LOCUS       sequence-160191-        7581 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of GFP tagged MeCP2-WT.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7581)
  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 7581)
  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
FEATURES             Location/Qualifiers
     source          1..7581
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             1096..1116
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=7xHis
                     /translation="HHHHHHH"
     protein_bind    complement(1457..1481)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             join(1492..1494,1495..1497,1498..2211)
                     /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(1525..1553)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     2148..2168
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     primer_bind     complement(2233..2250)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2239..2463
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2509..2937
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2596..2615)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2806..2827
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2946..2966)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2951..3280
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3131..3266
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3193..3212
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3347..4141
                     /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(3401..3420)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4011..4030
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4317..4438
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4354..4373)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4408..4427
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4487..4503)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4487..4503)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4500..4522)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4511..4527
                     /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(4535..4541,4542..4559,4560..4565))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4580..4601
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4718..4735)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4889..5477)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4969..4988)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5648..6439,6440..6508))
                     /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     6271..6290
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6509..6613)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(6688..6707)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        6879..7258
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        7259..7462
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     7413..7433
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     7507..7526
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        7507..7525
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    join(7558..7581,1)
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
ORIGIN
        1 atggtagctg ggatgttagg gctcagggaa gaaaagtcag aagaccagga cctccagggc
       61 ctcaaggaca aacccctcaa gtttaaaaag gtgaagaaag ataagaaaga agagaaagag
      121 ggcaagcatg agcccgtgca gccatcagcc caccactctg ctgagcccgc agaggcaggc
      181 aaagcagaga catcagaagg gtcaggctcc gccccggctg tgccggaagc ttctgcctcc
      241 cccaaacagc ggcgctccat catccgtgac cggggaccca tgtatgatga ccccaccctg
      301 cctgaaggct ggacacggaa gcttaagcaa aggaaatctg gccgctctgc tgggaagtat
      361 gatgtgtatt tgatcaatcc ccagggaaaa gcctttcgct ctaaagtgga gttgattgcg
      421 tacttcgaaa aggtaggcga cacatccctg gaccctaatg attttgactt cacggtaact
      481 gggagaggga gcccctcccg gcgagagcag aaaccaccta agaagcccaa atctcccaaa
      541 gctccaggaa ctggcagagg ccggggacgc cccaaaggga gcggcaccac gagacccaag
      601 gcggccacgt cagagggtgt gcaggtgaaa agggtcctgg agaaaagtcc tgggaagctc
      661 cttgtcaaga tgccttttca aacttcgcca gggggcaagg ctgagggggg tggggccacc
      721 acatccaccc aggtcatggt gatcaaacgc cccggcagga agcgaaaagc tgaggccgac
      781 cctcaggcca ttcccaagaa acggggccga aagccgggga gtgtggtggc agccgctgcc
      841 gccgaggcca aaaagaaagc cgtgaaggag tcttctatcc gatctgtgca ggagaccgta
      901 ctccccatca agaagcgcaa gacccgggag acggtcagca tcgaggtcaa ggaagtggtg
      961 aagcccctgc tggtgtccac cctcggtgag aagagcggga aaggactgaa gacctgtaag
     1021 agccctgggc ggaaaagcaa ggagagcagc cccaaggggc gcagcagcag cgcctcctca
     1081 ccccccaaga aggagcacca ccaccatcac caccactcag agtccccaaa ggcccccgtg
     1141 ccactgctcc cacccctgcc cccacctcca cctgagcccg agagctccga ggaccccacc
     1201 agcccccctg agccccagga cttgagcagc agcgtctgca aagaggagaa gatgcccaga
     1261 ggaggctcac tggagagcga cggctgcccc aaggagccag ctaagactca gcccgcggtt
     1321 gccaccgccg ccacggccgc agaaaagtac aaacaccgag gggagggaga gcgcaaagac
     1381 attgtttcat cctccatgcc aaggccaaac agagaggagc ctgtggacag ccggacgccc
     1441 gtgaccgaga gagttagccc aactttcttg tacaaagtgg ttcgatctag aatggctagc
     1501 aaaggagaag aacttttcac tggagttgtc ccaattcttg ttgaattaga tggtgatgtt
     1561 aatgggcaca aattttctgt cagtggagag ggtgaaggtg atgctacata cggaaagctt
     1621 acccttaaat ttatttgcac tactggaaaa ctacctgttc catggccaac acttgtcact
     1681 actttctctt atggtgttca atgcttttcc cgttatccgg atcatatgaa acggcatgac
     1741 tttttcaaga gtgccatgcc cgaaggttat gtacaggaac gcactatatc tttcaaagat
     1801 gacgggaact acaagacgcg tgctgaagtc aagtttgaag gtgataccct tgttaatcgt
     1861 atcgagttaa aaggtattga ttttaaagaa gatggaaaca ttctcggaca caaactcgag
     1921 tacaactata actcacacaa tgtatacatc acggcagaca aacaaaagaa tggaatcaaa
     1981 gctaacttca aaattcgcca caacattgaa gatggatccg ttcaactagc agaccattat
     2041 caacaaaata ctccaattgg cgatggccct gtccttttac cagacaacca ttacctgtcg
     2101 acacaatctg ccctttcgaa agatcccaac gaaaagcgtg accacatggt ccttcttgag
     2161 tttgtaactg ctgctgggat tacacatggc atggatgagc tctacaaata atgaattaaa
     2221 cccgctgatc agcctcgact gtgccttcta gttgccagcc atctgttgtt tgcccctccc
     2281 ccgtgccttc cttgaccctg gaaggtgcca ctcccactgt cctttcctaa taaaatgagg
     2341 aaattgcatc gcattgtctg agtaggtgtc attctattct ggggggtggg gtggggcagg
