LOCUS       sequence-422501-        6706 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses fl. MBD2 GFP tagged in insect cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6706)
  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 6706)
  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 MBD2a-C-GFP (pc1565)
FEATURES             Location/Qualifiers
     source          1..6706
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(83..102)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     292..313
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        476..580
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(581..649,650..1441)
                     /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(799..818)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1612..2200
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2101..2120
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2354..2371
                     /label=L4440
                     /note="L4440 vector, forward primer"
     mobile_element  2505..2729
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7R
                     /note="mini-Tn7 element (right end of the Tn7 transposon)"
     CDS             complement(2796..3329)
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLWSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     promoter        complement(join(3499..3504,3505..3521,3522..3527))
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     promoter        3879..3970
                     /gene="polh from Autographa californica"
                     /label=polyhedrin promoter
                     /note="promoter for the baculovirus polyhedrin gene"
     primer_bind     3895..3912
                     /label=Polyhedrin-F
                     /note="Polyhedrin promoter, forward primer"
     CDS             join(5308..5310,5311..5313,5314..6027)
                     /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(5308..5310,5311..5313,5314..6024)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(5353..5374)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(5614..5633)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     5961..5982
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    6190..6324
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6227..6246)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6281..6300
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     mobile_element  6353..6518
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7L
                     /note="mini-Tn7 element (left end of the Tn7 transposon)"
     rep_origin      join(6702..6706,1..450)
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
ORIGIN
        1 ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca
       61 cttgccagcg ccctagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc
      121 gccggctttc cccgtcaagc tctaaatcgg gggctccctt tagggttccg atttagtgct
      181 ttacggcacc tcgaccccaa aaaacttgat tagggtgatg gttcacgtag tgggccatcg
      241 ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttcttaat agtggactct
      301 tgttccaaac tggaacaaca ctcaacccta tctcggtcta ttcttttgat ttataaggga
      361 ttttgccgat ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga
      421 attttaacaa aatattaacg cttacaattt aggtggcact tttcggggaa atgtgcgcgg
      481 aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata
      541 accctgataa atgcttcaat aatattgaaa aaggaagagt atgagtattc aacatttccg
      601 tgtcgccctt attccctttt ttgcggcatt ttgccttcct gtttttgctc acccagaaac
      661 gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt acatcgaact
      721 ggatctcaac agcggtaaga tccttgagag ttttcgcccc gaagaacgtt ttccaatgat
