LOCUS       sequence-422688-        7126 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses fl. MBD4 GFP tagged in insect cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7126)
  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 7126)
  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: pFBMBD4-C-GFP (pc1567)
FEATURES             Location/Qualifiers
     source          1..7126
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      24..478
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(111..130)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     320..341
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        504..608
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(609..677,678..1469)
                     /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(827..846)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1640..2228
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2129..2148
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2382..2399
                     /label=L4440
                     /note="L4440 vector, forward primer"
     mobile_element  2533..2757
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7R
                     /note="mini-Tn7 element (right end of the Tn7 transposon)"
     CDS             complement(2824..3357)
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLWSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     promoter        complement(join(3527..3532,3533..3549,3550..3555))
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     promoter        3907..3998
                     /gene="polh from Autographa californica"
                     /label=polyhedrin promoter
                     /note="promoter for the baculovirus polyhedrin gene"
     primer_bind     3923..3940
                     /label=Polyhedrin-F
                     /note="Polyhedrin promoter, forward primer"
     CDS             5089..5100
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     CDS             join(5756..5758,5759..5761,5762..6475)
                     /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(5756..5758,5759..5761,5762..6472)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(5801..5822)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(6062..6081)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     6409..6430
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    6638..6772
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6675..6694)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6729..6748
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     mobile_element  6801..6966
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7L
                     /note="mini-Tn7 element (left end of the Tn7 transposon)"
ORIGIN
        1 ttgcagcctg aatggcgaat gggacgcgcc ctgtagcggc gcattaagcg cggcgggtgt
       61 ggtggttacg cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc
      121 tttcttccct tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg
      181 gctcccttta gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgatta
      241 gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt
      301 ggagtccacg ttcttaatag tggactcttg ttccaaactg gaacaacact caaccctatc
      361 tcggtctatt cttttgattt ataagggatt ttgccgattt cggcctattg gttaaaaaat
      421 gagctgattt aacaaaaatt taacgcgaat tttaacaaaa tattaacgct tacaatttag
      481 gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt
      541 caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa
      601 ggaagagtat gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt
      661 gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt
      721 tgggtgcacg agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt
      781 ttcgccccga agaacgtttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg
      841 tattatcccg tattgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga
