LOCUS       sequence-217572-        7268 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Targeted DNA methylation.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7268)
  AUTHORS   Lu Z, Liu Z, Mao W, Wang X, Zheng X, Chen S, Cao B, Huang S, Zhang 
            X, Zhou T, Zhang Y, Huang X, Sun Q, Li JD
  TITLE     Locus-specific DNA methylation of Mecp2 promoter leads to 
            autism-like phenotypes in mice.
  JOURNAL   Cell Death Dis. 2020 Feb 3;11(2):85. doi: 10.1038/s41419-020-2290-x.
  PUBMED    32015323
REFERENCE   2  (bases 1 to 7268)
  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: "Cell Death 
            Dis."; date: "2020-02-3"; volume: "11(2):85. doi"; pages: " 
            10.1038/s41419-020-2290-x"
COMMENT     Sequence Label: pAAV-CAG-NLS-DNMT3A-L-N-spC9-N-intein-hGH
FEATURES             Location/Qualifiers
     source          1..7268
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     78..97
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     CDS             138..158
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     primer_bind     complement(3304..3323)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   3576..3716
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3576..3705
                     /label=AAV2 ITR
     rep_origin      3791..4246
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3878..3897)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4088..4109
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4263..4282)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4382..4404
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4442..4460)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4528..4632
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4633..4701,4702..5493)
                     /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(4851..4870)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5664..6252
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6153..6172
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   6314..6443
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     primer_bind     join(7262..7268,1..16)
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
ORIGIN
        1 cttctggcgt gtgaccggcg gctctagagc ctctgctaac catgttcatg ccttcttctt
       61 tttcctacag ctcctgggca acgtgctggt tattgtgctg tctcatcatt ttggcaaaga
      121 attaccggtc caccatgccg aagaaaaagc gcaaggtcgg ttctccagct gagaagagga
      181 agcccatccg ggtgctgtct ctctttgatg gaatcgctac agggctcctg gtgctgaagg
      241 acttgggcat tcaggtggac cgctacattg cctcggaggt gtgtgaggac tccatcacgg
      301 tgggcatggt gcggcaccag gggaagatca tgtacgtcgg ggacgtccgc agcgtcacac
      361 agaagcatat ccaggagtgg ggcccattcg atctggtgat tgggggcagt ccctgcaatg
      421 acctctccat cgtcaaccct gctcgcaagg gcctctacga gggcactggc cggctcttct
      481 ttgagttcta ccgcctcctg catgatgcgc ggcccaagga gggagatgat cgccccttct
      541 tctggctctt tgagaatgtg gtggccatgg gcgttagtga caagagggac atctcgcgat
      601 ttctcgagtc caaccctgtg atgattgatg ccaaagaagt gtcagctgca cacagggccc
      661 gctacttctg gggtaacctt cccggtatga acaggccgtt ggcatccact gtgaatgata
      721 agctggagct gcaggagtgt ctggagcatg gcaggatagc caagttcagc aaagtgagga
      781 ccattactac gaggtcaaac tccataaagc agggcaaaga ccagcatttt cctgtcttca
      841 tgaatgagaa agaggacatc ttatggtgca ctgaaatgga aagggtattt ggtttcccag
      901 tccactatac tgacgtctcc aacatgagcc gcttggcgag gcagagactg ctgggccggt
      961 catggagcgt gccagtcatc cgccacctct tcgctccgct gaaggagtat tttgcgtgtg
     1021 tgagcggcag cgagaccccc ggtaccagcg agagcgccac ccccgagagc gacaagaaat
     1081 actctattgg actggctatc gggacaaact ccgttggctg ggccgtcata accgacgagt
     1141 ataaggtgcc aagcaagaaa ttcaaggtgc tgggtaatac tgaccgccat tcaatcaaga
