LOCUS       sequence-338845-        7336 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AAV vector expressing an HA-tagged TALE-SID4X transcriptional 
            repressor protein targeting the mouse Tet1FL promoter. 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 7336)
  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 7336)
  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(NG)-Tet1FL-NLS-SID4X_2A_EGFP...
FEATURES             Location/Qualifiers
     source          1..7336
                     /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             2976..2996
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             3396..3449
                     /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(3450..3452,3453..4166)
                     /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(3450..3452,3453..4163)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(3492..3513)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3753..3772)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4100..4121
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(4220..4240)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     repeat_region   4599..4739
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4599..4728
                     /label=AAV2 ITR
     rep_origin      4814..5269
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4901..4920)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5111..5132
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5286..5305)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5405..5427
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5465..5483)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5551..5655
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5656..5724,5725..6516)
                     /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(5874..5893)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6687..7275
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7176..7195
                     /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 ttgcgagcaa caacggggga aagcaggcac
     1141 tcgaaaccgt ccagaggttg ctgcctgtgc tgtgccaagc gcacggactt acgccagagc
     1201 aggtcgtggc aattgcgagc catgacgggg gaaagcaggc actcgaaacc gtccagaggt
     1261 tgctgcctgt gctgtgccaa gcgcacggac taaccccaga gcaggtcgtg gcaattgcga
     1321 gccatgacgg gggaaagcag gcactcgaaa ccgtccagag gttgctgcct gtgctgtgcc
     1381 aagcgcacgg gttgacccca gagcaggtcg tggcaattgc gagccatgac gggggaaagc
     1441 aggcactcga aaccgtccag aggttgctgc ctgtgctgtg ccaagcgcac ggcctgaccc
     1501 cagagcaggt cgtggcaatt gcgagccatg acgggggaaa gcaggcactc gaaaccgtcc
     1561 agaggttgct gcctgtgctg tgccaagcgc acggactgac accagagcag gtcgtggcaa
     1621 ttgcgagcaa catcggggga aagcaggcac tcgaaaccgt ccagaggttg ctgcctgtgc
     1681 tgtgccaagc gcacggactt acacccgaac aagtcgtggc aattgcgagc aacaacgggg
     1741 gaaagcaggc actcgaaacc gtccagaggt tgctgcctgt gctgtgccaa gcgcacggac
     1801 ttacgccaga gcaggtcgtg gcaattgcga gccatgacgg gggaaagcag gcactcgaaa
     1861 ccgtccagag gttgctgcct gtgctgtgcc aagcgcacgg actaacccca gagcaggtcg
     1921 tggcaattgc gagcaacgga gggggaaagc aggcactcga aaccgtccag aggttgctgc
     1981 ctgtgctgtg ccaagcgcac gggttgaccc cagagcaggt cgtggcaatt gcgagcaaca
     2041 tcgggggaaa 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 gcgagccatg acgggggaaa gcaggcactc gaaaccgtcc
     2581 agaggttgct gcctgtgctg tgccaagcgc acgggttgac cccagagcag gtcgtggcaa
     2641 ttgcgagcca tgacggggga aagcaggcac tcgaaaccgt ccagaggttg ctgcctgtgc
     2701 tgtgccaagc gcacggactc acgcctgagc aggtagtggc tattgcatcc aatggcgggg
     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 gcagccccaa gaagaagaga aaggtggagg
     3001 ccagcatgaa catccagatg ctgctggagg ccgctgacta cctggaacgg agagagcgcg
     3061 aagccgagca cggatatgct tcaatgctgc ccggaagcgg catgaatatt cagatgctgc
     3121 tggaggctgc tgattacctg gaaaggcgcg aacgggaggc cgaacatggc tatgcttcca
     3181 tgctgcctgg gtctggaatg aatatccaaa tgctgctgga ggcagccgat tacctggaac
     3241 ggagagaaag agaagccgag cacggatacg ccagcatgct gccaggcagc gggatgaaca
     3301 tacaaatgct gctggaggct gccgattacc tggagaggcg cgagagagaa gctgaacatg
     3361 gctatgcctc tatgctgccc ccatggggca gtggagaggg cagaggaagt ctgctaacat
     3421 gcggtgacgt cgaggagaat cctggcccag tgagcaaggg cgaggagctg ttcaccgggg
     3481 tggtgcccat cctggtcgag ctggacggcg acgtaaacgg ccacaagttc agcgtgtccg
     3541 gcgagggcga gggcgatgcc acctacggca agctgaccct gaagttcatc tgcaccaccg
     3601 gcaagctgcc cgtgccctgg cccaccctcg tgaccaccct gacctacggc gtgcagtgct
