LOCUS       sequence-422697-        6014 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses MeCP2 (aa1-163) YFP tagged in insect cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6014)
  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 6014)
  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-MeCP2Y.3 (pc1575)
FEATURES             Location/Qualifiers
     source          1..6014
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      10..464
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(97..116)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     306..327
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        490..594
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(595..663,664..1455)
                     /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(813..832)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1626..2214
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2115..2134
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2368..2385
                     /label=L4440
                     /note="L4440 vector, forward primer"
     mobile_element  2519..2743
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7R
                     /note="mini-Tn7 element (right end of the Tn7 transposon)"
     CDS             complement(2810..3343)
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLWSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     promoter        complement(join(3513..3518,3519..3535,3536..3541))
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     promoter        3893..3984
                     /gene="polh from Autographa californica"
                     /label=polyhedrin promoter
                     /note="promoter for the baculovirus polyhedrin gene"
     primer_bind     3909..3926
                     /label=Polyhedrin-F
                     /note="Polyhedrin promoter, forward primer"
     regulatory      4608..4617
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(4614..4616,4617..4619,4620..5333)
                     /codon_start=1
                     /product="enhanced YFP"
                     /label=EYFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(4659..4680)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(4920..4939)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     5267..5288
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    5512..5646
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5549..5568)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5603..5622
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     mobile_element  5675..5840
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7L
                     /note="mini-Tn7 element (left end of the Tn7 transposon)"
ORIGIN
        1 gcgaatggga cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca
       61 gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc tttcgctttc ttcccttcct
      121 ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcgggggctc cctttagggt
      181 tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgattagggt gatggttcac
      241 gtagtgggcc atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct
      301 taatagtgga ctcttgttcc aaactggaac aacactcaac cctatctcgg tctattcttt
      361 tgatttataa gggattttgc cgatttcggc ctattggtta aaaaatgagc tgatttaaca
      421 aaaatttaac gcgaatttta acaaaatatt aacgcttaca atttaggtgg cacttttcgg
      481 ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg
      541 ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtatgagt
      601 attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt
      661 gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg
      721 ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa
      781 cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt
      841 gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag
      901 tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt
      961 gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac gatcggagga
     1021 ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt
     1081 tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgta
     1141 gcaatggcaa caacgttgcg caaactatta actggcgaac tacttactct agcttcccgg
     1201 caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc
     1261 cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt
     1321 atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg
     1381 gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg
     1441 attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat tgatttaaaa
     1501 cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa
     1561 atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga
     1621 tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg
     1681 ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact
     1741 ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac
     1801 cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg
     1861 gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg
     1921 gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga
