LOCUS       sequence-422744-        6250 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses MBD2 (aa153-414) GFP tagged in insect cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6250)
  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 6250)
  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: pFB_MBD2.2-GFP (pc2083)
FEATURES             Location/Qualifiers
     source          1..6250
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(84..103)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     293..314
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        477..581
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(582..650,651..1442)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(800..819)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1613..2201
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2102..2121
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2355..2372
                     /label=L4440
                     /note="L4440 vector, forward primer"
     mobile_element  2506..2730
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7R
                     /note="mini-Tn7 element (right end of the Tn7 transposon)"
     CDS             complement(2797..3330)
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLWSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     promoter        complement(join(3500..3505,3506..3522,3523..3528))
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     promoter        3880..3971
                     /gene="polh from Autographa californica"
                     /label=polyhedrin promoter
                     /note="promoter for the baculovirus polyhedrin gene"
     primer_bind     3896..3913
                     /label=Polyhedrin-F
                     /note="Polyhedrin promoter, forward primer"
     CDS             join(4853..4855,4856..4858,4859..5572)
                     /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(4853..4855,4856..4858,4859..5569)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(4898..4919)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(5159..5178)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     5506..5527
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    5735..5869
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5772..5791)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5826..5845
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     mobile_element  5898..6063
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7L
                     /note="mini-Tn7 element (left end of the Tn7 transposon)"
     rep_origin      join(6247..6250,1..451)
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
ORIGIN
        1 gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac
       61 acttgccagc gccctagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt
      121 cgccggcttt ccccgtcaag ctctaaatcg ggggctccct ttagggttcc gatttagtgc
      181 tttacggcac ctcgacccca aaaaacttga ttagggtgat ggttcacgta gtgggccatc
      241 gccctgatag acggtttttc gccctttgac gttggagtcc acgttcttaa tagtggactc
      301 ttgttccaaa ctggaacaac actcaaccct atctcggtct attcttttga tttataaggg
      361 attttgccga tttcggccta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg
      421 aattttaaca aaatattaac gcttacaatt taggtggcac ttttcgggga aatgtgcgcg
      481 gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat
      541 aaccctgata aatgcttcaa taatattgaa aaaggaagag tatgagtatt caacatttcc
      601 gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct cacccagaaa
      661 cgctggtgaa agtaaaagat gctgaagatc agttgggtgc acgagtgggt tacatcgaac
      721 tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgt tttccaatga
      781 tgagcacttt taaagttctg ctatgtggcg cggtattatc ccgtattgac gccgggcaag
      841 agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtac tcaccagtca
      901 cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct gccataacca
      961 tgagtgataa cactgcggcc aacttacttc tgacaacgat cggaggaccg aaggagctaa
     1021 ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg gaaccggagc
     1081 tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgtagca atggcaacaa
     1141 cgttgcgcaa actattaact ggcgaactac ttactctagc ttcccggcaa caattaatag
     1201 actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt ccggctggct
     1261 ggtttattgc tgataaatct ggagccggtg agcgtgggtc tcgcggtatc attgcagcac
     1321 tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg agtcaggcaa
     1381 ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt aagcattggt
     1441 aactgtcaga ccaagtttac tcatatatac tttagattga tttaaaactt catttttaat
     1501 ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg
     1561 agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc
     1621 ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg
     1681 tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag
     1741 cgcagatacc aaatactgtt cttctagtgt agccgtagtt aggccaccac ttcaagaact
     1801 ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg
     1861 gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc
     1921 ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg
     1981 aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg
     2041 cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag
