LOCUS       sequence-271780-        6617 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of MBD2a.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6617)
  AUTHORS   Wischnewski F, Friese O, Pantel K, Schwarzenbach H
  TITLE     Methyl-CpG binding domain proteins and their involvement in the 
            regulation of the MAGE-A1, MAGE-A2, MAGE-A3, and MAGE-A12 gene 
            promoters.
  JOURNAL   Mol Cancer Res. 2007 Jul;5(7):749-59.
  PUBMED    17634428
REFERENCE   2  (bases 1 to 6617)
  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: "Mol Cancer 
            Res."; date: "2007-07"; volume: "5(7)"; pages: "749-59"
COMMENT     Sequence Label: MBD2a pcDNA3.1
FEATURES             Location/Qualifiers
     source          1..6617
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..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"
     primer_bind     629..648
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        629..647
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1974..1991)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1980..2204
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2250..2678
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2337..2356)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2547..2568
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2687..2707)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2692..3021
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2872..3007
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2934..2953
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3088..3882
                     /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
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(3142..3161)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3752..3771
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4056..4177
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4093..4112)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4147..4166
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4226..4242)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4226..4242)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4239..4261)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4250..4266
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(4274..4280,4281..4298,4299..4304))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4319..4340
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4457..4474)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4628..5216)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4708..4727)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5387..6178,6179..6247))
                     /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     6010..6029
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6248..6352)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(6427..6446)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 gacattgatt attgactagt 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 agctctctgg ctaactagag
      601 aacccactgc ttactggctt atcgaaatta atacgactca ctatagggag acccaagctg
      661 gctagcgttt aaacttaagc ttggtaccga gctcggatcc atgcgcgcgc acccgggggg
      721 aggccgctgc tgcccggagc aggaggaggg ggagagtgcg gcgggcggca gcggcgctgg
      781 cggcgactcc gccatagagc aggggggcca gggcagcgcg ctcgccccgt ccccggtgag
      841 cggcgtgcgc agggaaggcg ctcggggcgg cggccgtggc cgggggcggt ggaagcaggc
      901 gggccggggc ggcggcgtct gtggccgtgg ccggggccgg ggccgtggcc ggggacgggg
      961 acggggccgg ggccggggcc gcggccgtcc cccgagtggc ggcagcggcc ttggcggcga
     1021 cggcggcggc tgcggcggcg gcggcagcgg tggcggcggc gccccccggc gggagccggt
     1081 ccctttcccg tcggggagcg cggggccggg gcccagggga ccccgggcca cggagagcgg
     1141 gaagaggatg gattgcccgg ccctcccccc cggatggaag aaggaggaag tgatccgaaa
     1201 atctgggcta agtgctggca agagcgatgt ctactacttc agtccaagtg gtaagaagtt
     1261 cagaagcaag cctcagttgg caaggtacct gggaaatact gttgatctca gcagttttga
     1321 cttcagaact ggaaagatga tgcctagtaa attacagaag aacaaacaga gactgcgaaa
     1381 cgatcctctc aatcaaaata agggtaaacc agacttgaat acaacattgc caattagaca
     1441 aacagcatca attttcaaac aaccggtaac caaagtcaca aatcatccta gtaataaagt
     1501 gaaatcagac ccacaacgaa tgaatgaaca gccacgtcag cttttctggg agaagaggct
     1561 acaaggactt agtgcatcag atgtaacaga acaaattata aaaaccatgg aactacccaa
     1621 aggtcttcaa ggagttggtc caggtagcaa tgatgagacc cttttatctg ctgttgccag
     1681 tgctttgcac acaagctctg cgccaatcac agggcaagtc tccgctgctg tggaaaagaa
     1741 ccctgctgtt tggcttaaca catctcaacc cctctgcaaa gcttttattg tcacagatga
     1801 agacatcagg aaacaggaag agcgagtaca gcaagtacgc aagaaattgg aagaagcact
     1861 gatggcagac atcttgtcgc gagctgctga tacagaagag atggatattg aaatggacag
     1921 tggagatgaa gcctaactcg agtctagagg gcccgtttaa acccgctgat cagcctcgac
     1981 tgtgccttct agttgccagc catctgttgt ttgcccctcc cccgtgcctt ccttgaccct
     2041 ggaaggtgcc actcccactg tcctttccta ataaaatgag gaaattgcat cgcattgtct
     2101 gagtaggtgt cattctattc tggggggtgg ggtggggcag gacagcaagg gggaggattg
     2161 ggaagacaat agcaggcatg ctggggatgc ggtgggctct atggcttctg aggcggaaag
     2221 aaccagctgg ggctctaggg ggtatcccca cgcgccctgt agcggcgcat taagcgcggc