     2401 acagcaaggg ggaggattgg gaagacaata gcaggcatgc tggggatgcg gtgggctcta
     2461 tggcttctga ggcggaaaga accagctggg gctctagggg gtatccccac gcgccctgta
     2521 gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca
     2581 gcgccctagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg ttcgccggct
     2641 ttccccgtca agctctaaat cgggggctcc ctttagggtt ccgatttagt gctttacggc
     2701 acctcgaccc caaaaaactt gattagggtg atggttcacg tagtgggcca tcgccctgat
     2761 agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga ctcttgttcc
     2821 aaactggaac aacactcaac cctatctcgg tctattcttt tgatttataa gggattttgc
     2881 cgatttcggc ctattggtta aaaaatgagc tgatttaaca aaaatttaac gcgaattaat
     2941 tctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag caggcagaag
     3001 tatgcaaagc atgcatctca attagtcagc aaccaggtgt ggaaagtccc caggctcccc
     3061 agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccatag tcccgcccct
     3121 aactccgccc atcccgcccc taactccgcc cagttccgcc cattctccgc cccatggctg
     3181 actaattttt tttatttatg cagaggccga ggccgcctct gcctctgagc tattccagaa
     3241 gtagtgagga ggcttttttg gaggcctagg cttttgcaaa aagctcccgg gagcttgtat
     3301 atccattttc ggatctgatc aagagacagg atgaggatcg tttcgcatga ttgaacaaga
     3361 tggattgcac gcaggttctc cggccgcttg ggtggagagg ctattcggct atgactgggc
     3421 acaacagaca atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc
     3481 ggttcttttt gtcaagaccg acctgtccgg tgccctgaat gaactgcagg acgaggcagc
     3541 gcggctatcg tggctggcca cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac
     3601 tgaagcggga agggactggc tgctattggg cgaagtgccg gggcaggatc tcctgtcatc
     3661 tcaccttgct cctgccgaga aagtatccat catggctgat gcaatgcggc ggctgcatac
     3721 gcttgatccg gctacctgcc cattcgacca ccaagcgaaa catcgcatcg agcgagcacg
     3781 tactcggatg gaagccggtc ttgtcgatca ggatgatctg gacgaagagc atcaggggct
     3841 cgcgccagcc gaactgttcg ccaggctcaa ggcgcgcatg cccgacggcg aggatctcgt
     3901 cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg
     3961 attcatcgac tgtggccggc tgggtgtggc ggaccgctat caggacatag cgttggctac
     4021 ccgtgatatt gctgaagagc ttggcggcga atgggctgac cgcttcctcg tgctttacgg
     4081 tatcgccgct cccgattcgc agcgcatcgc cttctatcgc cttcttgacg agttcttctg
     4141 agcgggactc tggggttcgc gaaatgaccg accaagcgac gcccaacctg ccatcacgag
     4201 atttcgattc caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg
     4261 ccggctggat gatcctccag cgcggggatc tcatgctgga gttcttcgcc caccccaact
     4321 tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata
     4381 aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttatc
     4441 atgtctgtat accgtcgacc tctagctaga gcttggcgta atcatggtca tagctgtttc
     4501 ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt
     4561 gtaaagcctg gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc
     4621 ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg
     4681 ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct
     4741 cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca
     4801 cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga
     4861 accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc
     4921 acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg
     4981 cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat
     5041 acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt
     5101 atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc
     5161 agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg
     5221 acttatcgcc actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg
     5281 gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg
     5341 gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg
     5401 gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca
     5461 gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga
     5521 acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga
     5581 tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt
     5641 ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt
     5701 catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat
     5761 ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag
     5821 caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct
     5881 ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt
     5941 tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg
     6001 cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca
     6061 aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt
     6121 tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat
     6181 gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac
     6241 cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa
     6301 aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt
     6361 tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt
     6421 tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa
     6481 gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt
     6541 atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa
     6601 taggggttcc gcgcacattt ccccgaaaag tgccacctga cgtcgacgga tcgggagatc
     6661 tcccgatccc ctatggtgca ctctcagtac aatctgctct gatgccgcat agttaagcca
     6721 gtatctgctc cctgcttgtg tgttggaggt cgctgagtag tgcgcgagca aaatttaagc
     6781 tacaacaagg caaggcttga ccgacaattg catgaagaat ctgcttaggg ttaggcgttt
     6841 tgcgctgctt cgcgatgtac gggccagata tacgcgttga cattgattat tgactagtta
     6901 ttaatagtaa tcaattacgg ggtcattagt tcatagccca tatatggagt tccgcgttac
     6961 ataacttacg gtaaatggcc cgcctggctg accgcccaac gacccccgcc cattgacgtc
     7021 aataatgacg tatgttccca tagtaacgcc aatagggact ttccattgac gtcaatgggt
     7081 ggagtattta cggtaaactg cccacttggc agtacatcaa gtgtatcata tgccaagtac
     7141 gccccctatt gacgtcaatg acggtaaatg gcccgcctgg cattatgccc agtacatgac
     7201 cttatgggac tttcctactt ggcagtacat ctacgtatta gtcatcgcta ttaccatggt
     7261 gatgcggttt tggcagtaca tcaatgggcg tggatagcgg tttgactcac ggggatttcc
     7321 aagtctccac cccattgacg tcaatgggag tttgttttgg caccaaaatc aacgggactt
     7381 tccaaaatgt cgtaacaact ccgccccatt gacgcaaatg ggcggtaggc gtgtacggtg
     7441 ggaggtctat ataagcagag ctctctggct aactagagaa cccactgctt actggcttat
     7501 cgaaattaat acgactcact atagggagac ccaagctggc tagttaagct tgatcaaaca
     7561 agtttgtaca aaaaagttgg c
//
