LOCUS       sequence-217577-        6862 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 6862)
  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 6862)
  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-C-intein-C-spC9-H840A-N863A-2xNLS-hGH
FEATURES             Location/Qualifiers
     source          1..6862
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..130
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     promoter        398..675
                     /label=chicken beta-actin promoter
     primer_bind     954..976
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1038..1057
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     CDS             3591..3611
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             3618..3638
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     primer_bind     complement(3853..3872)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   4125..4265
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4125..4254
                     /label=AAV2 ITR
     rep_origin      4340..4795
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4427..4446)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4637..4658
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4812..4831)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4931..4953
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4991..5009)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5077..5181
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5182..5250,5251..6042)
                     /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(5400..5419)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6213..6801
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6702..6721
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcgtcg ggcgaccttt
       61 ggtcgcccgg cctcagtgag cgagcgagcg cgcagagagg gagtggccaa ctccatcact
      121 aggggttcct gcggcctcta gaaagcttca ttgacgtcaa taatgacgta tgttcccata
      181 gtaacgccaa tagggacttt ccattgacgt caatgggtgg agtatttacg gtaaactgcc
      241 cacttggcag tacatcaagt gtatcatatg ccaagtacgc cccctattga cgtcaatgac
      301 ggtaaatggc ccgcctggca ttatgcccag tacatgacct tatgggactt tcctacttgg
      361 cagtacatct acgtattagt catcgctatt accatggtcg aggtgagccc cacgttctgc
      421 ttcactctcc ccatctcccc cccctcccca cccccaattt tgtatttatt tattttttaa
      481 ttattttgtg cagcgatggg ggcggggggg gggggggggc gcgcgccagg cggggcgggg
      541 cggggcgagg ggcggggcgg ggcgaggcgg agaggtgcgg cggcagccaa tcagagcggc
      601 gcgctccgaa agtttccttt tatggcgagg cggcggcggc ggcggcccta taaaaagcga
      661 agcgcgcggc gggcgggagt cgctgcgcgc tgccttcgcc ccgtgccccg ctccgccgcc
      721 gcctcgcgcc gcccgccccg gctctgactg accgcgttac tcccacaggt gagcgggcgg
      781 gacggccctt ctcctccggg ctgtaattag cgcttggttt aatgacggct tgtttctttt
      841 ctgtggctgc gtgaaagcct tgaggggctc cgggagggcc ctttgtgcgg ggggagcggc
      901 tcggggctgt ccgcgggggg acggctgcct tcggggggga cggggcaggg cggggttcgg
      961 cttctggcgt gtgaccggcg gctctagagc ctctgctaac catgttcatg ccttcttctt
     1021 tttcctacag ctcctgggca acgtgctggt tattgtgctg tctcatcatt ttggcaaaga
     1081 attaccggtc caccatgatt aaaatagcta cgagaaagta tctggggaag cagaacgtgt
     1141 acgacatagg tgtggagcgg gatcacaatt ttgcgctgaa gaatggcttt atcgcatctt
     1201 gcttcgactc cgtggaaatc tccggcgtgg aagatcggtt caacgcctcc ctgggcacat
     1261 accacgatct gctgaaaatt atcaaggaca aggacttcct ggacaatgag gaaaacgagg
