LOCUS       sequence-211341-        7274 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 7274)
  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 7274)
  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..7274
                     /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     955..977
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1039..1058
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     CDS             1099..1119
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     primer_bind     complement(4265..4284)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   4537..4677
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4537..4666
                     /label=AAV2 ITR
     rep_origin      4752..5207
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4839..4858)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5049..5070
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5224..5243)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5343..5365
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5403..5421)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5489..5593
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5594..5662,5663..6454)
                     /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(5812..5831)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6625..7213
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7114..7133
                     /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 cgctgcgacg ctgccttcgc cccgtgcccc gctccgccgc
      721 cgcctcgcgc cgcccgcccc ggctctgact gaccgcgtta ctcccacagg tgagcgggcg
      781 ggacggccct tctcctccgg gctgtaatta gcgcttggtt taatgacggc ttgtttcttt
      841 tctgtggctg cgtgaaagcc ttgaggggct ccgggagggc cctttgtgcg gggggagcgg
      901 ctcggggctg tccgcggggg gacggctgcc ttcggggggg acggggcagg gcggggttcg
      961 gcttctggcg tgtgaccggc ggctctagag cctctgctaa ccatgttcat gccttcttct
     1021 ttttcctaca gctcctgggc aacgtgctgg ttattgtgct gtctcatcat tttggcaaag
     1081 aattaccggt ccaccatgcc gaagaaaaag cgcaaggtcg gttctccagc tgagaagagg
     1141 aagcccatcc gggtgctgtc tctctttgat ggaatcgcta cagggctcct ggtgctgaag
     1201 gacttgggca ttcaggtgga ccgctacatt gcctcggagg tgtgtgagga ctccatcacg
     1261 gtgggcatgg tgcggcacca ggggaagatc atgtacgtcg gggacgtccg cagcgtcaca
     1321 cagaagcata tccaggagtg gggcccattc gatctggtga ttgggggcag tccctgcaat
     1381 gacctctcca tcgtcaaccc tgctcgcaag ggcctctacg agggcactgg ccggctcttc
     1441 tttgagttct accgcctcct gcatgatgcg cggcccaagg agggagatga tcgccccttc
     1501 ttctggctct ttgagaatgt ggtggccatg ggcgttagtg acaagaggga catctcgcga
     1561 tttctcgagt ccaaccctgt gatgattgat gccaaagaag tgtcagctgc acacagggcc
     1621 cgctacttct ggggtaacct tcccggtatg aacaggccgt tggcatccac tgtgaatgat
     1681 aagctggagc tgcaggagtg tctggagcat ggcaggatag ccaagttcag caaagtgagg
     1741 accattacta cgaggtcaaa ctccataaag cagggcaaag accagcattt tcctgtcttc
     1801 atgaatgaga aagaggacat cttatggtgc actgaaatgg aaagggtatt tggtttccca
     1861 gtccactata ctgacgtctc caacatgagc cgcttggcga ggcagagact gctgggccgg
     1921 tcatggagcg tgccagtcat ccgccacctc ttcgctccgc tgaaggagta ttttgcgtgt
     1981 gtgagcggca gcgagacccc cggtaccagc gagagcgcca cccccgagag cgacaagaaa
     2041 tactctattg gactggctat cgggacaaac tccgttggct gggccgtcat aaccgacgag
     2101 tataaggtgc caagcaagaa attcaaggtg ctgggtaata ctgaccgcca ttcaatcaag
     2161 aagaacctga tcggagcact cctcttcgac tccggtgaaa ccgctgaagc tactcggctg
     2221 aagcggaccg caaggcggag atacacccgc cgcaagaatc ggatatgtta tctgcaagag
     2281 atcttcagca acgagatggc caaggtggac gacagcttct tccacagact ggaagagtcc
     2341 ttcctggtgg aagaggataa gaagcacgag cggcacccca tcttcggcaa catcgtggac
     2401 gaggtggcct accacgagaa gtaccccacc atctaccacc tgagaaagaa actggtggac
     2461 agcaccgaca aggccgacct gcggctgatc tatctggccc tggcccacat gatcaagttc
     2521 cggggccact tcctgatcga gggcgacctg aaccccgaca acagcgacgt ggacaagctg
     2581 ttcatccagc tggtgcagac ctacaaccag ctgttcgagg aaaaccccat caacgccagc
     2641 ggcgtggacg ccaaggccat cctgtctgcc agactgagca agagcagacg gctggaaaat
     2701 ctgatcgccc agctgcccgg cgagaagaag aatggcctgt tcggaaacct gattgccctg
     2761 agcctgggcc tgacccccaa cttcaagagc aacttcgacc tggccgagga tgccaaactg
     2821 cagctgagca aggacaccta cgacgacgac ctggacaacc tgctggccca gatcggcgac
     2881 cagtacgccg acctgtttct ggccgccaag aacctgtccg acgccatcct gctgagcgac
     2941 atcctgagag tgaacaccga gatcaccaag gcccccctga gcgcctctat gatcaagaga
     3001 tacgacgagc accaccagga cctgaccctg ctgaaagctc tcgtgcggca gcagctgcct
     3061 gagaagtaca aagagatttt cttcgaccag agcaagaacg gctacgccgg ctacattgac
     3121 ggcggagcca gccaggaaga gttctacaag ttcatcaagc ccatcctgga aaagatggac
     3181 ggcaccgagg aactgctcgt gaagctgaac agagaggacc tgctgcggaa gcagcggacc
     3241 ttcgacaacg gcagcatccc ccaccagatc cacctgggag agctgcacgc cattctgcgg
     3301 cggcaggaag atttttaccc attcctgaag gacaaccggg aaaagatcga gaagatcctg
