LOCUS       sequence-359689-        6208 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Fluorescently tagged MECP2, RFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6208)
  AUTHORS   Erdel F, Rademacher A, Vlijm R, Tunnermann J, Frank L, Weinmann R, 
            Schweigert E, Yserentant K, Hummert J, Bauer C, Schumacher S, Al 
            Alwash A, Normand C, Herten DP, Engelhardt J, Rippe K
  TITLE     Mouse Heterochromatin Adopts Digital Compaction States without 
            Showing Hallmarks of HP1-Driven Liquid-Liquid Phase Separation.
  JOURNAL   Mol Cell. 2020 Apr 16;78(2):236-249.e7. doi: 
            10.1016/j.molcel.2020.02.005. Epub 2020 Feb 25.
  PUBMED    32101700
REFERENCE   2  (bases 1 to 6208)
  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: 
            "10.1016/j.molcel.2020.02.005"; journalName: "Mol Cell"; date: 
            "2020-04-16- 16"; volume: "78"; issue: "2"; pages: "236-249.e7"
COMMENT     Sequence Label: pMECP2-RFP
FEATURES             Location/Qualifiers
     source          1..6208
                     /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"
     CDS             1783..1803
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=7xHis
                     /translation="HHHHHHH"
     regulatory      2155..2164
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2161..2874
                     /codon_start=1
                     /product="monomeric derivative of red fluorescent protein 
                     from Entacmaea quadricolor (Merzlyak et al., 2007)"
                     /label=TagRFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELIKENMHMKLYMEGTVNNHHFKCTSEGEGKPYEGTQTM
                     RIKVVEGGPLPFAFDILATSFMYGSRTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGG
                     VLTATQDTSLQDGCLIYNVKIRGVNFPSNGPVMQKKTLGWEANTEMLYPADGGLEGRSD
                     MALKLVGGGHLICNFKTTYRSKKPAKNLKMPGVYYVDHRLERIKEADKETYVEQHEVAV
                     ARYCDLPSKLGHKLN"
     polyA_signal    2996..3117
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3033..3052)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3087..3106
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3124..3579)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3261..3282)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3473..3492
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3606..3710
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3712..4069
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3712..3903
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3735..3755)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3920..4055
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4104..4898
                     /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             4104..4898
                     /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(4158..4177)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4768..4787
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5086..5105)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5130..5177
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5506..6094
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5995..6014
                     /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 caagcttcga attcctatgg ccgccgctgc cgccaccgcc
      661 gccgccgccg ccgcgccgag cggaggagga ggaggaggcg aggaggagag actggaggaa
      721 aagtcagaag accaggatct ccagggcctc agagacaagc cactgaagtt taagaaggcg
      781 aagaaagaca agaaggagga caaagaaggc aagcatgagc cactacaacc ttcagcccac
      841 cattctgcag agccagcaga ggcaggcaaa gcagaaacat cagaaagctc aggctctgcc
      901 ccagcagtgc cagaagcctc ggcttccccc aaacagcggc gctccattat ccgtgaccgg
      961 ggacctatgt atgatgaccc caccttgcct gaaggttgga cacgaaagct taaacaaagg
     1021 aagtctggcc gatctgctgg aaagtatgat gtatatttga tcaatcccca gggaaaagct
     1081 tttcgctcta aagtagaatt gattgcatac tttgaaaagg tgggagacac ctccttggac
     1141 cctaatgatt ttgacttcac ggtaactggg agagggagcc cctccaggag agagcagaaa
     1201 ccacctaaga agcccaaatc tcccaaagct ccaggaactg gcaggggtcg gggacgcccc
     1261 aaagggagcg gcactgggag accaaaggca gcagcatcag aaggtgttca ggtgaaaagg
     1321 gtcctggaga agagccctgg gaaacttgtt gtcaagatgc ctttccaagc atcgcctggg
     1381 ggtaagggtg agggaggtgg ggctaccaca tctgcccagg tcatggtgat caaacgccct
     1441 ggcagaaagc gaaaagctga agctgacccc caggccattc ctaagaaacg gggtagaaag
     1501 cctgggagtg tggtggcagc tgctgcagct gaggccaaaa agaaagccgt gaaggagtct
     1561 tccatacggt ctgtgcatga gactgtgctc cccatcaaga agcgcaagac ccgggagacg
     1621 gtcagcatcg aggtcaagga agtggtgaag cccctgctgg tgtccaccct tggtgagaaa
     1681 agcgggaagg gactgaagac ctgcaagagc cctgggcgta aaagcaagga gagcagcccc
     1741 aaggggcgca gcagcagtgc ctcctcccca cctaagaagg agcaccatca tcaccaccat
     1801 cactcagagt ccacaaaggc ccccatgcca ctgctcccat ccccaccccc acctgagcct
     1861 gagagctctg aggaccccat cagcccccct gagcctcagg acttgagcag cagcatctgc
     1921 aaagaagaga agatgccccg aggaggctca ctggaaagcg atggctgccc caaggagcca
     1981 gctaagactc agcctatggt cgccaccact accacagttg cagaaaagta caaacaccga
     2041 ggggagggag agcgcaaaga cattgtttca tcttccatgc caaggccaaa cagagaggag