     1321 acattctgga agatatcgtg ctgaccctga cactgtttga ggacagagag atgatcgagg
     1381 aacggctgaa aacctatgcc cacctgttcg acgacaaagt gatgaagcag ctgaagcggc
     1441 ggagatacac cggctggggc aggctgagcc ggaagctgat caacggcatc cgggacaagc
     1501 agtccggcaa gacaatcctg gatttcctga agtccgacgg cttcgccaac agaaacttca
     1561 tgcagctgat ccacgacgac agcctgacct ttaaagagga catccagaaa gcccaggtgt
     1621 ccggccaggg cgatagcctg cacgagcaca ttgccaatct ggccggcagc cccgccatta
     1681 agaagggcat cctgcagaca gtgaaggtgg tggacgagct cgtcaaagtg atgggtcggc
     1741 acaagccaga gaatatcgtt atcgaaatgg caagggagaa ccaaaccacc cagaagggcc
     1801 agaagaactc tcgggaacgg atgaaaagaa tcgaagaggg aattaaggag ctgggatctc
     1861 agatactgaa ggagcaccct gtggagaata cacagctcca gaacgagaaa ctctacctgt
     1921 actacctcca gaacgggcgg gacatgtacg ttgaccagga actcgacatc aaccggctgt
     1981 ccgattatga cgtggacgct attgttccac agtccttcct caaagatgac tccattgaca
     2041 acaaggtgct gaccagatcc gataaggccc gcggtaagtc tgacaatgtt ccatcagaag
     2101 aggtggtcaa gaagatgaag aattactggc ggcagctcct caacgccaaa ctgatcaccc
     2161 agcggaagtt tgacaatctg actaaggcag aaagaggagg tctgagcgaa ctcgacaagg
     2221 ccggctttat taagaggcaa ctggtcgaaa cacgccagat taccaaacac gtggcacaaa
     2281 tcctcgactc taggatgaac actaagtacg atgagaacga taagctgatc agggaagtga
     2341 aagtgataac tctgaagagc aagctggtgt ctgacttccg gaaggacttt caattctaca
     2401 aagttcgcga aataaacaat taccatcatg ctcacgatgc ctatctcaat gctgtcgttg
     2461 gcaccgccct gatcaagaaa taccctaaac tggagtctga gttcgtgtac ggtgactata
     2521 aagtctacga tgtgaggaag atgatagcaa agtctgagca agagattggc aaagccaccg
     2581 ccaagtactt cttctactct aatatcatga atttctttaa gactgagata accctggcta
     2641 acggcgaaat ccggaagcgc ccactgatcg aaacaaacgg agaaacagga gaaatcgtgt
     2701 gggataaagg cagggacttc gcaactgtgc ggaaggtgct gtccatgcca caagtcaata
     2761 tcgtgaagaa gaccgaagtg cagaccggcg gattctcaaa ggagagcatc ctgccaaagc
     2821 ggaactctga caagctgatc gccaggaaga aagattggga cccaaagaag tatggcggtt
     2881 tcgattcccc tacagtggct tattccgttc tggtcgtggc aaaagtggag aaaggcaagt
     2941 ccaagaaact caagtctgtt aaggagctgc tcggaattac tattatggag agatccagct
     3001 tcgagaagaa tccaatcgat ttcctggaag ctaagggcta taaagaagtg aagaaagatc
     3061 tcatcatcaa actgcccaag tactctctct ttgagctgga gaatggtagg aagcggatgc
     3121 tggcctccgc cggagagctg cagaaaggaa acgagctggc tctgccctcc aaatacgtga
     3181 acttcctgta tctggcctcc cactacgaga aactcaaagg tagccctgaa gacaatgagc
     3241 agaagcaact ctttgttgag caacataaac actacctgga cgaaatcatt gaacagatta
     3301 gcgagttcag caagcgggtt attctggccg atgcaaacct cgataaagtg ctgagcgcat
     3361 ataataagca cagggacaag ccaattcgcg aacaagcaga gaatattatc cacctcttta
     3421 ctctgactaa tctgggcgct cctgctgcct tcaagtattt cgatacaact attgacagga
     3481 agcggtacac ctctaccaaa gaagttctcg atgccaccct gatacaccag tcaattaccg
     3541 gactgtacga gactcgcatc gacctgtctc agctcggcgg cgacggttct cccaagaaga
     3601 agaggaaagt cgaggacccg aagaaaaagc gcaaggtcga ctaagaattc tacgggtggc
     3661 atccctgtga cccctcccca gtgcctctcc tggccctgga agttgccact ccagtgccca
     3721 ccagccttgt cctaataaaa ttaagttgca tcattttgtc tgactaggtg tccttctata
     3781 atattatggg gtggaggggg gtggtatgga gcaaggggca agttgggaag acaacctgta
     3841 gggcctgcgg ggtctattgg gaaccaagct ggagtgcagt ggcacaatct tggctcactg
     3901 caatctccgc ctcctgggtt caagcgattc tcctgcctca gcctcccgag ttgttgggat
     3961 tccaggcatg catgaccagg ctcagctaat ttttgttttt ttggtagaga cggggtttca
     4021 ccatattggc caggctggtc tccaactcct aatctcaggt gatctaccca ccttggcctc
