LOCUS       sequence-211343-        6863 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 6863)
  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 6863)
  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..6863
                     /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             3592..3612
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             3619..3639
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     primer_bind     complement(3854..3873)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   4126..4266
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4126..4255
                     /label=AAV2 ITR
     rep_origin      4341..4796
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4428..4447)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4638..4659
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4813..4832)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4932..4954
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4992..5010)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5078..5182
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5183..5251,5252..6043)
                     /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(5401..5420)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6214..6802
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6703..6722
                     /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 ccaccatgat taaaatagct acgagaaagt atctggggaa gcagaacgtg
     1141 tacgacatag gtgtggagcg ggatcacaat tttgcgctga agaatggctt tatcgcatct
     1201 tgcttcgact ccgtggaaat ctccggcgtg gaagatcggt tcaacgcctc cctgggcaca
     1261 taccacgatc tgctgaaaat tatcaaggac aaggacttcc tggacaatga ggaaaacgag
     1321 gacattctgg aagatatcgt gctgaccctg acactgtttg aggacagaga gatgatcgag
     1381 gaacggctga aaacctatgc ccacctgttc gacgacaaag tgatgaagca gctgaagcgg
     1441 cggagataca ccggctgggg caggctgagc cggaagctga tcaacggcat ccgggacaag
     1501 cagtccggca agacaatcct ggatttcctg aagtccgacg gcttcgccaa cagaaacttc
     1561 atgcagctga tccacgacga cagcctgacc tttaaagagg acatccagaa agcccaggtg
     1621 tccggccagg gcgatagcct gcacgagcac attgccaatc tggccggcag ccccgccatt
     1681 aagaagggca tcctgcagac agtgaaggtg gtggacgagc tcgtcaaagt gatgggtcgg
     1741 cacaagccag agaatatcgt tatcgaaatg gcaagggaga accaaaccac ccagaagggc
     1801 cagaagaact ctcgggaacg gatgaaaaga atcgaagagg gaattaagga gctgggatct
     1861 cagatactga aggagcaccc tgtggagaat acacagctcc agaacgagaa actctacctg
     1921 tactacctcc agaacgggcg ggacatgtac gttgaccagg aactcgacat caaccggctg
     1981 tccgattatg acgtggacgc tattgttcca cagtccttcc tcaaagatga ctccattgac
     2041 aacaaggtgc tgaccagatc cgataaggcc cgcggtaagt ctgacaatgt tccatcagaa
     2101 gaggtggtca agaagatgaa gaattactgg cggcagctcc tcaacgccaa actgatcacc
     2161 cagcggaagt ttgacaatct gactaaggca gaaagaggag gtctgagcga actcgacaag
     2221 gccggcttta ttaagaggca actggtcgaa acacgccaga ttaccaaaca cgtggcacaa
     2281 atcctcgact ctaggatgaa cactaagtac gatgagaacg ataagctgat cagggaagtg
     2341 aaagtgataa ctctgaagag caagctggtg tctgacttcc ggaaggactt tcaattctac
     2401 aaagttcgcg aaataaacaa ttaccatcat gctcacgatg cctatctcaa tgctgtcgtt
     2461 ggcaccgccc tgatcaagaa ataccctaaa ctggagtctg agttcgtgta cggtgactat
     2521 aaagtctacg atgtgaggaa gatgatagca aagtctgagc aagagattgg caaagccacc
     2581 gccaagtact tcttctactc taatatcatg aatttcttta agactgagat aaccctggct
     2641 aacggcgaaa tccggaagcg cccactgatc gaaacaaacg gagaaacagg agaaatcgtg
     2701 tgggataaag gcagggactt cgcaactgtg cggaaggtgc tgtccatgcc acaagtcaat
     2761 atcgtgaaga agaccgaagt gcagaccggc ggattctcaa aggagagcat cctgccaaag
     2821 cggaactctg acaagctgat cgccaggaag aaagattggg acccaaagaa gtatggcggt
     2881 ttcgattccc ctacagtggc ttattccgtt ctggtcgtgg caaaagtgga gaaaggcaag
     2941 tccaagaaac tcaagtctgt taaggagctg ctcggaatta ctattatgga gagatccagc
     3001 ttcgagaaga atccaatcga tttcctggaa gctaagggct ataaagaagt gaagaaagat
     3061 ctcatcatca aactgcccaa gtactctctc tttgagctgg agaatggtag gaagcggatg
     3121 ctggcctccg ccggagagct gcagaaagga aacgagctgg ctctgccctc caaatacgtg
     3181 aacttcctgt atctggcctc ccactacgag aaactcaaag gtagccctga agacaatgag
     3241 cagaagcaac tctttgttga gcaacataaa cactacctgg acgaaatcat tgaacagatt
     3301 agcgagttca gcaagcgggt tattctggcc gatgcaaacc tcgataaagt gctgagcgca