     2101 cctgtggaca gccggacgcc cgtgaccgag agagttagcg gggatccacc ggtcgccacc
     2161 atggtgtcta agggcgaaga gctgattaag gagaacatgc acatgaagct gtacatggag
     2221 ggcaccgtga acaaccacca cttcaagtgc acatccgagg gcgaaggcaa gccctacgag
     2281 ggcacccaga ccatgagaat caaggtggtc gagggcggcc ctctcccctt cgccttcgac
     2341 atcctggcta ccagcttcat gtacggcagc agaaccttca tcaaccacac ccagggcatc
     2401 cccgacttct ttaagcagtc cttccctgag ggcttcacat gggagagagt caccacatac
     2461 gaagacgggg gcgtgctgac cgctacccag gacaccagcc tccaggacgg ctgcctcatc
     2521 tacaacgtca agatcagagg ggtgaacttc ccatccaacg gccctgtgat gcagaagaaa
     2581 acactcggct gggaggccaa caccgagatg ctgtaccccg ctgacggcgg cctggaaggc
     2641 agaagcgaca tggccctgaa gctcgtgggc gggggccacc tgatctgcaa cttcaagacc
     2701 acatacagat ccaagaaacc cgctaagaac ctcaagatgc ccggcgtcta ctatgtggac
     2761 cacagactgg aaagaatcaa ggaggccgac aaagagacct acgtcgagca gcacgaggtg
     2821 gctgtggcca gatactgcga cctccctagc aaactggggc acaaacttaa ttgagcggcc
     2881 gcgactctag atcataatca gccataccac atttgtagag gttttacttg ctttaaaaaa
     2941 cctcccacac ctccccctga acctgaaaca taaaatgaat gcaattgttg ttgttaactt
     3001 gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa
     3061 agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttaagg
     3121 cgtaaattgt aagcgttaat attttgttaa aattcgcgtt aaatttttgt taaatcagct
     3181 cattttttaa ccaataggcc gaaatcggca aaatccctta taaatcaaaa gaatagaccg
     3241 agatagggtt gagtgttgtt ccagtttgga acaagagtcc actattaaag aacgtggact
     3301 ccaacgtcaa agggcgaaaa accgtctatc agggcgatgg cccactacgt gaaccatcac
     3361 cctaatcaag ttttttgggg tcgaggtgcc gtaaagcact aaatcggaac cctaaaggga
     3421 gcccccgatt tagagcttga cggggaaagc cggcgaacgt ggcgagaaag gaagggaaga
     3481 aagcgaaagg agcgggcgct agggcgctgg caagtgtagc ggtcacgctg cgcgtaacca
     3541 ccacacccgc cgcgcttaat gcgccgctac agggcgcgtc aggtggcact tttcggggaa
     3601 atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca
     3661 tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt cctgaggcgg
     3721 aaagaaccag ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag gctccccagc
     3781 aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc
     3841 aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccatagt
     3901 cccgccccta actccgccca tcccgcccct aactccgccc agttccgccc attctccgcc
     3961 ccatggctga ctaatttttt ttatttatgc agaggccgag gccgcctcgg cctctgagct
     4021 attccagaag tagtgaggag gcttttttgg aggcctaggc ttttgcaaag atcgatcaag
     4081 agacaggatg aggatcgttt cgcatgattg aacaagatgg attgcacgca ggttctccgg
     4141 ccgcttgggt ggagaggcta ttcggctatg actgggcaca acagacaatc ggctgctctg
     4201 atgccgccgt gttccggctg tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc
     4261 tgtccggtgc cctgaatgaa ctgcaagacg aggcagcgcg gctatcgtgg ctggccacga
     4321 cgggcgttcc ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg gactggctgc
     4381 tattgggcga agtgccgggg caggatctcc tgtcatctca ccttgctcct gccgagaaag
     4441 tatccatcat ggctgatgca atgcggcggc tgcatacgct tgatccggct acctgcccat
     4501 tcgaccacca agcgaaacat cgcatcgagc gagcacgtac tcggatggaa gccggtcttg
     4561 tcgatcagga tgatctggac gaagagcatc aggggctcgc gccagccgaa ctgttcgcca
     4621 ggctcaaggc gagcatgccc gacggcgagg atctcgtcgt gacccatggc gatgcctgct
     4681 tgccgaatat catggtggaa aatggccgct tttctggatt catcgactgt ggccggctgg
     4741 gtgtggcgga ccgctatcag gacatagcgt tggctacccg tgatattgct gaagagcttg
     4801 gcggcgaatg ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc gattcgcagc
     4861 gcatcgcctt ctatcgcctt cttgacgagt tcttctgagc gggactctgg ggttcgaaat
     4921 gaccgaccaa gcgacgccca acctgccatc acgagatttc gattccaccg ccgccttcta
     4981 tgaaaggttg ggcttcggaa tcgttttccg ggacgccggc tggatgatcc tccagcgcgg
     5041 ggatctcatg ctggagttct tcgcccaccc tagggggagg ctaactgaaa cacggaagga
     5101 gacaataccg gaaggaaccc gcgctatgac ggcaataaaa agacagaata aaacgcacgg
     5161 tgttgggtcg tttgttcata aacgcggggt tcggtcccag ggctggcact ctgtcgatac
     5221 cccaccgaga ccccattggg gccaatacgc ccgcgtttct tccttttccc caccccaccc
     5281 cccaagttcg ggtgaaggcc cagggctcgc agccaacgtc ggggcggcag gccctgccat
     5341 agcctcaggt tactcatata tactttagat tgatttaaaa cttcattttt aatttaaaag
     5401 gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc
     5461 gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt
     5521 tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt
     5581 gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat
     5641 accaaatact gttcttctag tgtagccgta gttaggccac cacttcaaga actctgtagc
     5701 accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa
     5761 gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg
     5821 ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag
     5881 atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag
     5941 gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa
     6001 cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt
     6061 gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg
     6121 gttcctggcc ttttgctggc cttttgctca catgttcttt cctgcgttat cccctgattc
     6181 tgtggataac cgtattaccg ccatgcat
//