     1201 agaacctgat cggagcactc ctcttcgact ccggtgaaac cgctgaagct actcggctga
     1261 agcggaccgc aaggcggaga tacacccgcc gcaagaatcg gatatgttat ctgcaagaga
     1321 tcttcagcaa cgagatggcc aaggtggacg acagcttctt ccacagactg gaagagtcct
     1381 tcctggtgga agaggataag aagcacgagc ggcaccccat cttcggcaac atcgtggacg
     1441 aggtggccta ccacgagaag taccccacca tctaccacct gagaaagaaa ctggtggaca
     1501 gcaccgacaa ggccgacctg cggctgatct atctggccct ggcccacatg atcaagttcc
     1561 ggggccactt cctgatcgag ggcgacctga accccgacaa cagcgacgtg gacaagctgt
     1621 tcatccagct ggtgcagacc tacaaccagc tgttcgagga aaaccccatc aacgccagcg
     1681 gcgtggacgc caaggccatc ctgtctgcca gactgagcaa gagcagacgg ctggaaaatc
     1741 tgatcgccca gctgcccggc gagaagaaga atggcctgtt cggaaacctg attgccctga
     1801 gcctgggcct gacccccaac ttcaagagca acttcgacct ggccgaggat gccaaactgc
     1861 agctgagcaa ggacacctac gacgacgacc tggacaacct gctggcccag atcggcgacc
     1921 agtacgccga cctgtttctg gccgccaaga acctgtccga cgccatcctg ctgagcgaca
     1981 tcctgagagt gaacaccgag atcaccaagg cccccctgag cgcctctatg atcaagagat
     2041 acgacgagca ccaccaggac ctgaccctgc tgaaagctct cgtgcggcag cagctgcctg
     2101 agaagtacaa agagattttc ttcgaccaga gcaagaacgg ctacgccggc tacattgacg
     2161 gcggagccag ccaggaagag ttctacaagt tcatcaagcc catcctggaa aagatggacg
     2221 gcaccgagga actgctcgtg aagctgaaca gagaggacct gctgcggaag cagcggacct
     2281 tcgacaacgg cagcatcccc caccagatcc acctgggaga gctgcacgcc attctgcggc
     2341 ggcaggaaga tttttaccca ttcctgaagg acaaccggga aaagatcgag aagatcctga
     2401 ccttccgcat cccctactac gtgggccctc tggccagggg aaacagcaga ttcgcctgga
     2461 tgaccagaaa gagcgaggaa accatcaccc cctggaactt cgaggaagtg gtggacaagg
     2521 gcgcttccgc ccagagcttc atcgagcgga tgaccaactt cgataagaac ctgcccaacg
     2581 agaaggtgct gcccaagcac agcctgctgt acgagtactt caccgtgtat aacgagctga
     2641 ccaaagtgaa atacgtgacc gagggaatga gaaagcccgc cttcctgagc ggcgagcaga
     2701 aaaaggccat cgtggacctg ctgttcaaga ccaaccggaa agtgaccgtg aagcagctga
     2761 aagaggacta cttcaagaaa atcgagtgcc tgtcttatga gacagagatc ctcacggttg
     2821 aatatggttt gctcccgatc ggaaagattg tagagaagag gatcgagtgt actgtctatt
     2881 ccgttgataa caacggtaac atctatactc agcctgttgc tcaatggcat gacaggggcg
     2941 agcaggaagt attcgaatac tgcctcgaag atgggagtct gatccgggcg acaaaggacc
     3001 ataagtttat gacagttgac ggccaaatgc tccccatcga cgagatattt gaaagagaac
     3061 ttgatttgat gcgagttgac aacctgccta attaagaatt ctacgggtgg catccctgtg
     3121 acccctcccc agtgcctctc ctggccctgg aagttgccac tccagtgccc accagccttg
     3181 tcctaataaa attaagttgc atcattttgt ctgactaggt gtccttctat aatattatgg
     3241 ggtggagggg ggtggtatgg agcaaggggc aagttgggaa gacaacctgt agggcctgcg
     3301 gggtctattg ggaaccaagc tggagtgcag tggcacaatc ttggctcact gcaatctccg
     3361 cctcctgggt tcaagcgatt ctcctgcctc agcctcccga gttgttggga ttccaggcat
     3421 gcatgaccag gctcagctaa tttttgtttt tttggtagag acggggtttc accatattgg
     3481 ccaggctggt ctccaactcc taatctcagg tgatctaccc accttggcct cccaaattgc
     3541 tgggattaca ggcgtgaacc actgctcgcg gccgcaggaa cccctagtga tggagttggc
     3601 cactccctct ctgcgcgctc gctcgctcac tgaggccggg cgaccaaagg tcgcccgacg
     3661 cccgggcttt gcccgggcgg cctcagtgag cgagcgagcg cgcagctgcc tgcaggggcg
     3721 cctgatgcgg tattttctcc ttacgcatct gtgcggtatt tcacaccgca tacgtcaaag
     3781 caaccatagt acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc
     3841 agcgtgaccg ctacacttgc cagcgcctta gcgcccgctc ctttcgcttt cttcccttcc
     3901 tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcgggggct ccctttaggg
     3961 ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgatttggg tgatggttca
     4021 cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc
     4081 tttaatagtg gactcttgtt ccaaactgga acaacactca actctatctc gggctattct
     4141 tttgatttat aagggatttt gccgatttcg gtctattggt taaaaaatga gctgatttaa
