LOCUS       sequence-422718-        5903 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses MeCP2 (aa163-309) mCherry tagged in insect cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5903)
  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 5903)
  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-MeCP2.9Ch (pc2385)
FEATURES             Location/Qualifiers
     source          1..5903
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      13..467
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(100..119)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     309..330
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        493..597
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(598..666,667..1458)
                     /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(816..835)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1629..2217
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2118..2137
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2371..2388
                     /label=L4440
                     /note="L4440 vector, forward primer"
     mobile_element  2522..2746
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7R
                     /note="mini-Tn7 element (right end of the Tn7 transposon)"
     CDS             complement(2813..3346)
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLWSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     promoter        complement(join(3516..3521,3522..3538,3539..3544))
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     promoter        3896..3987
                     /gene="polh from Autographa californica"
                     /label=polyhedrin promoter
                     /note="promoter for the baculovirus polyhedrin gene"
     primer_bind     3912..3929
                     /label=Polyhedrin-F
                     /note="Polyhedrin promoter, forward primer"
     CDS             4492..4512
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             4519..5229
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(4664..4682)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     4932..4951
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     polyA_signal    5404..5538
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5441..5460)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5495..5514
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     mobile_element  5567..5732
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7L
                     /note="mini-Tn7 element (left end of the Tn7 transposon)"
ORIGIN
        1 atggcgaatg ggacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc
       61 gcagcgtgac cgctacactt gccagcgccc tagcgcccgc tcctttcgct ttcttccctt
      121 cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggg ctccctttag
      181 ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgattag ggtgatggtt
      241 cacgtagtgg gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt
      301 tcttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc
      361 ttttgattta taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta
      421 acaaaaattt aacgcgaatt ttaacaaaat attaacgctt acaatttagg tggcactttt
      481 cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat
      541 ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtatg
      601 agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt
      661 tttgctcacc cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcacga
      721 gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa
      781 gaacgttttc caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt
      841 attgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt
      901 gagtactcac cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc
      961 agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac aacgatcgga
     1021 ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat
     1081 cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct
     1141 gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc
     1201 cggcaacaat taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg
     1261 gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc
     1321 ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg
     1381 acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca
     1441 ctgattaagc attggtaact gtcagaccaa gtttactcat atatacttta gattgattta
     1501 aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc
     1561 aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa
     1621 ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca
     1681 ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta
     1741 actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc
     1801 caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca
     1861 gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta
     1921 ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag
