LOCUS       sequence-422713-        6050 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses MeCP2 (aa310-492) GFP tagged in insect cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6050)
  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 6050)
  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.8 (pc1573)
FEATURES             Location/Qualifiers
     source          1..6050
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(145..733)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(225..244)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(904..1695,1696..1764))
                     /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     1527..1546
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1765..1869)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(1895..2349)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2032..2053)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2243..2262
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     mobile_element  2533..2698
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7L
                     /note="mini-Tn7 element (left end of the Tn7 transposon)"
     polyA_signal    2727..2861
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2751..2770)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2805..2824
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(join(3039..3752,3753..3755,3756..3758))
                     /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             complement(join(3042..3752,3753..3755,3756..3758))
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(3084..3105)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     3433..3452
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3692..3713
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      3755..3764
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             complement(3952..3978)
                     /codon_start=1
                     /product="9xHis affinity tag"
                     /label=9xHis
                     /translation="HHHHHHHHH"
     CDS             complement(4188..4208)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=7xHis
                     /translation="HHHHHHH"
     promoter        complement(4425..4516)
                     /gene="polh from Autographa californica"
                     /label=polyhedrin promoter
                     /note="promoter for the baculovirus polyhedrin gene"
     primer_bind     complement(4483..4500)
                     /label=Polyhedrin-F
                     /note="Polyhedrin promoter, forward primer"
     promoter        join(4868..4873,4874..4890,4891..4896)
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     CDS             5066..5599
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLWSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     mobile_element  5666..5890
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7R
                     /note="mini-Tn7 element (right end of the Tn7 transposon)"
     primer_bind     complement(6024..6041)
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga
       61 atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg
      121 taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa
      181 aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt
      241 tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct
      301 gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct
      361 cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc
      421 cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt
      481 atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc
      541 tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat
      601 ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa
      661 acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa
      721 aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga
      781 aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct
      841 tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga
      901 cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc
      961 catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg
     1021 ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat
     1081 aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat
     1141 ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg
     1201 caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc
     1261 attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa
     1321 agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc
     1381 actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt
     1441 ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag
     1501 ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt
     1561 gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag
     1621 atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac
     1681 cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc
     1741 gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca
     1801 gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg
     1861 ggttccgcgc acatttcccc gaaaagtgcc acctaaattg taagcgttaa tattttgtta
