LOCUS       sequence-265908-        6942 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of GFP tagged MeCP2-G273X.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6942)
  AUTHORS   Baker SA, Chen L, Wilkins AD, Yu P, Lichtarge O, Zoghbi HY
  TITLE     An AT-hook domain in MeCP2 determines the clinical course of Rett 
            syndrome and related disorders.
  JOURNAL   Cell. 2013 Feb 28;152(5):984-96. doi: 10.1016/j.cell.2013.01.038.
  PUBMED    23452848
REFERENCE   2  (bases 1 to 6942)
  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"; doi: 
            "10.1016/j.cell.2013.01"; journalName: "Cell"; date: "2013-02-28- 
            28"; volume: "152"; issue: "5"; pages: "984-96"
COMMENT     Sequence Label: pDEST-hMeCP2-GFP-G273X
FEATURES             Location/Qualifiers
     source          1..6942
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(68..87)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        259..638
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        639..842
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     793..813
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     887..906
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        887..905
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    938..962
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             join(1814..1816,1817..1819,1820..2533)
                     /codon_start=1
                     /product="Cycle 3 GFP (Crameri et al., 1996)"
                     /label=Cycle 3 GFP
                     /note="mammalian codon-optimized"
                     /translation="MASKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFSYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTISFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIK
                     ANFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITHGMDELYK"
     primer_bind     complement(1847..1875)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     2470..2490
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     primer_bind     complement(2555..2572)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2561..2785
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2831..3259
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2918..2937)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3128..3149
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3268..3288)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3273..3602
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3453..3588
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3515..3534
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3669..4463
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(3723..3742)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4333..4352
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4639..4760
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4676..4695)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4730..4749
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4809..4825)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4809..4825)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4822..4844)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4833..4849
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(4857..4863,4864..4881,4882..4887))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4902..4923
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5040..5057)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5211..5799)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5291..5310)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5970..6761,6762..6830))
                     /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     6593..6612
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6831..6935)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ccccgaaaag tgccacctga cgtcgacgga tcgggagatc tcccgatccc ctatggtgca
       61 ctctcagtac aatctgctct gatgccgcat agttaagcca gtatctgctc cctgcttgtg
      121 tgttggaggt cgctgagtag tgcgcgagca aaatttaagc tacaacaagg caaggcttga
      181 ccgacaattg catgaagaat ctgcttaggg ttaggcgttt tgcgctgctt cgcgatgtac
      241 gggccagata tacgcgttga cattgattat tgactagtta ttaatagtaa tcaattacgg
      301 ggtcattagt tcatagccca tatatggagt tccgcgttac ataacttacg gtaaatggcc
      361 cgcctggctg accgcccaac gacccccgcc cattgacgtc aataatgacg tatgttccca
      421 tagtaacgcc aatagggact ttccattgac gtcaatgggt ggagtattta cggtaaactg
      481 cccacttggc agtacatcaa gtgtatcata tgccaagtac gccccctatt gacgtcaatg
      541 acggtaaatg gcccgcctgg cattatgccc agtacatgac cttatgggac tttcctactt
      601 ggcagtacat ctacgtatta gtcatcgcta ttaccatggt gatgcggttt tggcagtaca
      661 tcaatgggcg tggatagcgg tttgactcac ggggatttcc aagtctccac cccattgacg
      721 tcaatgggag tttgttttgg caccaaaatc aacgggactt tccaaaatgt cgtaacaact
      781 ccgccccatt gacgcaaatg ggcggtaggc gtgtacggtg ggaggtctat ataagcagag
      841 ctctctggct aactagagaa cccactgctt actggcttat cgaaattaat acgactcact
      901 atagggagac ccaagctggc tagttaagct tgatcaaaca agtttgtaca aaaaagttgg
      961 catggtagct gggatgttag ggctcaggga agaaaagtca gaagaccagg acctccaggg
     1021 cctcaaggac aaacccctca agtttaaaaa ggtgaagaaa gataagaaag aagagaaaga
     1081 gggcaagcat gagcccgtgc agccatcagc ccaccactct gctgagcccg cagaggcagg
     1141 caaagcagag acatcagaag ggtcaggctc cgccccggct gtgccggaag cttctgcctc
     1201 ccccaaacag cggcgctcca tcatccgtga ccggggaccc atgtatgatg accccaccct
     1261 gcctgaaggc tggacacgga agcttaagca aaggaaatct ggccgctctg ctgggaagta
     1321 tgatgtgtat ttgatcaatc cccagggaaa agcctttcgc tctaaagtgg agttgattgc
     1381 gtacttcgaa aaggtaggcg acacatccct ggaccctaat gattttgact tcacggtaac
     1441 tgggagaggg agcccctccc ggcgagagca gaaaccacct aagaagccca aatctcccaa
     1501 agctccagga actggcagag gccggggacg ccccaaaggg agcggcacca cgagacccaa
     1561 ggcggccacg tcagagggtg tgcaggtgaa aagggtcctg gagaaaagtc ctgggaagct
     1621 ccttgtcaag atgccttttc aaacttcgcc agggggcaag gctgaggggg gtggggccac
     1681 cacatccacc caggtcatgg tgatcaaacg ccccggcagg aagcgaaaag ctgaggccga
     1741 ccctcaggcc attcccaaga aacggggccg aaagccgagt agcagtagca gtagcaaagt