      901 atgacttggt tgagtactca ccagtcacag aaaagcatct tacggatggc atgacagtaa
      961 gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga
     1021 caacgatcgg aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa
     1081 ctcgccttga tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca
     1141 ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact attaactggc gaactactta
     1201 ctctagcttc ccggcaacaa ttaatagact ggatggaggc ggataaagtt gcaggaccac
     1261 ttctgcgctc ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc
     1321 gtgggtctcg cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag
     1381 ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga
     1441 taggtgcctc actgattaag cattggtaac tgtcagacca agtttactca tatatacttt
     1501 agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata
     1561 atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag
     1621 aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa
     1681 caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt
     1741 ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc
     1801 cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa
     1861 tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa
     1921 gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc
     1981 ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa
     2041 gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa
     2101 caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg
     2161 ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc
     2221 tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg
     2281 ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg
     2341 agtgagctga taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg
     2401 aagcggaaga gcgcctgatg cggtattttc tccttacgca tctgtgcggt atttcacacc
     2461 gcatagacca gccgcgtaac ctggcaaaat cggttacggt tgagtaataa atggatgccc
     2521 tgcgtaagcg ggtgtgggcg gacaataaag tcttaaactg aacaaaatag atctaaacta
     2581 tgacaataaa gtcttaaact agacagaata gttgtaaact gaaatcagtc cagttatgct
     2641 gtgaaaaagc atactggact tttgttatgg ctaaagcaaa ctcttcattt tctgaagtgc
     2701 aaattgcccg tcgtattaaa gaggggcgtg gccaagggca tggtaaagac tatattcgcg
     2761 gcgttgtgac aatttaccga acaactccgc ggccgggaag ccgatctcgg cttgaacgaa
     2821 ttgttaggtg gcggtacttg ggtcgatatc aaagtgcatc acttcttccc gtatgcccaa
     2881 ctttgtatag agagccactg cgggatcgtc accgtaatct gcttgcacgt agatcacata
     2941 agcaccaagc gcgttggcct catgcttgag gagattgatg agcgcggtgg caatgccctg
     3001 cctccggtgc tcgccggaga ctgcgagatc atagatatag atctcactac gcggctgctc
     3061 aaacttgggc agaacgtaag ccgcgagagc gccaacaacc gcttcttggt cgaaggcagc
     3121 aagcgcgatg aatgtcttac tacggagcaa gttcccgagg taatcggagt ccggctgatg
     3181 ttgggagtag gtggctacgt ctccgaactc acgaccgaaa agatcaagag cagcccgcat
     3241 ggatttgact tggtcagggc cgagcctaca tgtgcgaatg atgcccatac ttgagccacc
     3301 taactttgtt ttagggcgac tgccctgctg cgtaacatcg ttgctgctcc ataacatcaa
     3361 acatcgaccc acggcgtaac gcgcttgctg cttggatgcc cgaggcatag actgtacaaa
     3421 aaaacagtca taacaagcca tgaaaaccgc cactgcgccg ttaccaccgc tgcgttcggt
     3481 caaggttctg gaccagttgc gtgagcgcat acgctacttg cattacagtt tacgaaccga
     3541 acaggcttat gtcaactggg ttcgtgcctt catccgtttc cacggtgtgc gtcacccggc
     3601 aaccttgggc agcagcgaag tcgaggcatt tctgtcctgg ctggcgaacg agcgcaaggt
     3661 ttcggtctcc acgcatcgtc aggcattggc ggccttgctg ttcttctacg gcaaggtgct
     3721 gtgcacggat ctgccctggc ttcaggagat cggaagacct cggccgtcgc ggcgcttgcc
     3781 ggtggtgctg accccggatg aagtggttcg catcctcggt tttctggaag gcgagcatcg
     3841 tttgttcgcc caggactcta gctatagttc tagtggttgg ctacgtatac tccggaatat
     3901 taatagatca tggagataat taaaatgata accatctcgc aaataaataa gtattttact
     3961 gttttcgtaa cagttttgta ataaaaaaac ctataaatat tccggattat tcataccgtc
     4021 ccaccatcgg gcgcggatcc cggtccgaag cgcgcggaat tcaaaggcct acgtcgacgc