     1981 acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc
     2041 gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg
     2101 agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc
     2161 tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc
     2221 agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt
     2281 cctgcgttat cccctgattc tgtggataac cgtattaccg cctttgagtg agctgatacc
     2341 gctcgccgca gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc ggaagagcgc
     2401 ctgatgcggt attttctcct tacgcatctg tgcggtattt cacaccgcat agaccagccg
     2461 cgtaacctgg caaaatcggt tacggttgag taataaatgg atgccctgcg taagcgggtg
     2521 tgggcggaca ataaagtctt aaactgaaca aaatagatct aaactatgac aataaagtct
     2581 taaactagac agaatagttg taaactgaaa tcagtccagt tatgctgtga aaaagcatac
     2641 tggacttttg ttatggctaa agcaaactct tcattttctg aagtgcaaat tgcccgtcgt
     2701 attaaagagg ggcgtggcca agggcatggt aaagactata ttcgcggcgt tgtgacaatt
     2761 taccgaacaa ctccgcggcc gggaagccga tctcggcttg aacgaattgt taggtggcgg
     2821 tacttgggtc gatatcaaag tgcatcactt cttcccgtat gcccaacttt gtatagagag
     2881 ccactgcggg atcgtcaccg taatctgctt gcacgtagat cacataagca ccaagcgcgt
     2941 tggcctcatg cttgaggaga ttgatgagcg cggtggcaat gccctgcctc cggtgctcgc
     3001 cggagactgc gagatcatag atatagatct cactacgcgg ctgctcaaac ttgggcagaa
     3061 cgtaagccgc gagagcgcca acaaccgctt cttggtcgaa ggcagcaagc gcgatgaatg
     3121 tcttactacg gagcaagttc ccgaggtaat cggagtccgg ctgatgttgg gagtaggtgg
     3181 ctacgtctcc gaactcacga ccgaaaagat caagagcagc ccgcatggat ttgacttggt
     3241 cagggccgag cctacatgtg cgaatgatgc ccatacttga gccacctaac tttgttttag
     3301 ggcgactgcc ctgctgcgta acatcgttgc tgctccataa catcaaacat cgacccacgg
     3361 cgtaacgcgc ttgctgcttg gatgcccgag gcatagactg tacaaaaaaa cagtcataac
     3421 aagccatgaa aaccgccact gcgccgttac caccgctgcg ttcggtcaag gttctggacc
     3481 agttgcgtga gcgcatacgc tacttgcatt acagtttacg aaccgaacag gcttatgtca
     3541 actgggttcg tgccttcatc cgtttccacg gtgtgcgtca cccggcaacc ttgggcagca
     3601 gcgaagtcga ggcatttctg tcctggctgg cgaacgagcg caaggtttcg gtctccacgc
     3661 atcgtcaggc attggcggcc ttgctgttct tctacggcaa ggtgctgtgc acggatctgc
     3721 cctggcttca ggagatcgga agacctcggc cgtcgcggcg cttgccggtg gtgctgaccc
     3781 cggatgaagt ggttcgcatc ctcggttttc tggaaggcga gcatcgtttg ttcgcccagg
     3841 actctagcta tagttctagt ggttggctac gtatactccg gaatattaat agatcatgga
     3901 gataattaaa atgataacca tctcgcaaat aaataagtat tttactgttt tcgtaacagt
     3961 tttgtaataa aaaaacctat aaatattccg gattattcat accgtcccac catcgggcgc
     4021 ggatcccggt ccgaagcgcg cggaattcaa aggcctacgt cgacgagctc actagtcgcg
     4081 gccgctctag catggtagct gggatgttag ggctcaggaa ggaaaagtca gaagaccagg
     4141 atctccaggg cctcaaagag aaacccctga agtttaagaa ggtgaagaaa gacaagaagg
     4201 aagacaaaga gggcaaacat gaaccactac agccttcagc ccaccattct gcagagccag
     4261 cagaggcagg caaagcagag acatcagaaa gctcaggctc tgccccagca gtaccagaag
     4321 cctctgcttc tcccaaacag cgacgttcca tcattcgtga ccggggacct atgtatgatg
     4381 accccacctt gcctgaaggt tggacgcgaa agcttaaaca gaggaagtct ggtcgctctg
     4441 ctggaaagta tgatgtatat ttgatcaatc cccagggaaa agcctttcgc tctaaagtag
     4501 aattgattgc atattttgaa aaggtgggag acacctcctt ggaccctaat gattttgact
     4561 tcactgtaac tgggagaggg gtaccgcggg cccgggatcc accggtcgcc accatggtga
     4621 gcaagggcga ggagctgttc accggggtgg tgcccatcct ggtcgagctg gacggcgacg
     4681 taaacggcca caagttcagc gtgtccggcg agggcgaggg cgatgccacc tacggcaagc
     4741 tgaccctgaa gttcatctgc accaccggca agctgcccgt gccctggccc accctcgtga
     4801 ccaccttcgg ctacggcctg cagtgcttcg cccgctaccc cgaccacatg aagcagcacg
     4861 acttcttcaa gtccgccatg cccgaaggct acgtccagga gcgcaccatc ttcttcaagg
     4921 acgacggcaa ctacaagacc cgcgccgagg tgaagttcga gggcgacacc ctggtgaacc
     4981 gcatcgagct gaagggcatc gacttcaagg aggacggcaa catcctgggg cacaagctgg
     5041 agtacaacta caacagccac aacgtctata tcatggccga caagcagaag aacggcatca
     5101 aggtgaactt caagatccgc cacaacatcg aggacggcag cgtgcagctc gccgaccact
     5161 accagcagaa cacccccatc ggcgacggcc ccgtgctgct gcccgacaac cactacctga
     5221 gctaccagtc cgccctgagc aaagacccca acgagaagcg cgatcacatg gtcctgctgg
     5281 agttcgtgac cgccgccggg atcactctcg gcatggacga gctgtacaag taaagcggcc
     5341 gctttcgaat ctagagcctg cagtctcgag gcatgcggta ccaagcttgt cgagaagtac
     5401 tagaggatca taatcagcca taccacattt gtagaggttt tacttgcttt aaaaaacctc
     5461 ccacacctcc ccctgaacct gaaacataaa atgaatgcaa ttgttgttgt taacttgttt
     5521 attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca
     5581 tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc
     5641 tggatctgat cactgcttga gcctaggaga tccgaaccag ataagtgaaa tctagttcca
     5701 aactattttg tcatttttaa ttttcgtatt agcttacgac gctacaccca gttcccatct
     5761 attttgtcac tcttccctaa ataatcctta aaaactccat ttccacccct cccagttccc
     5821 aactattttg tccgcccaca gcggggcatt tttcttcctg ttatgttttt aatcaaacat
     5881 cctgccaact ccatgtgaca aaccgtcatc ttcggctact ttttctctgt cacagaatga
     5941 aaatttttct gtcatctctt cgttattaat gtttgtaatt gactgaatat caacgcttat
     6001 ttgcagcctg aatg
//