     2101 ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc
     2161 gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct
     2221 ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct gcgttatccc
     2281 ctgattctgt ggataaccgt attaccgcct ttgagtgagc tgataccgct cgccgcagcc
     2341 gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga agagcgcctg atgcggtatt
     2401 ttctccttac gcatctgtgc ggtatttcac accgcataga ccagccgcgt aacctggcaa
     2461 aatcggttac ggttgagtaa taaatggatg ccctgcgtaa gcgggtgtgg gcggacaata
     2521 aagtcttaaa ctgaacaaaa tagatctaaa ctatgacaat aaagtcttaa actagacaga
     2581 atagttgtaa actgaaatca gtccagttat gctgtgaaaa agcatactgg acttttgtta
     2641 tggctaaagc aaactcttca ttttctgaag tgcaaattgc ccgtcgtatt aaagaggggc
     2701 gtggccaagg gcatggtaaa gactatattc gcggcgttgt gacaatttac cgaacaactc
     2761 cgcggccggg aagccgatct cggcttgaac gaattgttag gtggcggtac ttgggtcgat
     2821 atcaaagtgc atcacttctt cccgtatgcc caactttgta tagagagcca ctgcgggatc
     2881 gtcaccgtaa tctgcttgca cgtagatcac ataagcacca agcgcgttgg cctcatgctt
     2941 gaggagattg atgagcgcgg tggcaatgcc ctgcctccgg tgctcgccgg agactgcgag
     3001 atcatagata tagatctcac tacgcggctg ctcaaacttg ggcagaacgt aagccgcgag
     3061 agcgccaaca accgcttctt ggtcgaaggc agcaagcgcg atgaatgtct tactacggag
     3121 caagttcccg aggtaatcgg agtccggctg atgttgggag taggtggcta cgtctccgaa
     3181 ctcacgaccg aaaagatcaa gagcagcccg catggatttg acttggtcag ggccgagcct
     3241 acatgtgcga atgatgccca tacttgagcc acctaacttt gttttagggc gactgccctg
     3301 ctgcgtaaca tcgttgctgc tccataacat caaacatcga cccacggcgt aacgcgcttg
     3361 ctgcttggat gcccgaggca tagactgtac aaaaaaacag tcataacaag ccatgaaaac
     3421 cgccactgcg ccgttaccac cgctgcgttc ggtcaaggtt ctggaccagt tgcgtgagcg
     3481 catacgctac ttgcattaca gtttacgaac cgaacaggct tatgtcaact gggttcgtgc
     3541 cttcatccgt ttccacggtg tgcgtcaccc ggcaaccttg ggcagcagcg aagtcgaggc
     3601 atttctgtcc tggctggcga acgagcgcaa ggtttcggtc tccacgcatc gtcaggcatt
     3661 ggcggccttg ctgttcttct acggcaaggt gctgtgcacg gatctgccct ggcttcagga
     3721 gatcggaaga cctcggccgt cgcggcgctt gccggtggtg ctgaccccgg atgaagtggt
     3781 tcgcatcctc ggttttctgg aaggcgagca tcgtttgttc gcccaggact ctagctatag
     3841 ttctagtggt tggctacgta tactccggaa tattaataga tcatggagat aattaaaatg
     3901 ataaccatct cgcaaataaa taagtatttt actgttttcg taacagtttt gtaataaaaa
     3961 aacctataaa tattccggat tattcatacc gtcccaccat cgggcgcgga tcccggtccg
     4021 aagcgcgcgg aattcaaagg cctacgtcga cgccatggac tgcccggccc tcccccccgg
     4081 atggaagaag gaggaagtga tccgaaaatc agggctcagt gctggcaaga gcgatgtcta
     4141 ctacttcagt ccaagtggta agaagttcag aagtaaacct cagctggcaa gatacctggg
     4201 aaatgctgtt gaccttagca gttttgactt caggaccggc aagatgatgc ctagtaaatt
     4261 acagaagaac aagcagagac tccggaatga ccccctcaat cagaacaagg gtaaaccaga
     4321 cctgaacaca acattgccaa ttagacaaac tgcatcaatt ttcaagcaac cagtaaccaa
     4381 attcacgaac cacccgagca ataaggtgaa gtcagacccc cagcggatga atgaacaacc
     4441 acgtcagctt ttctgggaga agaggctaca aggacttagc gcatcagatg taacagaaca
     4501 aattataaaa accatggagc tacctaaagg tcttcaagga gtcggtccag gtagcaatga
     4561 cgagaccctt ctgtctgctg tggccagtgc tttacacaca agctctgcgc ccatcacagg
     4621 acaagtctct gctgccgtgg aaaagaaccc tgctgtttgg cttaacacat ctcaacccct
     4681 ctgcaaagct ttcattgtta cagatgaaga cattaggaaa caggaagagc gagtccaaca
     4741 agtacgcaag aaactggagg aggcactgat ggccgacatc ctgtcccggg ctgcggacac
     4801 ggaggaagta gacattgaca tggacgatgg agatgaggcg tgcggccgcg ccatggtgag
     4861 caagggcgag gagctgttca ccggggtggt gcccatcctg gtcgagctgg acggcgacgt
     4921 aaacggccac aagttcagcg tgtccggcga gggcgagggc gatgccacct acggcaagct
     4981 gaccctgaag ttcatctgca ccaccggcaa gctgcccgtg ccctggccca ccctcgtgac
     5041 caccctgacc tacggcgtgc agtgcttcag ccgctacccc gaccacatga agcagcacga
     5101 cttcttcaag tccgccatgc ccgaaggcta cgtccaggag cgcaccatct tcttcaagga
     5161 cgacggcaac tacaagaccc gcgccgaggt gaagttcgag ggcgacaccc tggtgaaccg
     5221 catcgagctg aagggcatcg acttcaagga ggacggcaac atcctggggc acaagctgga
     5281 gtacaactac aacagccaca acgtctatat catggccgac aagcagaaga acggcatcaa
     5341 ggtgaacttc aagatccgcc acaacatcga ggacggcagc gtgcagctcg ccgaccacta
     5401 ccagcagaac acccccatcg gcgacggccc cgtgctgctg cccgacaacc actacctgag
     5461 cacccagtcc gccctgagca aagaccccaa cgagaagcgc gatcacatgg tcctgctgga
     5521 gttcgtgacc gccgccggga tcactctcgg catggacgag ctgtacaagt aatctagagc
     5581 ctgcagtctc gaggcatgcg gtaccaagct tgtcgagaag tactagagga tcataatcag
     5641 ccataccaca tttgtagagg ttttacttgc tttaaaaaac ctcccacacc tccccctgaa
     5701 cctgaaacat aaaatgaatg caattgttgt tgttaacttg tttattgcag cttataatgg
     5761 ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc
     5821 tagttgtggt ttgtccaaac tcatcaatgt atcttatcat gtctggatct gatcactgct
     5881 tgagcctagg agatccgaac cagataagtg aaatctagtt ccaaactatt ttgtcatttt
     5941 taattttcgt attagcttac gacgctacac ccagttccca tctattttgt cactcttccc
     6001 taaataatcc ttaaaaactc catttccacc cctcccagtt cccaactatt ttgtccgccc
     6061 acagcggggc atttttcttc ctgttatgtt tttaatcaaa catcctgcca actccatgtg
     6121 acaaaccgtc atcttcggct actttttctc tgtcacagaa tgaaaatttt tctgtcatct
     6181 cttcgttatt aatgtttgta attgactgaa tatcaacgct tatttgcagc ctgaatggcg
     6241 aatgggacgc
//