     3361 tataataagc acagggacaa gccaattcgc gaacaagcag agaatattat ccacctcttt
     3421 actctgacta atctgggcgc tcctgctgcc ttcaagtatt tcgatacaac tattgacagg
     3481 aagcggtaca cctctaccaa agaagttctc gatgccaccc tgatacacca gtcaattacc
     3541 ggactgtacg agactcgcat cgacctgtct cagctcggcg gcgacggttc tcccaagaag
     3601 aagaggaaag tcgaggaccc gaagaaaaag cgcaaggtcg actaagaatt ctacgggtgg
     3661 catccctgtg acccctcccc agtgcctctc ctggccctgg aagttgccac tccagtgccc
     3721 accagccttg tcctaataaa attaagttgc atcattttgt ctgactaggt gtccttctat
     3781 aatattatgg ggtggagggg ggtggtatgg agcaaggggc aagttgggaa gacaacctgt
     3841 agggcctgcg gggtctattg ggaaccaagc tggagtgcag tggcacaatc ttggctcact
     3901 gcaatctccg cctcctgggt tcaagcgatt ctcctgcctc agcctcccga gttgttggga
     3961 ttccaggcat gcatgaccag gctcagctaa tttttgtttt tttggtagag acggggtttc
     4021 accatattgg ccaggctggt ctccaactcc taatctcagg tgatctaccc accttggcct
     4081 cccaaattgc tgggattaca ggcgtgaacc actgctcgcg gccgcaggaa cccctagtga
     4141 tggagttggc cactccctct ctgcgcgctc gctcgctcac tgaggccggg cgaccaaagg
     4201 tcgcccgacg cccgggcttt gcccgggcgg cctcagtgag cgagcgagcg cgcagctgcc
     4261 tgcaggggcg cctgatgcgg tattttctcc ttacgcatct gtgcggtatt tcacaccgca
     4321 tacgtcaaag caaccatagt acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt
     4381 ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt
     4441 cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcgggggct
     4501 ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgatttggg
     4561 tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga
     4621 gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca accctatctc
     4681 gggctattct tttgatttat aagggatttt gccgatttcg gcctattggt taaaaaatga
     4741 gctgatttaa caaaaattta acgcgaattt taacaaaata ttaacgttta caattttatg
     4801 gtgcactctc agtacaatct gctctgatgc cgcatagtta agccagcccc gacacccgcc
     4861 aacacccgct gacgcgccct gacgggcttg tctgctcccg gcatccgctt acagacaagc
     4921 tgtgaccgtc tccgggagct gcatgtgtca gaggttttca ccgtcatcac cgaaacgcgc
     4981 gagacgaaag ggcctcgtga tacgcctatt tttataggtt aatgtcatga taataatggt
     5041 ttcttagacg tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt
     5101 tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca
     5161 ataatattga aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc ttattccctt
     5221 ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctggtga aagtaaaaga
     5281 tgctgaagat cagttgggtg cacgagtggg ttacatcgaa ctggatctca acagcggtaa
     5341 gatccttgag agttttcgcc ccgaagaacg ttttccaatg atgagcactt ttaaagttct
     5401 gctatgtggc gcggtattat cccgtattga cgccgggcaa gagcaactcg gtcgccgcat
     5461 acactattct cagaatgact tggttgagta ctcaccagtc acagaaaagc atcttacgga
     5521 tggcatgaca gtaagagaat tatgcagtgc tgccataacc atgagtgata acactgcggc
     5581 caacttactt ctgacaacga tcggaggacc gaaggagcta accgcttttt tgcacaacat
     5641 gggggatcat gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa
     5701 cgacgagcgt gacaccacga tgcctgtagc aatggcaaca acgttgcgca aactattaac
     5761 tggcgaacta cttactctag cttcccggca acaattaata gactggatgg aggcggataa
     5821 agttgcagga ccacttctgc gctcggccct tccggctggc tggtttattg ctgataaatc
     5881 tggagccggt gagcgtggaa gccgcggtat cattgcagca ctggggccag atggtaagcc
     5941 ctcccgtatc gtagttatct acacgacggg gagtcaggca actatggatg aacgaaatag
     6001 acagatcgct gagataggtg cctcactgat taagcattgg taactgtcag accaagttta
     6061 ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa
     6121 gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc
     6181 gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat
     6241 ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga
     6301 gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt
     6361 ccttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata
     6421 cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac
     6481 cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg
     6541 ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg
     6601 tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag
     6661 cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct
     6721 ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc
     6781 aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt
     6841 ttgctggcct tttgctcaca tgt
//