     1921 aaattcgcgt taaatttttg ttaaatcagc tcatttttta accaataggc cgaaatcggc
     1981 aaaatccctt ataaatcaaa agaatagacc gagatagggt tgagtgttgt tccagtttgg
     2041 aacaagagtc cactattaag aacgtggact ccaacgtcaa agggcgaaaa accgtctatc
     2101 agggcgatgg cccactacgt gaaccatcac cctaatcaag ttttttgggg tcgaggtgcc
     2161 gtaaagcact aaatcggaac cctaaaggga gcccccgatt tagagcttga cggggaaagc
     2221 cggcgaacgt ggcgagaaag gaagggaaga aagcgaaagg agcgggcgct agggcgctgg
     2281 caagtgtagc ggtcacgctg cgcgtaacca ccacacccgc cgcgcttaat gcgccgctac
     2341 agggcgcgtc ccattcgcca ttcaggctgc aaataagcgt tgatattcag tcaattacaa
     2401 acattaataa cgaagagatg acagaaaaat tttcattctg tgacagagaa aaagtagccg
     2461 aagatgacgg tttgtcacat ggagttggca ggatgtttga ttaaaaacat aacaggaaga
     2521 aaaatgcccc gctgtgggcg gacaaaatag ttgggaactg ggaggggtgg aaatggagtt
     2581 tttaaggatt atttagggaa gagtgacaaa atagatggga actgggtgta gcgtcgtaag
     2641 ctaatacgaa aattaaaaat gacaaaatag tttggaacta gatttcactt atctggttcg
     2701 gatctcctag gctcaagcag tgatcagatc cagacatgat aagatacatt gatgagtttg
     2761 gacaaaccac aactagaatg cagtgaaaaa aatgctttat ttgtgaaatt tgtgatgcta
     2821 ttgctttatt tgtaaccatt ataagctgca ataaacaagt taacaacaac aattgcattc
     2881 attttatgtt tcaggttcag ggggaggtgt gggaggtttt ttaaagcaag taaaacctct
     2941 acaaatgtgg tatggctgat tatgatcctc tagtacttct cgacaagctt ggtaccgcat
     3001 gcctcgagac tgcaggctct agattcgaaa gcggccgctt acttgtacag ctcgtccatg
     3061 ccgagagtga tcccggcggc ggtcacgaac tccagcagga ccatgtgatc gcgcttctcg
     3121 ttggggtctt tgctcagggc ggactgggtg ctcaggtagt ggttgtcggg cagcagcacg
     3181 gggccgtcgc cgatgggggt gttctgctgg tagtggtcgg cgagctgcac gctgccgtcc
     3241 tcgatgttgt ggcggatctt gaagttcacc ttgatgccgt tcttctgctt gtcggccatg
     3301 atatagacgt tgtggctgtt gtagttgtac tccagcttgt gccccaggat gttgccgtcc
     3361 tccttgaagt cgatgccctt cagctcgatg cggttcacca gggtgtcgcc ctcgaacttc
     3421 acctcggcgc gggtcttgta gttgccgtcg tccttgaaga agatggtgcg ctcctggacg
     3481 tagccttcgg gcatggcgga cttgaagaag tcgtgctgct tcatgtggtc ggggtagcgg
     3541 ctgaagcact gcacgccgta ggtcagggtg gtcacgaggg tgggccaggg cacgggcagc
     3601 ttgccggtgg tgcagatgaa cttcagggtc agcttgccgt aggtggcatc gccctcgccc
     3661 tcgccggaca cgctgaactt gtggccgttt acgtcgccgt ccagctcgac caggatgggc
     3721 accaccccgg tgaacagctc ctcgcccttg ctcaccatgg tggcgaccgg tggatcccgg
     3781 gcccgcggta ccgtcgacgg tatcgataag cttgatatcg aattcctgct aactctctcg
     3841 gtcacgggcg tccggctgtc cacaggctcc tctctgtttg gcctcggcat ggaggatgaa
     3901 acaatgtctt tgcgctctcc ctcccctcgg tgtttgtact tttctgcaac tgtggtggtg
     3961 gtggtggtgg tggtggtggc ggcggcagca accatgggct gagtcttagc tggctccttg
     4021 gggcagccat cgctttccag tgagcctgct cggggcatct tctcctcttt gcagatgctg
     4081 ctgctcaagt cctgaggctc aggggggctg atggggtcct cagagctctg aggctcaggt
     4141 gggggtggag gtggaagcaa tggcatgggg gcctttgggg actctgcgtg atggtggtga
     4201 tgatggtgct ccttcttagg tggtgaggag gcactgctgc tgcgcccctt ggggctgctc
     4261 tccttgcttt tacgcccagg gctcttgcat gtcttcagtc cctttccact cttctcacca
     4321 agtgtagaca ccagcagggg cttcaccacc tccttgacct caatgctgac ggtttccatg
     4381 gcagatccgc gcccgatggt gggacggtat gaataatccg gaatatttat aggttttttt
     4441 attacaaaac tgttacgaaa acagtaaaat acttatttat ttgcgagatg gttatcattt
     4501 taattatctc catgatctat taatattccg gagtatacgt agccaaccac tagaactata
     4561 gctagagtcc tgggcgaaca aacgatgctc gccttccaga aaaccgagga tgcgaaccac
     4621 ttcatccggg gtcagcacca ccggcaagcg ccgcgacggc cgaggtcttc cgatctcctg
     4681 aagccagggc agatccgtgc acagcacctt gccgtagaag aacagcaagg ccgccaatgc
     4741 ctgacgatgc gtggagaccg aaaccttgcg ctcgttcgcc agccaggaca gaaatgcctc
     4801 gacttcgctg ctgcccaagg ttgccgggtg acgcacaccg tggaaacgga tgaaggcacg
     4861 aacccagttg acataagcct gttcggttcg taaactgtaa tgcaagtagc gtatgcgctc
     4921 acgcaactgg tccagaacct tgaccgaacg cagcggtggt aacggcgcag tggcggtttt
     4981 catggcttgt tatgactgtt tttttgtaca gtctatgcct cgggcatcca agcagcaagc
     5041 gcgttacgcc gtgggtcgat gtttgatgtt atggagcagc aacgatgtta cgcagcaggg
     5101 cagtcgccct aaaacaaagt taggtggctc aagtatgggc atcattcgca catgtaggct
     5161 cggccctgac caagtcaaat ccatgcgggc tgctcttgat cttttcggtc gtgagttcgg
     5221 agacgtagcc acctactccc aacatcagcc ggactccgat tacctcggga acttgctccg
     5281 tagtaagaca ttcatcgcgc ttgctgcctt cgaccaagaa gcggttgttg gcgctctcgc
     5341 ggcttacgtt ctgcccaagt ttgagcagcc gcgtagtgag atctatatct atgatctcgc
     5401 agtctccggc gagcaccgga ggcagggcat tgccaccgcg ctcatcaatc tcctcaagca
     5461 tgaggccaac gcgcttggtg cttatgtgat ctacgtgcaa gcagattacg gtgacgatcc
     5521 cgcagtggct ctctatacaa agttgggcat acgggaagaa gtgatgcact ttgatatcga
     5581 cccaagtacc gccacctaac aattcgttca agccgagatc ggcttcccgg ccgcggagtt
     5641 gttcggtaaa ttgtcacaac gccgcgaata tagtctttac catgcccttg gccacgcccc
     5701 tctttaatac gacgggcaat ttgcacttca gaaaatgaag agtttgcttt agccataaca
     5761 aaagtccagt atgctttttc acagcataac tggactgatt tcagtttaca actattctgt
     5821 ctagtttaag actttattgt catagtttag atctattttg ttcagtttaa gactttattg
     5881 tccgcccaca cccgcttacg cagggcatcc atttattact caaccgtaac cgattttgcc
     5941 aggttacgcg gctggtctat gcggtgtgaa ataccgcaca gatgcgtaag gagaaaatac
     6001 cgcatcaggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt
//
