LOCUS       sequence-422715-        5870 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses MeCP2 (aa163-309) GFP tagged in insect cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5870)
  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 5870)
  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-MeCP2G.9 (pc1574)
FEATURES             Location/Qualifiers
     source          1..5870
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      23..477
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(110..129)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     319..340
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        503..607
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(608..676,677..1468)
                     /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(826..845)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1639..2227
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2128..2147
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2381..2398
                     /label=L4440
                     /note="L4440 vector, forward primer"
     mobile_element  2532..2756
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7R
                     /note="mini-Tn7 element (right end of the Tn7 transposon)"
     CDS             complement(2823..3356)
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLWSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     promoter        complement(join(3526..3531,3532..3548,3549..3554))
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     promoter        3906..3997
                     /gene="polh from Autographa californica"
                     /label=polyhedrin promoter
                     /note="promoter for the baculovirus polyhedrin gene"
     primer_bind     3922..3939
                     /label=Polyhedrin-F
                     /note="Polyhedrin promoter, forward primer"
     CDS             join(4499..4501,4502..4504,4505..5218)
                     /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(4499..4501,4502..4504,4505..5215)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(4544..4565)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(4805..4824)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     5152..5173
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    5381..5515
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5418..5437)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5472..5491
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     mobile_element  5544..5709
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7L
                     /note="mini-Tn7 element (left end of the Tn7 transposon)"
ORIGIN
        1 tgcagcctga atggcgaatg ggacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg
       61 gtggttacgc gcagcgtgac cgctacactt gccagcgccc tagcgcccgc tcctttcgct
      121 ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggg
      181 ctccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgattag
      241 ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc tttgacgttg
      301 gagtccacgt tcttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct
      361 cggtctattc ttttgattta taagggattt tgccgatttc ggcctattgg ttaaaaaatg
      421 agctgattta acaaaaattt aacgcgaatt ttaacaaaat attaacgctt acaatttagg
      481 tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc
      541 aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag
      601 gaagagtatg agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg
      661 ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta aaagatgctg aagatcagtt
      721 gggtgcacga gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt
      781 tcgccccgaa gaacgttttc caatgatgag cacttttaaa gttctgctat gtggcgcggt
      841 attatcccgt attgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa
      901 tgacttggtt gagtactcac cagtcacaga aaagcatctt acggatggca tgacagtaag
      961 agaattatgc agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac
     1021 aacgatcgga ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac
     1081 tcgccttgat cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac
     1141 cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg aactacttac
     1201 tctagcttcc cggcaacaat taatagactg gatggaggcg gataaagttg caggaccact
     1261 tctgcgctcg gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg
     1321 tgggtctcgc ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt
     1381 tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat
     1441 aggtgcctca ctgattaagc attggtaact gtcagaccaa gtttactcat atatacttta
     1501 gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa
     1561 tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga
     1621 aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac
     1681 aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt
     1741 tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc
     1801 gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat
