcccagggct cgcagccaac gtcggggcgg caggccctgc catagcctca
     5401 ggttactcat atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag
     5461 gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac
     5521 tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc
     5581 gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat
     5641 caagagctac caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat
     5701 actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct
     5761 acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt
     5821 cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg
     5881 gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta
     5941 cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg
     6001 gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg
     6061 tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc
     6121 tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg
     6181 gccttttgct ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat
     6241 aaccg
//
