LOCUS       sequence-422492-        5962 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses fl. MBD2 GFP tagged in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5962)
  AUTHORS   Becker A, Allmann L, Hofstatter M, Casa V, Weber P, Lehmkuhl A, 
            Herce HD, Cardoso MC
  TITLE     Direct homo- and hetero-interactions of MeCP2 and MBD2.
  JOURNAL   PLoS One. 2013;8(1):e53730. doi: 10.1371/journal.pone.0053730. Epub 
            2013 Jan 15.
  PUBMED    23335972
REFERENCE   2  (bases 1 to 5962)
  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"; journalName: "PLoS One.";
            date: "2013"; volume: "8(1):e53730. doi"; pages: " 
            10.1371/journal.pone.0053730. Epub 2013 Jan 15"
COMMENT     Sequence Label: peMBD2G (pc2399)
FEATURES             Location/Qualifiers
     source          1..5962
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        80..383
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        384..587
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     538..558
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             join(1927..1929,1930..1932,1933..2646)
                     /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(1927..1929,1930..1932,1933..2643)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1972..1993)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2233..2252)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2580..2601
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    2769..2890
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2806..2825)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2860..2879
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2897..3352)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3034..3055)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3246..3265
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3379..3483
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3485..3842
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3485..3676
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3508..3528)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3693..3828
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3877..4671
                     /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             3877..4671
                     /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(3931..3950)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4541..4560
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4859..4878)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4903..4950
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5279..5867
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5768..5787
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ccgtattacc gccatgcatt agttattaat agtaatcaat tacggggtca ttagttcata
       61 gcccatatat ggagttccgc gttacataac ttacggtaaa tggcccgcct ggctgaccgc
      121 ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt tcccatagta acgccaatag
      181 ggactttcca ttgacgtcaa tgggtggagt atttacggta aactgcccac ttggcagtac
      241 atcaagtgta tcatatgcca agtacgcccc ctattgacgt caatgacggt aaatggcccg
      301 cctggcatta tgcccagtac atgaccttat gggactttcc tacttggcag tacatctacg
      361 tattagtcat cgctattacc atggtgatgc ggttttggca gtacatcaat gggcgtggat
      421 agcggtttga ctcacgggga tttccaagtc tccaccccat tgacgtcaat gggagtttgt
      481 tttggcacca aaatcaacgg gactttccaa aatgtcgtaa caactccgcc ccattgacgc
      541 aaatgggcgg taggcgtgta cggtgggagg tctatataag cagagctggt ttagtgaacc
      601 gtcagatccg ctagcgctac cggactcaga tctcgagctc aagcttcgaa ttcaaaggcc
      661 tacgtcgacg ccatgcgcgc gcacccgggg ggaggccgct gctgcccgga gcaggaggag
      721 ggggagagcg cggcgggcgg cagcggcgct ggcggcgact ccgccataga gcaggggggc
      781 cagggcagcg cgctcgctcc gtccccggtg agcggcgtgc gcagggaagg cgctcggggc
      841 ggcggccgtg gccgggggcg gtggaagcag gcggcccggg gcggcggcgt ctgtggccgt
      901 ggccgtggcc gtggccgggg tcggggccgt ggccggggcc ggggccgggg ccgcggccgt
      961 ccccagagtg gcggcagcgg ccttggcggc gacggcggcg gcggcgcggg cggctgcggc
     1021 gtcggcagcg gtggcggcgt cgccccccgg cgggatcctg tccctttccc gtcggggagc
     1081 tcggggccgg ggcccagggg accccgggcc acggagagcg ggaagaggat ggactgcccg
     1141 gccctccccc ccggatggaa gaaggaggaa gtgatccgaa aatcagggct cagtgctggc
     1201 aagagcgatg tctactactt cagtccaagt ggtaagaagt tcagaagtaa acctcagctg
     1261 gcaagatacc tgggaaatgc tgttgacctt agcagttttg acttcaggac cggcaagatg
     1321 atgcctagta aattacagaa gaacaagcag agactccgga atgaccccct caatcagaac
     1381 aagggtaaac cagacctgaa cacaacattg ccaattagac aaactgcatc aattttcaag
     1441 caaccagtaa ccaaattcac gaaccacccg agcaataagg tgaagtcaga cccccagcgg
     1501 atgaatgaac aaccacgtca gcttttctgg gagaagaggc tacaaggact tagcgcatca
     1561 gatgtaacag aacaaattat aaaaaccatg gagctaccta aaggtcttca aggagtcggt
     1621 ccaggtagca atgacgagac ccttctgtct gctgtggcca gtgctttaca cacaagctct
     1681 gcgcccatca caggacaagt ctctgctgcc gtggaaaaga accctgctgt ttggcttaac
     1741 acatctcaac ccctctgcaa agctttcatt gttacagatg aagacattag gaaacaggaa
     1801 gagcgagtcc aacaagtacg caagaaactg gaggaggcac tgatggccga catcctgtcc