     3361 accttccgca tcccctacta cgtgggccct ctggccaggg gaaacagcag attcgcctgg
     3421 atgaccagaa agagcgagga aaccatcacc ccctggaact tcgaggaagt ggtggacaag
     3481 ggcgcttccg cccagagctt catcgagcgg atgaccaact tcgataagaa cctgcccaac
     3541 gagaaggtgc tgcccaagca cagcctgctg tacgagtact tcaccgtgta taacgagctg
     3601 accaaagtga aatacgtgac cgagggaatg agaaagcccg ccttcctgag cggcgagcag
     3661 aaaaaggcca tcgtggacct gctgttcaag accaaccgga aagtgaccgt gaagcagctg
     3721 aaagaggact acttcaagaa aatcgagtgc ctgtcttatg agacagagat cctcacggtt
     3781 gaatatggtt tgctcccgat cggaaagatt gtagagaaga ggatcgagtg tactgtctat
     3841 tccgttgata acaacggtaa catctatact cagcctgttg ctcaatggca tgacaggggc
     3901 gagcaggaag tattcgaata ctgcctcgaa gatgggagtc tgatccgggc gacaaaggac
     3961 cataagttta tgacagttga cggccaaatg ctccccatcg acgagatatt tgaaagagaa
     4021 cttgatttga tgcgagttga caacctgcct aattaagaat tctacgggtg gcatccctgt
     4081 gacccctccc cagtgcctct cctggccctg gaagttgcca ctccagtgcc caccagcctt
     4141 gtcctaataa aattaagttg catcattttg tctgactagg tgtccttcta taatattatg
     4201 gggtggaggg gggtggtatg gagcaagggg caagttggga agacaacctg tagggcctgc
     4261 ggggtctatt gggaaccaag ctggagtgca gtggcacaat cttggctcac tgcaatctcc
     4321 gcctcctggg ttcaagcgat tctcctgcct cagcctcccg agttgttggg attccaggca
     4381 tgcatgacca ggctcagcta atttttgttt ttttggtaga gacggggttt caccatattg
     4441 gccaggctgg tctccaactc ctaatctcag gtgatctacc caccttggcc tcccaaattg
     4501 ctgggattac aggcgtgaac cactgctcgc ggccgcagga acccctagtg atggagttgg
     4561 ccactccctc tctgcgcgct cgctcgctca ctgaggccgg gcgaccaaag gtcgcccgac
     4621 gcccgggctt tgcccgggcg gcctcagtga gcgagcgagc gcgcagctgc ctgcaggggc
     4681 gcctgatgcg gtattttctc cttacgcatc tgtgcggtat ttcacaccgc atacgtcaaa
     4741 gcaaccatag tacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg
     4801 cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc
     4861 ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg
     4921 gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgatttgg gtgatggttc
     4981 acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt
     5041 ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct cgggctattc
     5101 ttttgattta taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta
     5161 acaaaaattt aacgcgaatt ttaacaaaat attaacgttt acaattttat ggtgcactct
     5221 cagtacaatc tgctctgatg ccgcatagtt aagccagccc cgacacccgc caacacccgc
     5281 tgacgcgccc tgacgggctt gtctgctccc ggcatccgct tacagacaag ctgtgaccgt
     5341 ctccgggagc tgcatgtgtc agaggttttc accgtcatca ccgaaacgcg cgagacgaaa
     5401 gggcctcgtg atacgcctat ttttataggt taatgtcatg ataataatgg tttcttagac
     5461 gtcaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat
     5521 acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg
     5581 aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc cttattccct tttttgcggc
     5641 attttgcctt cctgtttttg ctcacccaga aacgctggtg aaagtaaaag atgctgaaga
     5701 tcagttgggt gcacgagtgg gttacatcga actggatctc aacagcggta agatccttga
     5761 gagttttcgc cccgaagaac gttttccaat gatgagcact tttaaagttc tgctatgtgg
     5821 cgcggtatta tcccgtattg acgccgggca agagcaactc ggtcgccgca tacactattc
     5881 tcagaatgac ttggttgagt actcaccagt cacagaaaag catcttacgg atggcatgac
     5941 agtaagagaa ttatgcagtg ctgccataac catgagtgat aacactgcgg ccaacttact
     6001 tctgacaacg atcggaggac cgaaggagct aaccgctttt ttgcacaaca tgggggatca
     6061 tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa gccataccaa acgacgagcg
     6121 tgacaccacg atgcctgtag caatggcaac aacgttgcgc aaactattaa ctggcgaact
     6181 acttactcta gcttcccggc aacaattaat agactggatg gaggcggata aagttgcagg
     6241 accacttctg cgctcggccc ttccggctgg ctggtttatt gctgataaat ctggagccgg
     6301 tgagcgtgga agccgcggta tcattgcagc actggggcca gatggtaagc cctcccgtat
     6361 cgtagttatc tacacgacgg ggagtcaggc aactatggat gaacgaaata gacagatcgc
     6421 tgagataggt gcctcactga ttaagcattg gtaactgtca gaccaagttt actcatatat
     6481 actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt
     6541 tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc
     6601 cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt
     6661 gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac
     6721 tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg tccttctagt
     6781 gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct
     6841 gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga
     6901 ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac
     6961 acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg
     7021 agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt
     7081 cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc
     7141 tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg
     7201 gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc
     7261 ttttgctcac atgt
//