     1981 cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt
     2041 cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc
     2101 acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac
     2161 ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac
     2221 gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc
     2281 tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgcctttga gtgagctgat
     2341 accgctcgcc gcagccgaac gaccgagcgc agcgagtcag tgagcgagga agcggaagag
     2401 cgcctgatgc ggtattttct ccttacgcat ctgtgcggta tttcacaccg catagaccag
     2461 ccgcgtaacc tggcaaaatc ggttacggtt gagtaataaa tggatgccct gcgtaagcgg
     2521 gtgtgggcgg acaataaagt cttaaactga acaaaataga tctaaactat gacaataaag
     2581 tcttaaacta gacagaatag ttgtaaactg aaatcagtcc agttatgctg tgaaaaagca
     2641 tactggactt ttgttatggc taaagcaaac tcttcatttt ctgaagtgca aattgcccgt
     2701 cgtattaaag aggggcgtgg ccaagggcat ggtaaagact atattcgcgg cgttgtgaca
     2761 atttaccgaa caactccgcg gccgggaagc cgatctcggc ttgaacgaat tgttaggtgg
     2821 cggtacttgg gtcgatatca aagtgcatca cttcttcccg tatgcccaac tttgtataga
     2881 gagccactgc gggatcgtca ccgtaatctg cttgcacgta gatcacataa gcaccaagcg
     2941 cgttggcctc atgcttgagg agattgatga gcgcggtggc aatgccctgc ctccggtgct
     3001 cgccggagac tgcgagatca tagatataga tctcactacg cggctgctca aacttgggca
     3061 gaacgtaagc cgcgagagcg ccaacaaccg cttcttggtc gaaggcagca agcgcgatga
     3121 atgtcttact acggagcaag ttcccgaggt aatcggagtc cggctgatgt tgggagtagg
     3181 tggctacgtc tccgaactca cgaccgaaaa gatcaagagc agcccgcatg gatttgactt
     3241 ggtcagggcc gagcctacat gtgcgaatga tgcccatact tgagccacct aactttgttt
     3301 tagggcgact gccctgctgc gtaacatcgt tgctgctcca taacatcaaa catcgaccca
     3361 cggcgtaacg cgcttgctgc ttggatgccc gaggcataga ctgtacaaaa aaacagtcat
     3421 aacaagccat gaaaaccgcc actgcgccgt taccaccgct gcgttcggtc aaggttctgg
     3481 accagttgcg tgagcgcata cgctacttgc attacagttt acgaaccgaa caggcttatg
     3541 tcaactgggt tcgtgccttc atccgtttcc acggtgtgcg tcacccggca accttgggca
     3601 gcagcgaagt cgaggcattt ctgtcctggc tggcgaacga gcgcaaggtt tcggtctcca
     3661 cgcatcgtca ggcattggcg gccttgctgt tcttctacgg caaggtgctg tgcacggatc
     3721 tgccctggct tcaggagatc ggaagacctc ggccgtcgcg gcgcttgccg gtggtgctga
     3781 ccccggatga agtggttcgc atcctcggtt ttctggaagg cgagcatcgt ttgttcgccc
     3841 aggactctag ctatagttct agtggttggc tacgtatact ccggaatatt aatagatcat
     3901 ggagataatt aaaatgataa ccatctcgca aataaataag tattttactg ttttcgtaac
     3961 agttttgtaa taaaaaaacc tataaatatt ccggattatt cataccgtcc caccatcggg
     4021 cgcggatccg ccatggggag cccttccagg agagaacaga aaccacctaa gaagcccaaa
     4081 tctcccaaag ctccaggaac tggcaggggt cggggacgcc cgaaagggag cggcactggg
     4141 agaccaaagg cagcagcatc agaaggtgtt caagtgaaaa gggtcctgga gaagagccct
     4201 gggaaacttc tcgtcaagat gcctttccaa gcatcacctg ggggtaaggg tgagggaggt
     4261 ggggctacca catctgcgca ggtcatggtg atcaaacgcc ctggcagaaa gcgaaaagct
     4321 gaagctgacc cccaggccat tcctaagaaa cggggtagaa agcctgggag tgtggtggca
     4381 gctgctgctg cagaggccaa aaagaaagct gtgaaggaat cttctatacg gtctgtgcag
     4441 gagactgtgc tccccatcaa gaagcgcaag acccggagcg gcccgggatc cccgaagaag
     4501 aagcgcaaag tatacaccat ggtgagcaag ggcgaggagg ataacatggc catcatcaag
     4561 gagttcatgc gcttcaaggt gcacatggag ggctccgtga acggccacga gttcgagatc
     4621 gagggcgagg gcgagggccg cccctacgag ggcacccaga ccgccaagct gaaggtgacc
     4681 aagggtggcc ccctgccctt cgcctgggac atcctgtccc ctcagttcat gtacggctcc
     4741 aaggcctacg tgaagcaccc cgccgacatc cccgactact tgaagctgtc cttccccgag
     4801 ggcttcaagt gggagcgcgt gatgaacttc gaggacggcg gcgtggtgac cgtgacccag
     4861 gactcctccc tgcaggacgg cgagttcatc tacaaggtga agctgcgcgg caccaacttc
     4921 ccctccgacg gccccgtaat gcagaagaag accatgggct gggaggcctc ctccgagcgg
     4981 atgtaccccg aggacggcgc cctgaagggc gagatcaagc agaggctgaa gctgaaggac
     5041 ggcggccact acgacgctga ggtcaagacc acctacaagg ccaagaagcc cgtgcagctg
     5101 cccggcgcct acaacgtcaa catcaagttg gacatcacct cccacaacga ggactacacc
     5161 atcgtggaac agtacgaacg cgccgagggc cgccactcca ccggcggcat ggacgagctg
     5221 tacaagtaaa gcggccgcga ctctagagcc tgcagtctcg aggcatgcgg taccaagctt
     5281 gtcgagaagt actagaggat cataatcagc cataccacat ttgtagaggt tttacttgct
     5341 ttaaaaaacc tcccacacct ccccctgaac ctgaaacata aaatgaatgc aattgttgtt
     5401 gttaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc
     5461 acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta
     5521 tcttatcatg tctggatctg atcactgctt gagcctagga gatccgaacc agataagtga
     5581 aatctagttc caaactattt tgtcattttt aattttcgta ttagcttacg acgctacacc
     5641 cagttcccat ctattttgtc actcttccct aaataatcct taaaaactcc atttccaccc
     5701 ctcccagttc ccaactattt tgtccgccca cagcggggca tttttcttcc tgttatgttt
     5761 ttaatcaaac atcctgccaa ctccatgtga caaaccgtca tcttcggcta ctttttctct
     5821 gtcacagaat gaaaattttt ctgtcatctc ttcgttatta atgtttgtaa ttgactgaat
     5881 atcaacgctt atttgcagcc tga
//