     1861 cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag
     1921 acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc
     1981 cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag
     2041 cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac
     2101 aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg
     2161 gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct
     2221 atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc
     2281 tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgcctttga
     2341 gtgagctgat accgctcgcc gcagccgaac gaccgagcgc agcgagtcag tgagcgagga
     2401 agcggaagag cgcctgatgc ggtattttct ccttacgcat ctgtgcggta tttcacaccg
     2461 catagaccag ccgcgtaacc tggcaaaatc ggttacggtt gagtaataaa tggatgccct
     2521 gcgtaagcgg gtgtgggcgg acaataaagt cttaaactga acaaaataga tctaaactat
     2581 gacaataaag tcttaaacta gacagaatag ttgtaaactg aaatcagtcc agttatgctg
     2641 tgaaaaagca tactggactt ttgttatggc taaagcaaac tcttcatttt ctgaagtgca
     2701 aattgcccgt cgtattaaag aggggcgtgg ccaagggcat ggtaaagact atattcgcgg
     2761 cgttgtgaca atttaccgaa caactccgcg gccgggaagc cgatctcggc ttgaacgaat
     2821 tgttaggtgg cggtacttgg gtcgatatca aagtgcatca cttcttcccg tatgcccaac
     2881 tttgtataga gagccactgc gggatcgtca ccgtaatctg cttgcacgta gatcacataa
     2941 gcaccaagcg cgttggcctc atgcttgagg agattgatga gcgcggtggc aatgccctgc
     3001 ctccggtgct cgccggagac tgcgagatca tagatataga tctcactacg cggctgctca
     3061 aacttgggca gaacgtaagc cgcgagagcg ccaacaaccg cttcttggtc gaaggcagca
     3121 agcgcgatga atgtcttact acggagcaag ttcccgaggt aatcggagtc cggctgatgt
     3181 tgggagtagg tggctacgtc tccgaactca cgaccgaaaa gatcaagagc agcccgcatg
     3241 gatttgactt ggtcagggcc gagcctacat gtgcgaatga tgcccatact tgagccacct
     3301 aactttgttt tagggcgact gccctgctgc gtaacatcgt tgctgctcca taacatcaaa
     3361 catcgaccca cggcgtaacg cgcttgctgc ttggatgccc gaggcataga ctgtacaaaa
     3421 aaacagtcat aacaagccat gaaaaccgcc actgcgccgt taccaccgct gcgttcggtc
     3481 aaggttctgg accagttgcg tgagcgcata cgctacttgc attacagttt acgaaccgaa
     3541 caggcttatg tcaactgggt tcgtgccttc atccgtttcc acggtgtgcg tcacccggca
     3601 accttgggca gcagcgaagt cgaggcattt ctgtcctggc tggcgaacga gcgcaaggtt
     3661 tcggtctcca cgcatcgtca ggcattggcg gccttgctgt tcttctacgg caaggtgctg
     3721 tgcacggatc tgccctggct tcaggagatc ggaagacctc ggccgtcgcg gcgcttgccg
     3781 gtggtgctga ccccggatga agtggttcgc atcctcggtt ttctggaagg cgagcatcgt
     3841 ttgttcgccc aggactctag ctatagttct agtggttggc tacgtatact ccggaatatt
     3901 aatagatcat ggagataatt aaaatgataa ccatctcgca aataaataag tattttactg
     3961 ttttcgtaac agttttgtaa taaaaaaacc tataaatatt ccggattatt cataccgtcc
     4021 caccatcggg cgcggatccg ccatggggag cccttccagg agagaacaga aaccacctaa
     4081 gaagcccaaa tctcccaaag ctccaggaac tggcaggggt cggggacgcc cgaaagggag
     4141 cggcactggg agaccaaagg cagcagcatc agaaggtgtt caagtgaaaa gggtcctgga
     4201 gaagagccct gggaaacttc tcgtcaagat gcctttccaa gcatcacctg ggggtaaggg
     4261 tgagggaggt ggggctacca catctgcgca ggtcatggtg atcaaacgcc ctggcagaaa
     4321 gcgaaaagct gaagctgacc cccaggccat tcctaagaaa cggggtagaa agcctgggag
     4381 tgtggtggca gctgctgctg cagaggccaa aaagaaagct gtgaaggaat cttctatacg
     4441 gtctgtgcag gagactgtgc tccccatcaa gaagcgcaag acccggagcg gccgcgccat
     4501 ggtgagcaag ggcgaggagc tgttcaccgg ggtggtgccc atcctggtcg agctggacgg
     4561 cgacgtaaac ggccacaagt tcagcgtgtc cggcgagggc gagggcgatg ccacctacgg
     4621 caagctgacc ctgaagttca tctgcaccac cggcaagctg cccgtgccct ggcccaccct
     4681 cgtgaccacc ctgacctacg gcgtgcagtg cttcagccgc taccccgacc acatgaagca
     4741 gcacgacttc ttcaagtccg ccatgcccga aggctacgtc caggagcgca ccatcttctt
     4801 caaggacgac ggcaactaca agacccgcgc cgaggtgaag ttcgagggcg acaccctggt
     4861 gaaccgcatc gagctgaagg gcatcgactt caaggaggac ggcaacatcc tggggcacaa
     4921 gctggagtac aactacaaca gccacaacgt ctatatcatg gccgacaagc agaagaacgg
     4981 catcaaggtg aacttcaaga tccgccacaa catcgaggac ggcagcgtgc agctcgccga
     5041 ccactaccag cagaacaccc ccatcggcga cggccccgtg ctgctgcccg acaaccacta
     5101 cctgagcacc cagtccgccc tgagcaaaga ccccaacgag aagcgcgatc acatggtcct
     5161 gctggagttc gtgaccgccg ccgggatcac tctcggcatg gacgagctgt acaagtaatc
     5221 tagagcctgc agtctcgagg catgcggtac caagcttgtc gagaagtact agaggatcat
     5281 aatcagccat accacatttg tagaggtttt acttgcttta aaaaacctcc cacacctccc
     5341 cctgaacctg aaacataaaa tgaatgcaat tgttgttgtt aacttgttta ttgcagctta
     5401 taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact
     5461 gcattctagt tgtggtttgt ccaaactcat caatgtatct tatcatgtct ggatctgatc
     5521 actgcttgag cctaggagat ccgaaccaga taagtgaaat ctagttccaa actattttgt
     5581 catttttaat tttcgtatta gcttacgacg ctacacccag ttcccatcta ttttgtcact
     5641 cttccctaaa taatccttaa aaactccatt tccacccctc ccagttccca actattttgt
     5701 ccgcccacag cggggcattt ttcttcctgt tatgttttta atcaaacatc ctgccaactc
     5761 catgtgacaa accgtcatct tcggctactt tttctctgtc acagaatgaa aatttttctg
     5821 tcatctcttc gttattaatg tttgtaattg actgaatatc aacgcttatt
//