     3661 tcagccgcta ccccgaccac atgaagcagc acgacttctt caagtccgcc atgcccgaag
     3721 gctacgtcca ggagcgcacc atcttcttca aggacgacgg caactacaag acccgcgccg
     3781 aggtgaagtt cgagggcgac accctggtga accgcatcga gctgaagggc atcgacttca
     3841 aggaggacgg caacatcctg gggcacaagc tggagtacaa ctacaacagc cacaacgtct
     3901 atatcatggc cgacaagcag aagaacggca tcaaggtgaa cttcaagatc cgccacaaca
     3961 tcgaggacgg cagcgtgcag ctcgccgacc actaccagca gaacaccccc atcggcgacg
     4021 gccccgtgct gctgcccgac aaccactacc tgagcaccca gtccgccctg agcaaagacc
     4081 ccaacgagaa gcgcgatcac atggtcctgc tggagttcgt gaccgccgcc gggatcactc
     4141 tcggcatgga cgagctgtac aagtaaaatc aacctctgga ttacaaaatt tgtgaaagat
     4201 tgactggtat tcttaactat gttgctcctt ttacgctatg tggatacgct gctttaatgc
     4261 ctttgtatca tgctattgct tcccgtatgg ctttcatttt ctcctccttg tataaatcct
     4321 ggttagttct tgccacggcg gaactcatcg ccgcctgcct tgcccgctgc tggacagggg
     4381 ctcggctgtt gggcactgac aattccgtgg tgtttatttg tgaaatttgt gatgctattg
     4441 ctttatttgt aaccatctag ctttatttgt gaaatttgtg atgctattgc tttatttgta
     4501 accattataa gctgcaataa acaagttaac aacaacaatt gcattcattt tatgtttcag
     4561 gttcaggggg agatgtggga ggttttttaa gcggccgcag gaacccctag tgatggagtt
     4621 ggccactccc tctctgcgcg ctcgctcgct cactgaggcc gggcgaccaa aggtcgcccg
     4681 acgcccgggc tttgcccggg cggcctcagt gagcgagcga gcgcgcagct gcctgcaggg
     4741 gcgcctgatg cggtattttc tccttacgca tctgtgcggt atttcacacc gcatacgtca
     4801 aagcaaccat agtacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg
     4861 cgcagcgtga ccgctacact tgccagcgcc ttagcgcccg ctcctttcgc tttcttccct
     4921 tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta
     4981 gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgattt gggtgatggt
     5041 tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg
     5101 ttctttaata gtggactctt gttccaaact ggaacaacac tcaactctat ctcgggctat
     5161 tcttttgatt tataagggat tttgccgatt tcggtctatt ggttaaaaaa tgagctgatt
     5221 taacaaaaat ttaacgcgaa ttttaacaaa atattaacgt ttacaatttt atggtgcact
     5281 ctcagtacaa tctgctctga tgccgcatag ttaagccagc cccgacaccc gccaacaccc
     5341 gctgacgcgc cctgacgggc ttgtctgctc ccggcatccg cttacagaca agctgtgacc
     5401 gtctccggga gctgcatgtg tcagaggttt tcaccgtcat caccgaaacg cgcgagacga
     5461 aagggcctcg tgatacgcct atttttatag gttaatgtca tgataataat ggtttcttag
     5521 acgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa
     5581 atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat
     5641 tgaaaaagga agagtatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg
     5701 gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa
     5761 gatcagttgg gtgcacgagt gggttacatc gaactggatc tcaacagcgg taagatcctt
     5821 gagagttttc gccccgaaga acgttttcca atgatgagca cttttaaagt tctgctatgt
     5881 ggcgcggtat tatcccgtat tgacgccggg caagagcaac tcggtcgccg catacactat
     5941 tctcagaatg acttggttga gtactcacca gtcacagaaa agcatcttac ggatggcatg
     6001 acagtaagag aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta
     6061 cttctgacaa cgatcggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat
     6121 catgtaactc gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag
     6181 cgtgacacca cgatgcctgt agcaatggca acaacgttgc gcaaactatt aactggcgaa
     6241 ctacttactc tagcttcccg gcaacaatta atagactgga tggaggcgga taaagttgca
     6301 ggaccacttc tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc
     6361 ggtgagcgtg gaagccgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt
     6421 atcgtagtta tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc
     6481 gctgagatag gtgcctcact gattaagcat tggtaactgt cagaccaagt ttactcatat
     6541 atactttaga ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt
     6601 tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac
     6661 cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc
     6721 ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca
     6781 actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta
     6841 gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct
     6901 ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg
     6961 gactcaagac gatagttacc tgataaggcg cagcggtcgg gctgaacggg gggttcgtgc
     7021 acacagccca gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta
     7081 tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg
     7141 gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt
     7201 cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg
     7261 cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg
     7321 ccttttgctc acatgt
//