      781 gagcactttt aaagttctgc tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga
      841 gcaactcggt cgccgcatac actattctca gaatgacttg gttgagtact caccagtcac
      901 agaaaagcat cttacggatg gcatgacagt aagagaatta tgcagtgctg ccataaccat
      961 gagtgataac actgcggcca acttacttct gacaacgatc ggaggaccga aggagctaac
     1021 cgcttttttg cacaacatgg gggatcatgt aactcgcctt gatcgttggg aaccggagct
     1081 gaatgaagcc ataccaaacg acgagcgtga caccacgatg cctgtagcaa tggcaacaac
     1141 gttgcgcaaa ctattaactg gcgaactact tactctagct tcccggcaac aattaataga
     1201 ctggatggag gcggataaag ttgcaggacc acttctgcgc tcggcccttc cggctggctg
     1261 gtttattgct gataaatctg gagccggtga gcgtgggtct cgcggtatca ttgcagcact
     1321 ggggccagat ggtaagccct cccgtatcgt agttatctac acgacgggga gtcaggcaac
     1381 tatggatgaa cgaaatagac agatcgctga gataggtgcc tcactgatta agcattggta
     1441 actgtcagac caagtttact catatatact ttagattgat ttaaaacttc atttttaatt
     1501 taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga
     1561 gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc
     1621 tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt
     1681 ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc
     1741 gcagatacca aatactgttc ttctagtgta gccgtagtta ggccaccact tcaagaactc
     1801 tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg
     1861 cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg
     1921 gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga
     1981 actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc
     2041 ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg
     2101 gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg
     2161 atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt
     2221 tttacggttc ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc
     2281 tgattctgtg gataaccgta ttaccgcctt tgagtgagct gataccgctc gccgcagccg
     2341 aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa gagcgcctga tgcggtattt
     2401 tctccttacg catctgtgcg gtatttcaca ccgcatagac cagccgcgta acctggcaaa
     2461 atcggttacg gttgagtaat aaatggatgc cctgcgtaag cgggtgtggg cggacaataa
     2521 agtcttaaac tgaacaaaat agatctaaac tatgacaata aagtcttaaa ctagacagaa
     2581 tagttgtaaa ctgaaatcag tccagttatg ctgtgaaaaa gcatactgga cttttgttat
     2641 ggctaaagca aactcttcat tttctgaagt gcaaattgcc cgtcgtatta aagaggggcg
     2701 tggccaaggg catggtaaag actatattcg cggcgttgtg acaatttacc gaacaactcc
     2761 gcggccggga agccgatctc ggcttgaacg aattgttagg tggcggtact tgggtcgata
     2821 tcaaagtgca tcacttcttc ccgtatgccc aactttgtat agagagccac tgcgggatcg
     2881 tcaccgtaat ctgcttgcac gtagatcaca taagcaccaa gcgcgttggc ctcatgcttg
     2941 aggagattga tgagcgcggt ggcaatgccc tgcctccggt gctcgccgga gactgcgaga
     3001 tcatagatat agatctcact acgcggctgc tcaaacttgg gcagaacgta agccgcgaga
     3061 gcgccaacaa ccgcttcttg gtcgaaggca gcaagcgcga tgaatgtctt actacggagc
     3121 aagttcccga ggtaatcgga gtccggctga tgttgggagt aggtggctac gtctccgaac
     3181 tcacgaccga aaagatcaag agcagcccgc atggatttga cttggtcagg gccgagccta
     3241 catgtgcgaa tgatgcccat acttgagcca cctaactttg ttttagggcg actgccctgc
     3301 tgcgtaacat cgttgctgct ccataacatc aaacatcgac ccacggcgta acgcgcttgc
     3361 tgcttggatg cccgaggcat agactgtaca aaaaaacagt cataacaagc catgaaaacc
     3421 gccactgcgc cgttaccacc gctgcgttcg gtcaaggttc tggaccagtt gcgtgagcgc
     3481 atacgctact tgcattacag tttacgaacc gaacaggctt atgtcaactg ggttcgtgcc
     3541 ttcatccgtt tccacggtgt gcgtcacccg gcaaccttgg gcagcagcga agtcgaggca
     3601 tttctgtcct ggctggcgaa cgagcgcaag gtttcggtct ccacgcatcg tcaggcattg
     3661 gcggccttgc tgttcttcta cggcaaggtg ctgtgcacgg atctgccctg gcttcaggag
     3721 atcggaagac ctcggccgtc gcggcgcttg ccggtggtgc tgaccccgga tgaagtggtt
