LOCUS       sequence-338847-        6958 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AAV sequence-specific control vector for TALE-Tet1S-SID4X. Synapsin 
            promoter for neuronal expression. C-terminal, self-cleaving EGFP 
            marker..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6958)
  AUTHORS   Greer CB, Wright J, Weiss JD, Lazarenko RM, Moran SP, Zhu J, 
            Chronister KS, Jin AY, Kennedy AJ, Sweatt JD, Kaas GA
  TITLE     Tet1 Isoforms Differentially Regulate Gene Expression, Synaptic 
            Transmission, and Memory in the Mammalian Brain.
  JOURNAL   J Neurosci. 2021 Jan 27;41(4):578-593. doi: 
            10.1523/JNEUROSCI.1821-20.2020. Epub 2020 Dec 1.
  PUBMED    33262245
REFERENCE   2  (bases 1 to 6958)
  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: 
            "10.1523/JNEUROSCI.1821-20.2020"; journalName: "J Neurosci"; date: 
            "2021-01-27- 27"; volume: "41"; issue: "4"; pages: "578-593"
COMMENT     Sequence Label: pAAV_hSyn_HA-TALE(HD)-Tet1S-NLS_2A_EGFP_W3SL
FEATURES             Location/Qualifiers
     source          1..6958
                     /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        181..628
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     CDS             651..677
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             2973..2993
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             3018..3071
                     /codon_start=1
                     /product="2A peptide from Thosea asigna virus capsid 
                     protein"
                     /label=T2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="EGRGSLLTCGDVEENPGP"
     CDS             join(3072..3074,3075..3788)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     CDS             join(3072..3074,3075..3785)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(3114..3135)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3375..3394)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3722..3743
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(3842..3862)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     repeat_region   4221..4361
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4221..4350
                     /label=AAV2 ITR
     rep_origin      4436..4891
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4523..4542)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4733..4754
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4908..4927)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5027..5049
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5087..5105)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5173..5277
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5278..5346,5347..6138)
                     /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(5496..5515)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6309..6897
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6798..6817
                     /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 gcggccgcac gcgttgtaca gtgtctagac tgcagagggc cctgcgtatg
      181 agtgcaagtg ggttttagga ccaggatgag gcggggtggg ggtgcctacc tgacgaccga
      241 ccccgaccca ctggacaagc acccaacccc cattccccaa attgcgcatc ccctatcaga
      301 gagggggagg ggaaacagga tgcggcgagg cgcgtgcgca ctgccagctt cagcaccgcg
      361 gacagtgcct tcgcccccgc ctggcggcgc gcgccaccgc cgcctcagca ctgaaggcgc
      421 gctgacgtca ctcgccggtc ccccgcaaac tccccttccc ggccaccttg gtcgcgtccg
      481 cgccgccgcc ggcccagccg gaccgcacca cgcgaggcgc gagatagggg ggcacgggcg
      541 cgaccatctg cgctgcggcg ccggcgactc agcgctgcct cagtctgcgg tgggcagcgg
      601 aggagtcgtg tcgtgcctga gagcgcagtc gagaaggatc cgccaccatg tacccatacg
      661 atgttccaga ttacgctgta gatttgagaa ctttgggata ttcacagcag cagcaggaaa