     4081 catggagagc ccaaaccttg gggacaacag agtccgtgga gagagcctag ttccagaccc
     4141 gccttgggat cgctgcaagg aagatattgc tgttggactg ggaggagtgg gagaagatgg
     4201 aaaggacctg gtgataagca gtgagcgcag ctcccttctc caagagccca ctgcttctac
     4261 tctgtctagt actacagcga cagaaggcca caagcctgtc ccgtgtggat gggaaagagt
     4321 tgtgaagcaa aggttatctg ggaaaactgc aggaaaattt gatgtatact ttatcagccc
     4381 acaaggattg aagttcagat caaaacgttc acttgctaat tatcttctca aaaatgggga
     4441 gacttttctt aagcctgaag attttaattt tactgtactg ccgaaaggga gcatcaatcc
     4501 cggttataaa caccaaagtt tggcagctct gacttccctg cagccaaatg aaactgacgt
     4561 ttcaaagcag aacctcaaga cacgaagcaa gtggaaaaca gatgtgttgc ctctgcccag
     4621 tggtacttca gagtcgccag aaagcagcgg actgtctaac tctaactcgg cttgcttgct
     4681 attgagagaa catagggaca ttcaggatgt tgactctgag aagaggagaa agtccaaaag
     4741 aaaggtgact gttttgaaag gaactgcaag tcagaaaacc aaacaaaagt gcaggaagag
     4801 tctcttagag tctactcaaa gaaacagaaa aagagcatct gtggttcaga aggtgggtgc
     4861 tgatcgcgag ctggtgccac aggaaagtca actcaacaga accctctgcc ctgcagatgc
     4921 ctgtgcaagg gagactgttg gcctggctgg ggaagaaaaa tcaccaagcc caggactgga
     4981 tctttgtttc atacaagtaa cttctggcac cacaaacaaa ttccattcaa ctgaagcagc
     5041 aggtgaagca aatcgtgagc agactttttt agaatcagag gaaatcagat cgaagggaga
     5101 cagaaagggg gaggcacatt tgcatactgg tgttttacag gatggctctg aaatgcccag
     5161 ctgctcacaa gccaagaaac actttacttc tgagacattt caagaagaca gcatcccacg
     5221 gacacaagta gaaaaaagga aaacaagcct gtatttttcc agcaagtaca acaaagaagc
     5281 tcttagcccc ccaagacgca aatccttcaa gaaatggacc cctcctcggt caccttttaa
     5341 tcttgttcaa gaaatacttt tccatgaccc atggaagctc ctcatcgcga ctatatttct
     5401 caatcggacc tcaggcaaga tggccatccc tgtgctgtgg gagtttctag agaagtaccc
     5461 ttcagctgaa gtggcccgag ctgccgactg gagggacgtg tcggagcttc tcaagcctct
     5521 tggtctctac gatctccgtg caaaaaccat tatcaagttc tcagatgaat atctgacaaa
     5581 gcagtggagg tatccgattg agcttcatgg cattggtaaa tatggcaacg actcctaccg
     5641 gatcttttgt gtcaatgaat ggaagcaggt gcaccctgaa gaccacaagt taaataaata
     5701 ccatgactgg ctttgggaga atcatgaaaa attaagtcta tcttgcggcc gcgccatggt
     5761 gagcaagggc gaggagctgt tcaccggggt ggtgcccatc ctggtcgagc tggacggcga
     5821 cgtaaacggc cacaagttca gcgtgtccgg cgagggcgag ggcgatgcca cctacggcaa
     5881 gctgaccctg aagttcatct gcaccaccgg caagctgccc gtgccctggc ccaccctcgt
     5941 gaccaccctg acctacggcg tgcagtgctt cagccgctac cccgaccaca tgaagcagca
     6001 cgacttcttc aagtccgcca tgcccgaagg ctacgtccag gagcgcacca tcttcttcaa
     6061 ggacgacggc aactacaaga cccgcgccga ggtgaagttc gagggcgaca ccctggtgaa
     6121 ccgcatcgag ctgaagggca tcgacttcaa ggaggacggc aacatcctgg ggcacaagct
     6181 ggagtacaac tacaacagcc acaacgtcta tatcatggcc gacaagcaga agaacggcat
     6241 caaggtgaac ttcaagatcc gccacaacat cgaggacggc agcgtgcagc tcgccgacca
     6301 ctaccagcag aacaccccca tcggcgacgg ccccgtgctg ctgcccgaca accactacct
     6361 gagcacccag tccgccctga gcaaagaccc caacgagaag cgcgatcaca tggtcctgct
     6421 ggagttcgtg accgccgccg ggatcactct cggcatggac gagctgtaca agtaatctag
     6481 agcctgcagt ctcgaggcat gcggtaccaa gcttgtcgag aagtactaga ggatcataat
     6541 cagccatacc acatttgtag aggttttact tgctttaaaa aacctcccac accttcccct
     6601 gaacctgaaa cataaaatga atgcaattgt tgttgttaac ttgtttattg cagcttataa
     6661 tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca
     6721 ttctagttgt ggtttgtcca aactcatcaa tgtatcttat catgtctgga tctgatcact
     6781 gcttgagcct aggagatccg aaccagataa gtgaaatcta gttccaaact attttgtcat
     6841 ttttaatttt cgtattagct tacgacgcta cacccagttc ccatctattt tgtcactctt
     6901 ccctaaataa tccttaaaaa ctccatttcc acccctccca gttcccaact attttgtccg
     6961 cccacagcgg ggcatttttc ttcctgttat gtttttaatc aaacatcctg ccaactccat
     7021 gtgacaaacc gtcatcttcg gctacttttt ctctgtcaca gaatgaaaat ttttctgtca
     7081 tctcttcgtt attaatgttt gtaattgact gaatatcaac gcttat
//