     1801 ggttcgatct agaatggcta gcaaaggaga agaacttttc actggagttg tcccaattct
     1861 tgttgaatta gatggtgatg ttaatgggca caaattttct gtcagtggag agggtgaagg
     1921 tgatgctaca tacggaaagc ttacccttaa atttatttgc actactggaa aactacctgt
     1981 tccatggcca acacttgtca ctactttctc ttatggtgtt caatgctttt cccgttatcc
     2041 ggatcatatg aaacggcatg actttttcaa gagtgccatg cccgaaggtt atgtacagga
     2101 acgcactata tctttcaaag atgacgggaa ctacaagacg cgtgctgaag tcaagtttga
     2161 aggtgatacc cttgttaatc gtatcgagtt aaaaggtatt gattttaaag aagatggaaa
     2221 cattctcgga cacaaactcg agtacaacta taactcacac aatgtataca tcacggcaga
     2281 caaacaaaag aatggaatca aagctaactt caaaattcgc cacaacattg aagatggatc
     2341 cgttcaacta gcagaccatt atcaacaaaa tactccaatt ggcgatggcc ctgtcctttt
     2401 accagacaac cattacctgt cgacacaatc tgccctttcg aaagatccca acgaaaagcg
     2461 tgaccacatg gtccttcttg agtttgtaac tgctgctggg attacacatg gcatggatga
     2521 gctctacaaa taatgaatta aacccgctga tcagcctcga ctgtgccttc tagttgccag
     2581 ccatctgttg tttgcccctc ccccgtgcct tccttgaccc tggaaggtgc cactcccact
     2641 gtcctttcct aataaaatga ggaaattgca tcgcattgtc tgagtaggtg tcattctatt
     2701 ctggggggtg gggtggggca ggacagcaag ggggaggatt gggaagacaa tagcaggcat
     2761 gctggggatg cggtgggctc tatggcttct gaggcggaaa gaaccagctg gggctctagg
     2821 gggtatcccc acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc
     2881 agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc
     2941 tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcgggggct ccctttaggg
     3001 ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgattaggg tgatggttca
     3061 cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc
     3121 tttaatagtg gactcttgtt ccaaactgga acaacactca accctatctc ggtctattct
     3181 tttgatttat aagggatttt gccgatttcg gcctattggt taaaaaatga gctgatttaa
     3241 caaaaattta acgcgaatta attctgtgga atgtgtgtca gttagggtgt ggaaagtccc
     3301 caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccaggt
     3361 gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt
     3421 cagcaaccat agtcccgccc ctaactccgc ccatcccgcc cctaactccg cccagttccg
     3481 cccattctcc gccccatggc tgactaattt tttttattta tgcagaggcc gaggccgcct
     3541 ctgcctctga gctattccag aagtagtgag gaggcttttt tggaggccta ggcttttgca
     3601 aaaagctccc gggagcttgt atatccattt tcggatctga tcaagagaca ggatgaggat
     3661 cgtttcgcat gattgaacaa gatggattgc acgcaggttc tccggccgct tgggtggaga
     3721 ggctattcgg ctatgactgg gcacaacaga caatcggctg ctctgatgcc gccgtgttcc
     3781 ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga
     3841 atgaactgca ggacgaggca gcgcggctat cgtggctggc cacgacgggc gttccttgcg
     3901 cagctgtgct cgacgttgtc actgaagcgg gaagggactg gctgctattg ggcgaagtgc
     3961 cggggcagga tctcctgtca tctcaccttg ctcctgccga gaaagtatcc atcatggctg
     4021 atgcaatgcg gcggctgcat acgcttgatc cggctacctg cccattcgac caccaagcga
     4081 aacatcgcat cgagcgagca cgtactcgga tggaagccgg tcttgtcgat caggatgatc
     4141 tggacgaaga gcatcagggg ctcgcgccag ccgaactgtt cgccaggctc aaggcgcgca
     4201 tgcccgacgg cgaggatctc gtcgtgaccc atggcgatgc ctgcttgccg aatatcatgg
     4261 tggaaaatgg ccgcttttct ggattcatcg actgtggccg gctgggtgtg gcggaccgct
     4321 atcaggacat agcgttggct acccgtgata ttgctgaaga gcttggcggc gaatgggctg
     4381 accgcttcct cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc gccttctatc
     4441 gccttcttga cgagttcttc tgagcgggac tctggggttc gcgaaatgac cgaccaagcg
     4501 acgcccaacc tgccatcacg agatttcgat tccaccgccg ccttctatga aaggttgggc
     4561 ttcggaatcg ttttccggga cgccggctgg atgatcctcc agcgcgggga tctcatgctg
     4621 gagttcttcg cccaccccaa cttgtttatt gcagcttata atggttacaa ataaagcaat
     4681 agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc
     4741 aaactcatca atgtatctta tcatgtctgt ataccgtcga cctctagcta gagcttggcg
     4801 taatcatggt catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac
     4861 atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca
     4921 ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat
     4981 taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc
     5041 tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca
     5101 aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca
     5161 aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg
     5221 ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg
     5281 acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt
     5341 ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt
     5401 tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc
     5461 tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt
     5521 gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt
     5581 agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc
     5641 tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa
     5701 agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt
     5761 tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct
     5821 acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta
     5881 tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa
     5941 agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc
     6001 tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact
     6061 acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc
     6121 tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt
     6181 ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta
     6241 agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg
     6301 tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt
     6361 acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc
     6421 agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt
     6481 actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc
     6541 tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc
     6601 gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa
     6661 ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac
     6721 tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa
     6781 aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt
     6841 tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa
     6901 tgtatttaga aaaataaaca aataggggtt ccgcgcacat tt
//