      721 agatcaagcc caaagtgagg tcgacagtcg cgcagcatca cgaagcgctg gtgggtcatg
      781 ggtttacaca tgcccacatc gtagccttgt cgcagcaccc tgcagccctt ggcacggtcg
      841 ccgtcaagta ccaggacatg attgcggcgt tgccggaagc cacacatgag gcgatcgtcg
      901 gtgtggggaa acagtggagc ggagcccgag cgcttgaggc cctgttgacg gtcgcgggag
      961 agctgagagg gcctcccctt cagctggaca cgggccagtt gctgaagatc gcgaagcggg
     1021 gaggagtcac ggcggtcgag gcggtgcacg cgtggcgcaa tgcgctcacg ggagcacccc
     1081 tcaacctgac cccagagcag gtcgtggcaa ttgcgagcca tgacggggga aagcaggcac
     1141 tcgaaaccgt ccagaggttg ctgcctgtgc tgtgccaagc gcacggactt acgccagagc
     1201 aggtcgtggc aattgcgagc aacaacgggg gaaagcaggc actcgaaacc gtccagaggt
     1261 tgctgcctgt gctgtgccaa gcgcacggac taaccccaga gcaggtcgtg gcaattgcga
     1321 gccatgacgg gggaaagcag gcactcgaaa ccgtccagag gttgctgcct gtgctgtgcc
     1381 aagcgcacgg gttgacccca gagcaggtcg tggcaattgc gagcaacatc gggggaaagc
     1441 aggcactcga aaccgtccag aggttgctgc ctgtgctgtg ccaagcgcac ggcctgaccc
     1501 cagagcaggt cgtggcaatt gcgagcaaca acgggggaaa gcaggcactc gaaaccgtcc
     1561 agaggttgct gcctgtgctg tgccaagcgc acggactgac accagagcag gtcgtggcaa
     1621 ttgcgagcca tgacggggga aagcaggcac tcgaaaccgt ccagaggttg ctgcctgtgc
     1681 tgtgccaagc gcacggactt acacccgaac aagtcgtggc aattgcgagc catgacgggg
     1741 gaaagcaggc actcgaaacc gtccagaggt tgctgcctgt gctgtgccaa gcgcacggac
     1801 ttacgccaga gcaggtcgtg gcaattgcga gcaacggagg gggaaagcag gcactcgaaa
     1861 ccgtccagag gttgctgcct gtgctgtgcc aagcgcacgg actaacccca gagcaggtcg
     1921 tggcaattgc gagcaacatc gggggaaagc aggcactcga aaccgtccag aggttgctgc
     1981 ctgtgctgtg ccaagcgcac gggttgaccc cagagcaggt cgtggcaatt gcgagcaaca
     2041 acgggggaaa gcaggcactc gaaaccgtcc agaggttgct gcctgtgctg tgccaagcgc
     2101 acggcctgac cccagagcag gtcgtggcaa ttgcgagcca tgacggggga aagcaggcac
     2161 tcgaaaccgt ccagaggttg ctgcctgtgc tgtgccaagc gcacggactg acaccagagc
     2221 aggtcgtggc aattgcgagc aacatcgggg gaaagcaggc actcgaaacc gtccagaggt
     2281 tgctgcctgt gctgtgccaa gcgcacggcc tcaccccaga gcaggtcgtg gcaattgcga
     2341 gccatgacgg gggaaagcag gcactcgaaa ccgtccagag gttgctgcct gtgctgtgcc
     2401 aagcgcacgg acttacgcca gagcaggtcg tggcaattgc gagcaacgga gggggaaagc
     2461 aggcactcga aaccgtccag aggttgctgc ctgtgctgtg ccaagcgcac ggactaaccc
     2521 cagagcaggt cgtggcaatt gcgagcaaca tcgggggaaa gcaggcactc gaaaccgtcc
     2581 agaggttgct gcctgtgctg tgccaagcgc acgggttgac cccagagcag gtcgtggcaa
     2641 ttgcgagcaa cggaggggga aagcaggcac tcgaaaccgt ccagaggttg ctgcctgtgc
     2701 tgtgccaagc gcacggactc acgcctgagc aggtagtggc tattgcatcc catgacgggg
     2761 gcagacccgc actggagtca atcgtggccc agctttcgag gccggacccc gcgctggccg
     2821 cactcactaa tgatcatctt gtagcgctgg cctgcctcgg cggacgaccc gccttggatg
     2881 cggtgaagaa ggggctcccg cacgcgcctg cattgattaa gcggaccaac agaaggattc
     2941 ccgagaggac atcacatcga gtggcagcta gccccaagaa gaagagaaag gtggaggcca
     3001 gctctagagg cagtggagag ggcagaggaa gtctgctaac atgcggtgac gtcgaggaga
     3061 atcctggccc agtgagcaag ggcgaggagc tgttcaccgg ggtggtgccc atcctggtcg
     3121 agctggacgg cgacgtaaac ggccacaagt tcagcgtgtc cggcgagggc gagggcgatg
     3181 ccacctacgg caagctgacc ctgaagttca tctgcaccac cggcaagctg cccgtgccct
     3241 ggcccaccct cgtgaccacc ctgacctacg gcgtgcagtg cttcagccgc taccccgacc
     3301 acatgaagca gcacgacttc ttcaagtccg ccatgcccga aggctacgtc caggagcgca
     3361 ccatcttctt caaggacgac ggcaactaca agacccgcgc cgaggtgaag ttcgagggcg
     3421 acaccctggt gaaccgcatc gagctgaagg gcatcgactt caaggaggac ggcaacatcc
     3481 tggggcacaa gctggagtac aactacaaca gccacaacgt ctatatcatg gccgacaagc
     3541 agaagaacgg catcaaggtg aacttcaaga tccgccacaa catcgaggac ggcagcgtgc
     3601 agctcgccga ccactaccag cagaacaccc ccatcggcga cggccccgtg ctgctgcccg
     3661 acaaccacta cctgagcacc cagtccgccc tgagcaaaga ccccaacgag aagcgcgatc
     3721 acatggtcct gctggagttc gtgaccgccg ccgggatcac tctcggcatg gacgagctgt