     4201 caaaaattta acgcgaattt taacaaaata ttaacgttta caattttatg gtgcactctc
     4261 agtacaatct gctctgatgc cgcatagtta agccagcccc gacacccgcc aacacccgct
     4321 gacgcgccct gacgggcttg tctgctcccg gcatccgctt acagacaagc tgtgaccgtc
     4381 tccgggagct gcatgtgtca gaggttttca ccgtcatcac cgaaacgcgc gagacgaaag
     4441 ggcctcgtga tacgcctatt tttataggtt aatgtcatga taataatggt ttcttagacg
     4501 tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata
     4561 cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga
     4621 aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc ttattccctt ttttgcggca
     4681 ttttgccttc ctgtttttgc tcacccagaa acgctggtga aagtaaaaga tgctgaagat
     4741 cagttgggtg cacgagtggg ttacatcgaa ctggatctca acagcggtaa gatccttgag
     4801 agttttcgcc ccgaagaacg ttttccaatg atgagcactt ttaaagttct gctatgtggc
     4861 gcggtattat cccgtattga cgccgggcaa gagcaactcg gtcgccgcat acactattct
     4921 cagaatgact tggttgagta ctcaccagtc acagaaaagc atcttacgga tggcatgaca
     4981 gtaagagaat tatgcagtgc tgccataacc atgagtgata acactgcggc caacttactt
     5041 ctgacaacga tcggaggacc gaaggagcta accgcttttt tgcacaacat gggggatcat
     5101 gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt
     5161 gacaccacga tgcctgtagc aatggcaaca acgttgcgca aactattaac tggcgaacta
     5221 cttactctag cttcccggca acaattaata gactggatgg aggcggataa agttgcagga
     5281 ccacttctgc gctcggccct tccggctggc tggtttattg ctgataaatc tggagccggt
     5341 gagcgtggaa gccgcggtat cattgcagca ctggggccag atggtaagcc ctcccgtatc
     5401 gtagttatct acacgacggg gagtcaggca actatggatg aacgaaatag acagatcgct
     5461 gagataggtg cctcactgat taagcattgg taactgtcag accaagttta ctcatatata
     5521 ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt
     5581 gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc
     5641 gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg
     5701 caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact
     5761 ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt tcttctagtg
     5821 tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg
     5881 ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac
     5941 tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca
     6001 cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga
     6061 gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc
     6121 ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct
     6181 gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg
     6241 agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct
     6301 tttgctcaca tgtcctgcag gcagctgcgc gctcgctcgc tcactgaggc cgcccgggcg
     6361 tcgggcgacc tttggtcgcc cggcctcagt gagcgagcga gcgcgcagag agggagtggc
     6421 caactccatc actaggggtt cctgcggcct ctagaaagct tcattgacgt caataatgac
     6481 gtatgttccc atagtaacgc caatagggac tttccattga cgtcaatggg tggagtattt
     6541 acggtaaact gcccacttgg cagtacatca agtgtatcat atgccaagta cgccccctat
     6601 tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc cagtacatga ccttatggga
     6661 ctttcctact tggcagtaca tctacgtatt agtcatcgct attaccatgg tcgaggtgag
     6721 ccccacgttc tgcttcactc tccccatctc ccccccctcc ccacccccaa ttttgtattt
     6781 atttattttt taattatttt gtgcagcgat gggggcgggg gggggggnnn nnnnnaggcg
     6841 gggcggggcg gggcgagggg cggggcgggg cgaggcggag aggtgcggcg gcagccaatc
     6901 agagcggcgc gctccgaaag tttcctttta tggcgaggcg gcggcggcgg cggccctata
     6961 aaaagcgaag cgcgcggcgg gcgggagtcg ctgcgcgctg ccttcgcccc gtgccccgct
     7021 ccgccgccgc ctcgcgccgc ccgccccggc tctgactgac cgcgttactc ccacaggtga
     7081 gcgggcggga cggcccttct cctccgggct gtaattagcg cttggtttaa tgacggcttg
     7141 tttcttttct gtggctgcgt gaaagccttg aggggctccg ggagggccct ttgtgcgggg
     7201 ggagcggctc ggggctgtcc gcggggggac ggctgccttc gggggggacg gggcagggcg
     7261 gggttcgg
//