     1861 cgggctgcgg acacggagga agtagacatt gacatggacg atggagatga ggcgtgcggc
     1921 cgcgccatgg tgagcaaggg cgaggagctg ttcaccgggg tggtgcccat cctggtcgag
     1981 ctggacggcg acgtaaacgg ccacaagttc agcgtgtccg gcgagggcga gggcgatgcc
     2041 acctacggca agctgaccct gaagttcatc tgcaccaccg gcaagctgcc cgtgccctgg
     2101 cccaccctcg tgaccaccct gacctacggc gtgcagtgct tcagccgcta ccccgaccac
     2161 atgaagcagc acgacttctt caagtccgcc atgcccgaag gctacgtcca ggagcgcacc
     2221 atcttcttca aggacgacgg caactacaag acccgcgccg aggtgaagtt cgagggcgac
     2281 accctggtga accgcatcga gctgaagggc atcgacttca aggaggacgg caacatcctg
     2341 gggcacaagc tggagtacaa ctacaacagc cacaacgtct atatcatggc cgacaagcag
     2401 aagaacggca tcaaggtgaa cttcaagatc cgccacaaca tcgaggacgg cagcgtgcag
     2461 ctcgccgacc actaccagca gaacaccccc atcggcgacg gccccgtgct gctgcccgac
     2521 aaccactacc tgagcaccca gtccgccctg agcaaagacc ccaacgagaa gcgcgatcac
     2581 atggtcctgc tggagttcgt gaccgccgcc gggatcactc tcggcatgga cgagctgtac
     2641 aagtaaagcg gccgcgactc tagatcataa tcagccatac cacatttgta gaggttttac
     2701 ttgctttaaa aaacctccca cacctccccc tgaacctgaa acataaaatg aatgcaattg
     2761 ttgttgttaa cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa
     2821 atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca
     2881 atgtatctta aggcgtaaat tgtaagcgtt aatattttgt taaaattcgc gttaaatttt
     2941 tgttaaatca gctcattttt taaccaatag gccgaaatcg gcaaaatccc ttataaatca
     3001 aaagaataga ccgagatagg gttgagtgtt gttccagttt ggaacaagag tccactatta
     3061 aagaacgtgg actccaacgt caaagggcga aaaaccgtct atcagggcga tggcccacta
     3121 cgtgaaccat caccctaatc aagttttttg gggtcgaggt gccgtaaagc actaaatcgg
     3181 aaccctaaag ggagcccccg atttagagct tgacggggaa agccggcgaa cgtggcgaga
     3241 aaggaaggga agaaagcgaa aggagcgggc gctagggcgc tggcaagtgt agcggtcacg
     3301 ctgcgcgtaa ccaccacacc cgccgcgctt aatgcgccgc tacagggcgc gtcaggtggc
     3361 acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat
     3421 atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag
     3481 agtcctgagg cggaaagaac cagctgtgga atgtgtgtca gttagggtgt ggaaagtccc
     3541 caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccaggt
     3601 gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt
     3661 cagcaaccat agtcccgccc ctaactccgc ccatcccgcc cctaactccg cccagttccg
     3721 cccattctcc gccccatggc tgactaattt tttttattta tgcagaggcc gaggccgcct
     3781 cggcctctga gctattccag aagtagtgag gaggcttttt tggaggccta ggcttttgca
     3841 aagatcgatc aagagacagg atgaggatcg tttcgcatga ttgaacaaga tggattgcac
     3901 gcaggttctc cggccgcttg ggtggagagg ctattcggct atgactgggc acaacagaca
     3961 atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc ggttcttttt
     4021 gtcaagaccg acctgtccgg tgccctgaat gaactgcaag acgaggcagc gcggctatcg
     4081 tggctggcca cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac tgaagcggga
     4141 agggactggc tgctattggg cgaagtgccg gggcaggatc tcctgtcatc tcaccttgct
     4201 cctgccgaga aagtatccat catggctgat gcaatgcggc ggctgcatac gcttgatccg
     4261 gctacctgcc cattcgacca ccaagcgaaa catcgcatcg agcgagcacg tactcggatg
     4321 gaagccggtc ttgtcgatca ggatgatctg gacgaagagc atcaggggct cgcgccagcc
     4381 gaactgttcg ccaggctcaa ggcgagcatg cccgacggcg aggatctcgt cgtgacccat
     4441 ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg attcatcgac
     4501 tgtggccggc tgggtgtggc ggaccgctat caggacatag cgttggctac ccgtgatatt
     4561 gctgaagagc ttggcggcga atgggctgac cgcttcctcg tgctttacgg tatcgccgct
     4621 cccgattcgc agcgcatcgc cttctatcgc cttcttgacg agttcttctg agcgggactc
     4681 tggggttcga aatgaccgac caagcgacgc ccaacctgcc atcacgagat ttcgattcca
     4741 ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt ccgggacgcc ggctggatga
     4801 tcctccagcg cggggatctc atgctggagt tcttcgccca ccctaggggg aggctaactg
     4861 aaacacggaa ggagacaata ccggaaggaa cccgcgctat gacggcaata aaaagacaga
     4921 ataaaacgca cggtgttggg tcgtttgttc ataaacgcgg ggttcggtcc cagggctggc
     4981 actctgtcga taccccaccg agaccccatt ggggccaata cgcccgcgtt tcttcctttt
     5041 ccccacccca ccccccaagt tcgggtgaag gcccagggct cgcagccaac gtcggggcgg
     5101 caggccctgc catagcctca ggttactcat atatacttta gattgattta aaacttcatt
     5161 tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt
     5221 aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt
     5281 gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag
     5341 cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta actggcttca
     5401 gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca
     5461 agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg
     5521 ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg
     5581 cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct
     5641 acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga
     5701 gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc
     5761 ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg
     5821 agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg
     5881 cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc tttcctgcgt
     5941 tatcccctga ttctgtggat aa
//