     2281 gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc
     2341 tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa
     2401 tcgggggctc cctttagggt tccgatttag tgctttacgg cacctcgacc ccaaaaaact
     2461 tgattagggt gatggttcac gtagtgggcc atcgccctga tagacggttt ttcgcccttt
     2521 gacgttggag tccacgttct ttaatagtgg actcttgttc caaactggaa caacactcaa
     2581 ccctatctcg gtctattctt ttgatttata agggattttg ccgatttcgg cctattggtt
     2641 aaaaaatgag ctgatttaac aaaaatttaa cgcgaattaa ttctgtggaa tgtgtgtcag
     2701 ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc
     2761 aattagtcag caaccaggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa
     2821 agcatgcatc tcaattagtc agcaaccata gtcccgcccc taactccgcc catcccgccc
     2881 ctaactccgc ccagttccgc ccattctccg ccccatggct gactaatttt ttttatttat
     2941 gcagaggccg aggccgcctc tgcctctgag ctattccaga agtagtgagg aggctttttt
     3001 ggaggcctag gcttttgcaa aaagctcccg ggagcttgta tatccatttt cggatctgat
     3061 caagagacag gatgaggatc gtttcgcatg attgaacaag atggattgca cgcaggttct
     3121 ccggccgctt gggtggagag gctattcggc tatgactggg cacaacagac aatcggctgc
     3181 tctgatgccg ccgtgttccg gctgtcagcg caggggcgcc cggttctttt tgtcaagacc
     3241 gacctgtccg gtgccctgaa tgaactgcag gacgaggcag cgcggctatc gtggctggcc
     3301 acgacgggcg ttccttgcgc agctgtgctc gacgttgtca ctgaagcggg aagggactgg
     3361 ctgctattgg gcgaagtgcc ggggcaggat ctcctgtcat ctcaccttgc tcctgccgag
     3421 aaagtatcca tcatggctga tgcaatgcgg cggctgcata cgcttgatcc ggctacctgc
     3481 ccattcgacc accaagcgaa acatcgcatc gagcgagcac gtactcggat ggaagccggt
     3541 cttgtcgatc aggatgatct ggacgaagag catcaggggc tcgcgccagc cgaactgttc
     3601 gccaggctca aggcgcgcat gcccgacggc gaggatctcg tcgtgaccca tggcgatgcc
     3661 tgcttgccga atatcatggt ggaaaatggc cgcttttctg gattcatcga ctgtggccgg
     3721 ctgggtgtgg cggaccgcta tcaggacata gcgttggcta cccgtgatat tgctgaagag
     3781 cttggcggcg aatgggctga ccgcttcctc gtgctttacg gtatcgccgc tcccgattcg
     3841 cagcgcatcg ccttctatcg ccttcttgac gagttcttct gagcgggact ctggggttcg
     3901 aaatgaccga ccaagcgacg cccaacctgc catcacgaga tttcgattcc accgccgcct
     3961 tctatgaaag gttgggcttc ggaatcgttt tccgggacgc cggctggatg atcctccagc
     4021 gcggggatct catgctggag ttcttcgccc accccaactt gtttattgca gcttataatg
     4081 gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt
     4141 ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca tgtctgtata ccgtcgacct
     4201 ctagctagag cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc
     4261 tcacaattcc acacaacata cgagccggaa gcataaagtg taaagcctgg ggtgcctaat
     4321 gagtgagcta actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc
     4381 tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg
     4441 ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag
     4501 cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag
     4561 gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc
     4621 tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc
     4681 agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc
     4741 tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt
     4801 cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg
     4861 ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat
     4921 ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag
     4981 ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt
     5041 ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc
     5101 cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta
     5161 gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag
     5221 atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga
     5281 ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa
     5341 gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa
     5401 tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc
     5461 ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga
     5521 taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa
     5581 gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt
     5641 gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg
     5701 ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc
     5761 aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg
     5821 gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag
     5881 cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt
     5941 actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt
     6001 caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac
     6061 gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac
     6121 ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag
     6181 caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa
     6241 tactcatact cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga
     6301 gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc
     6361 cccgaaaagt gccacctgac gtcgacggat cgggagatct cccgatcccc tatggtgcac
     6421 tctcagtaca atctgctctg atgccgcata gttaagccag tatctgctcc ctgcttgtgt
     6481 gttggaggtc gctgagtagt gcgcgagcaa aatttaagct acaacaaggc aaggcttgac
     6541 cgacaattgc atgaagaatc tgcttagggt taggcgtttt gcgctgcttc gcgatgtacg
     6601 ggccagatat acgcgtt
//
