LOCUS       sequence-350801-        5732 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  shRNA ctrl.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5732)
  AUTHORS   Yu H, Su Y, Shin J, Zhong C, Guo JU, Weng YL, Gao F, Geschwind DH, 
            Coppola G, Ming GL, Song H
  TITLE     Tet3 regulates synaptic transmission and homeostatic plasticity via 
            DNA oxidation and repair.
  JOURNAL   Nat Neurosci. 2015 Jun;18(6):836-43. doi: 10.1038/nn.4008. Epub 2015
            Apr 27.
  PUBMED    25915473
REFERENCE   2  (bases 1 to 5732)
  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: "10.1038/nn.4008"; 
            journalName: "Nat Neurosci"; date: "2015-06"; volume: "18"; issue: 
            "6"; pages: "836-43"
COMMENT     Sequence Label: AAV-shRNA-ctrl
FEATURES             Location/Qualifiers
     source          1..5732
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   146..286
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   157..286
                     /label=AAV2 ITR
     polyA_signal    326..802
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     584..603
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     misc_feature    834..1422
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     CDS             940..951
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     1349..1369
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(join(1444..2157,2158..2160,2161..2163))
                     /codon_start=1
                     /product="enhanced YFP"
                     /label=EYFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1489..1510)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     1838..1857
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2097..2118
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     promoter        complement(2703..2951)
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     complement(2761..2780)
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     primer_bind     complement(2931..2951)
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     repeat_region   3151..3280
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     rep_origin      complement(3342..3930)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3422..3441)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4101..4892,4893..4961))
                     /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     4724..4743
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4962..5066)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5134..5152
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5190..5212)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5312..5331
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(join(5348..5732,1..71))
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5485..5506)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5697..5716
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 ggcaagtgta gcggtcacgc tgcgcgtaac caccacaccc gccgcgctta atgcgccgct
       61 acagggcgcg tactatggtt gctttgacgt atgcggtgtg aaataccgca cagatgcgta
      121 aggagaaaat accgcatcag gcgcccctgc aggcagctgc gcgctcgctc gctcactgag
      181 gccgcccggg caaagcccgg gcgtcgggcg acctttggtc gcccggcctc agtgagcgag
      241 cgagcgcgca gagagggagt ggccaactcc atcactaggg gttcctgcgg ccgctcggtc
      301 cgcacgtggt tacctacaaa atcagaagga cagggaaggg agcagtggtt cacgcctgta
      361 atcccagcaa tttgggaggc caaggtgggt agatcacctg agattaggag ttggagacca
      421 gcctggccaa tatggtgaaa ccccgtctct accaaaaaaa caaaaattag ctgagcctgg
      481 tcatgcatgc ctggaatccc aacaactcgg gaggctgagg caggagaatc gcttgaaccc
      541 aggaggcgga gattgcagtg agccaagatt gtgccactgc actccagctt ggttcccaat
      601 agaccccgca ggccctacag gttgtcttcc caacttgccc cttgctccat accacccccc
      661 tccaccccat aatattatag aaggacacct agtcagacaa aatgatgcaa cttaatttta
      721 ttaggacaag gctggtgggc actggagtgg caacttccag ggccaggaga ggcactgggg
      781 aggggtcaca gggatgccac ccgtagatct ctcgagcagc gctcggtatc gatgcgggga
      841 ggcggcccaa agggagatcc gactcgtctg agggcgaagg cgaagacgcg gaagaggccg
      901 cagagccggc agcaggccgc gggaaggaag gtccgctgga ttgagggccg aagggacgta
      961 gcagaaggac gtcccgcgca gaatccaggt ggcaacatag gcgagcagcc aaggaaagga
     1021 cgatgatttc cccgacaaca ccacggaatt gtcagtgccc aacagccgag cccctgtcca
     1081 gcagcgggca aggcaggcgg cgatgagttc cgccgtggca atagggaggg ggaaagcgaa
     1141 agtcccggaa aggagctgac aggtggtggc aatgccccaa ccagtggggg ttgcgtcagc
     1201 aaacacagtg cacaccacgc cacgttgcct gacaacgggc cacaactcct cataaagaga
     1261 cagcaaccag gatttataca aggaggagaa aatgaaagcc atacgggaag caatagcatg
     1321 atacaaaggc attaaagcag cgtatccaca tagcgtaaaa ggagcaacat agttaagaat
     1381 accagtcaat ctttcacaaa ttttgtaatc cagaggttga ttatcgataa gcttgatgat
     1441 atcttacttg tacagctcgt ccatgccgag agtgatcccg gcggcggtca cgaactccag
     1501 caggaccatg tgatcgcgct tctcgttggg gtctttgctc agggcggact ggtagctcag
     1561 gtagtggttg tcgggcagca gcacggggcc gtcgccgatg ggggtgttct gctggtagtg