     4081 ccaaattgct gggattacag gcgtgaacca ctgctcgcgg ccgcaggaac ccctagtgat
     4141 ggagttggcc actccctctc tgcgcgctcg ctcgctcact gaggccgggc gaccaaaggt
     4201 cgcccgacgc ccgggctttg cccgggcggc ctcagtgagc gagcgagcgc gcagctgcct
     4261 gcaggggcgc ctgatgcggt attttctcct tacgcatctg tgcggtattt cacaccgcat
     4321 acgtcaaagc aaccatagta cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg
     4381 gttacgcgca gcgtgaccgc tacacttgcc agcgccttag cgcccgctcc tttcgctttc
     4441 ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcgggggctc
     4501 cctttagggt tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgatttgggt
     4561 gatggttcac gtagtgggcc atcgccctga tagacggttt ttcgcccttt gacgttggag
     4621 tccacgttct ttaatagtgg actcttgttc caaactggaa caacactcaa ctctatctcg
     4681 ggctattctt ttgatttata agggattttg ccgatttcgg tctattggtt aaaaaatgag
     4741 ctgatttaac aaaaatttaa cgcgaatttt aacaaaatat taacgtttac aattttatgg
     4801 tgcactctca gtacaatctg ctctgatgcc gcatagttaa gccagccccg acacccgcca
     4861 acacccgctg acgcgccctg acgggcttgt ctgctcccgg catccgctta cagacaagct
     4921 gtgaccgtct ccgggagctg catgtgtcag aggttttcac cgtcatcacc gaaacgcgcg
     4981 agacgaaagg gcctcgtgat acgcctattt ttataggtta atgtcatgat aataatggtt
     5041 tcttagacgt caggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt
     5101 ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa
     5161 taatattgaa aaaggaagag tatgagtatt caacatttcc gtgtcgccct tattcccttt
     5221 tttgcggcat tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat
     5281 gctgaagatc agttgggtgc acgagtgggt tacatcgaac tggatctcaa cagcggtaag
     5341 atccttgaga gttttcgccc cgaagaacgt tttccaatga tgagcacttt taaagttctg
     5401 ctatgtggcg cggtattatc ccgtattgac gccgggcaag agcaactcgg tcgccgcata
     5461 cactattctc agaatgactt ggttgagtac tcaccagtca cagaaaagca tcttacggat
     5521 ggcatgacag taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc
     5581 aacttacttc tgacaacgat cggaggaccg aaggagctaa ccgctttttt gcacaacatg
     5641 ggggatcatg taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac
     5701 gacgagcgtg acaccacgat gcctgtagca atggcaacaa cgttgcgcaa actattaact
     5761 ggcgaactac ttactctagc ttcccggcaa caattaatag actggatgga ggcggataaa
     5821 gttgcaggac cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct
     5881 ggagccggtg agcgtggaag ccgcggtatc attgcagcac tggggccaga tggtaagccc
     5941 tcccgtatcg tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga
     6001 cagatcgctg agataggtgc ctcactgatt aagcattggt aactgtcaga ccaagtttac
     6061 tcatatatac tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag
     6121 atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg
     6181 tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc
     6241 tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag
     6301 ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtt
     6361 cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac
     6421 ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc
     6481 gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt
     6541 tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt
     6601 gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc
     6661 ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt
     6721 tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca
     6781 ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt
     6841 tgctggcctt ttgctcacat gt
//