     3781 cgcatcctcg gttttctgga aggcgagcat cgtttgttcg cccaggactc tagctatagt
     3841 tctagtggtt ggctacgtat actccggaat attaatagat catggagata attaaaatga
     3901 taaccatctc gcaaataaat aagtatttta ctgttttcgt aacagttttg taataaaaaa
     3961 acctataaat attccggatt attcataccg tcccaccatc gggcgcggat cccggtccga
     4021 agcgcgcgga attcaaaggc ctacgtcgac gccatgcgcg cgcacccggg gggaggccgc
     4081 tgctgcccgg agcaggagga gggggagagc gcggcgggcg gcagcggcgc tggcggcgac
     4141 tccgccatag agcagggggg ccagggcagc gcgctcgctc cgtccccggt gagcggcgtg
     4201 cgcagggaag gcgctcgggg cggcggccgt ggccgggggc ggtggaagca ggcggcccgg
     4261 ggcggcggcg tctgtggccg tggccgtggc cgtggccggg gtcggggccg tggccggggc
     4321 cggggccggg gccgcggccg tccccagagt ggcggcagcg gccttggcgg cgacggcggc
     4381 ggcggcgcgg gcggctgcgg cgtcggcagc ggtggcggcg tcgccccccg gcgggatcct
     4441 gtccctttcc cgtcggggag ctcggggccg gggcccaggg gaccccgggc cacggagagc
     4501 gggaagagga tggactgccc ggccctcccc cccggatgga agaaggagga agtgatccga
     4561 aaatcagggc tcagtgctgg caagagcgat gtctactact tcagtccaag tggtaagaag
     4621 ttcagaagta aacctcagct ggcaagatac ctgggaaatg ctgttgacct tagcagtttt
     4681 gacttcagga ccggcaagat gatgcctagt aaattacaga agaacaagca gagactccgg
     4741 aatgaccccc tcaatcagaa caagggtaaa ccagacctga acacaacatt gccaattaga
     4801 caaactgcat caattttcaa gcaaccagta accaaattca cgaaccaccc gagcaataag
     4861 gtgaagtcag acccccagcg gatgaatgaa caaccacgtc agcttttctg ggagaagagg
     4921 ctacaaggac ttagcgcatc agatgtaaca gaacaaatta taaaaaccat ggagctacct
     4981 aaaggtcttc aaggagtcgg tccaggtagc aatgacgaga cccttctgtc tgctgtggcc
     5041 agtgctttac acacaagctc tgcgcccatc acaggacaag tctctgctgc cgtggaaaag
     5101 aaccctgctg tttggcttaa cacatctcaa cccctctgca aagctttcat tgttacagat
     5161 gaagacatta ggaaacagga agagcgagtc caacaagtac gcaagaaact ggaggaggca
     5221 ctgatggccg acatcctgtc ccgggctgcg gacacggagg aagtagacat tgacatggac
     5281 gatggagatg aggcgtgcgg ccgcgccatg gtgagcaagg gcgaggagct gttcaccggg
     5341 gtggtgccca tcctggtcga gctggacggc gacgtaaacg gccacaagtt cagcgtgtcc
     5401 ggcgagggcg agggcgatgc cacctacggc aagctgaccc tgaagttcat ctgcaccacc
     5461 ggcaagctgc ccgtgccctg gcccaccctc gtgaccaccc tgacctacgg cgtgcagtgc
     5521 ttcagccgct accccgacca catgaagcag cacgacttct tcaagtccgc catgcccgaa
     5581 ggctacgtcc aggagcgcac catcttcttc aaggacgacg gcaactacaa gacccgcgcc
     5641 gaggtgaagt tcgagggcga caccctggtg aaccgcatcg agctgaaggg catcgacttc
     5701 aaggaggacg gcaacatcct ggggcacaag ctggagtaca actacaacag ccacaacgtc
     5761 tatatcatgg ccgacaagca gaagaacggc atcaaggtga acttcaagat ccgccacaac
     5821 atcgaggacg gcagcgtgca gctcgccgac cactaccagc agaacacccc catcggcgac
     5881 ggccccgtgc tgctgcccga caaccactac ctgagcaccc agtccgccct gagcaaagac
     5941 cccaacgaga agcgcgatca catggtcctg ctggagttcg tgaccgccgc cgggatcact
     6001 ctcggcatgg acgagctgta caagtaatct agagcctgca gtctcgaggc atgcggtacc
     6061 aagcttgtcg agaagtacta gaggatcata atcagccata ccacatttgt agaggtttta
     6121 cttgctttaa aaaacctccc acacctcccc ctgaacctga aacataaaat gaatgcaatt
     6181 gttgttgtta acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca
     6241 aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc
     6301 aatgtatctt atcatgtctg gatctgatca ctgcttgagc ctaggagatc cgaaccagat
     6361 aagtgaaatc tagttccaaa ctattttgtc atttttaatt ttcgtattag cttacgacgc
     6421 tacacccagt tcccatctat tttgtcactc ttccctaaat aatccttaaa aactccattt
     6481 ccacccctcc cagttcccaa ctattttgtc cgcccacagc ggggcatttt tcttcctgtt
     6541 atgtttttaa tcaaacatcc tgccaactcc atgtgacaaa ccgtcatctt cggctacttt
     6601 ttctctgtca cagaatgaaa atttttctgt catctcttcg ttattaatgt ttgtaattga
     6661 ctgaatatca acgcttattt gcagcctgaa tggcgaatgg gacgcg
//