     3781 acaagtaaaa tcaacctctg gattacaaaa tttgtgaaag attgactggt attcttaact
     3841 atgttgctcc ttttacgcta tgtggatacg ctgctttaat gcctttgtat catgctattg
     3901 cttcccgtat ggctttcatt ttctcctcct tgtataaatc ctggttagtt cttgccacgg
     3961 cggaactcat cgccgcctgc cttgcccgct gctggacagg ggctcggctg ttgggcactg
     4021 acaattccgt ggtgtttatt tgtgaaattt gtgatgctat tgctttattt gtaaccatct
     4081 agctttattt gtgaaatttg tgatgctatt gctttatttg taaccattat aagctgcaat
     4141 aaacaagtta acaacaacaa ttgcattcat tttatgtttc aggttcaggg ggagatgtgg
     4201 gaggtttttt aagcggccgc aggaacccct agtgatggag ttggccactc cctctctgcg
     4261 cgctcgctcg ctcactgagg ccgggcgacc aaaggtcgcc cgacgcccgg gctttgcccg
     4321 ggcggcctca gtgagcgagc gagcgcgcag ctgcctgcag gggcgcctga tgcggtattt
     4381 tctccttacg catctgtgcg gtatttcaca ccgcatacgt caaagcaacc atagtacgcg
     4441 ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca
     4501 cttgccagcg ccttagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc
     4561 gccggctttc cccgtcaagc tctaaatcgg gggctccctt tagggttccg atttagtgct
     4621 ttacggcacc tcgaccccaa aaaacttgat ttgggtgatg gttcacgtag tgggccatcg
     4681 ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa tagtggactc
     4741 ttgttccaaa ctggaacaac actcaactct atctcgggct attcttttga tttataaggg
     4801 attttgccga tttcggtcta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg
     4861 aattttaaca aaatattaac gtttacaatt ttatggtgca ctctcagtac aatctgctct
     4921 gatgccgcat agttaagcca gccccgacac ccgccaacac ccgctgacgc gccctgacgg
     4981 gcttgtctgc tcccggcatc cgcttacaga caagctgtga ccgtctccgg gagctgcatg
     5041 tgtcagaggt tttcaccgtc atcaccgaaa cgcgcgagac gaaagggcct cgtgatacgc
     5101 ctatttttat aggttaatgt catgataata atggtttctt agacgtcagg tggcactttt
     5161 cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat
     5221 ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtatg
     5281 agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt
     5341 tttgctcacc cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcacga
     5401 gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa
     5461 gaacgttttc caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt
     5521 attgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt
     5581 gagtactcac cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc
     5641 agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac aacgatcgga
     5701 ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat
     5761 cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct
     5821 gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc
     5881 cggcaacaat taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg
     5941 gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg tggaagccgc
     6001 ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg
     6061 acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca
     6121 ctgattaagc attggtaact gtcagaccaa gtttactcat atatacttta gattgattta
     6181 aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc
     6241 aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa
     6301 ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca
     6361 ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta
     6421 actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc
     6481 caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca
     6541 gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta
     6601 ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag
     6661 cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt
     6721 cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc
     6781 acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac
     6841 ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac
     6901 gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgt
//