     1621 gtcggcgagc tgcacgctgc cgtcctcgat gttgtggcgg atcttgaagt tcaccttgat
     1681 gccgttcttc tgcttgtcgg ccatgatata gacgttgtgg ctgttgtagt tgtactccag
     1741 cttgtgcccc aggatgttgc cgtcctcctt gaagtcgatg cccttcagct cgatgcggtt
     1801 caccagggtg tcgccctcga acttcacctc ggcgcgggtc ttgtagttgc cgtcgtcctt
     1861 gaagaagatg gtgcgctcct ggacgtagcc ttcgggcatg gcggacttga agaagtcgtg
     1921 ctgcttcatg tggtcggggt agcgggcgaa gcactgcagg ccgtagccga aggtggtcac
     1981 gagggtgggc cagggcacgg gcagcttgcc ggtggtgcag atgaacttca gggtcagctt
     2041 gccgtaggtg gcatcgccct cgccctcgcc ggacacgctg aacttgtggc cgtttacgtc
     2101 gccgtccagc tcgaccagga tgggcaccac cccggtgaac agctcctcgc ccttgctcac
     2161 catgtcgaca gctagaactc gccgcagacc cggtgaggaa gagagcgaac cgggccttaa
     2221 ccacccttta tatagccgcc tctactgggc ggggattaac catgacatca tcaagtccag
     2281 aattcccaaa gtgctagagg cggggtcttg acagaacatt cagcctgtaa gcgtcttgta
     2341 tactacaact cccagtagca ctagggcgga tgcagctgtt gagtgaaacg cgcgagttat
     2401 cacgaagtta gggcgaaagg aagggtggca ctccctagtg cgtcataagc tagcttgcat
     2461 gcctgagaat ttcagaatgt aatgatacct ttgttagata aggctgttta catctgatag
     2521 tggaccttaa gccgacactt aaatgataaa aacggcaaag aattgcaagt ttgatttgca
     2581 ttggaaaggg tcgctggctt ttgtgttaca cgccccttat ttgtgcttga ttagatgatc
     2641 cctctagaaa aaaaggttca gatgtgcggc gagtcaagct tcactcgccg cacatctgaa
     2701 ccggatcctc gtcctttcca caagatatat aaagccaaga aatcgaaata ctttcaagtt
     2761 acggtaagca tatgatagtc cattttaaaa cataatttta aaactgcaaa ctacccaaga
     2821 aattattact ttctacgtca cgtattttgt actaatatct ttgtgtttac agtcaaatta
     2881 attctaatta tctctctaac agccttgtat cgtatatgca aatatgaagg aatcatggga
     2941 aataggccct cttcctgccc agatctaccg gttcaattgc cgacccctcc ccccaacttc
     3001 tcggggactg tgggcgatgt gcgctctgcc cactgacggg caccggagcc aattcccact
     3061 cctttcaaga cctagaaggt ccattagctg caaagattcc tctctgttta aaactttatc
     3121 catctttgca aagcttacgc gtgcggccgc aggaacccct agtgatggag ttggccactc
     3181 cctctctgcg cgctcgctcg ctcactgagg ccgggcgacc aaaggtcgcc cgacgcccgg
     3241 gcggcctcag tgagcgagcg agcgcgcagc tgcctgcagg acatgtgagc aaaaggccag
     3301 caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc
     3361 cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta
     3421 taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg
     3481 ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc
     3541 tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac
     3601 gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac
     3661 ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg
     3721 aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga
     3781 agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt
     3841 agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag
     3901 cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct
     3961 gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg
     4021 atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat
     4081 gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc
     4141 tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg
     4201 gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct
     4261 ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca
     4321 actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg
     4381 ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg
     4441 tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc
     4501 cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag
     4561 ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg
     4621 ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag
     4681 tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat
     4741 agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg
     4801 atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca
     4861 gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca
     4921 aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat
     4981 tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag
     5041 aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgtctaa
     5101 gaaaccatta ttatcatgac attaacctat aaaaataggc gtatcacgag gccctttcgt
     5161 ctcgcgcgtt tcggtgatga cggtgaaaac ctctgacaca tgcagctccc ggagacggtc
     5221 acagcttgtc tgtaagcgga tgccgggagc agacaagccc gtcagggcgc gtcagcgggt
     5281 gttggcgggt gtcggggctg gcttaactat gcggcatcag agcagattgt actgagagtg
     5341 caccataaaa ttgtaaacgt taatattttg ttaaaattcg cgttaaattt ttgttaaatc
     5401 agctcatttt ttaaccaata gaccgaaatc ggcaaaatcc cttataaatc aaaagaatag
     5461 cccgagatag agttgagtgt tgttccagtt tggaacaaga gtccactatt aaagaacgtg
     5521 gactccaacg tcaaagggcg aaaaaccgtc tatcagggcg atggcccact acgtgaacca
     5581 tcacccaaat caagtttttt ggggtcgagg tgccgtaaag cactaaatcg gaaccctaaa
     5641 gggagccccc gatttagagc ttgacgggga aagccggcga acgtggcgag aaaggaaggg
     5701 aagaaagcga aaggagcggg cgctaaggcg ct
//
