LOCUS       sequence-169559-        5735 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  shRNA against mouse Tet2.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5735)
  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 5735)
  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-Tet2
FEATURES             Location/Qualifiers
     source          1..5735
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..249
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     1..21
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     172..191
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     CDS             join(792..794,795..797,798..1511)
                     /codon_start=1
                     /product="enhanced YFP"
                     /label=EYFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(837..858)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1098..1117)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1445..1466
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    1533..2121
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(1586..1606)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2004..2015)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    2153..2629
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(2352..2371)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   2669..2809
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2669..2798
                     /label=AAV2 ITR
     rep_origin      2884..3339
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2971..2990)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3181..3202
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3356..3375)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3475..3497
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3535..3553)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3621..3725
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3726..3794,3795..4586)
                     /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(3944..3963)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4757..5345
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5246..5265
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   5407..5536
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
ORIGIN
        1 gagggcctat ttcccatgat tccttcatat ttgcatatac gatacaaggc tgttagagag
       61 ataattagaa ttaatttgac tgtaaacaca aagatattag tacaaaatac gtgacgtaga
      121 aagtaataat ttcttgggta gtttgcagtt ttaaaattat gttttaaaat ggactatcat
      181 atgcttaccg taacttgaaa gtatttcgat ttcttggctt tatatatctt gtggaaagga
      241 cgaggatccg ggatgtaagt ttgccagaag cgaagcttgg cttctggcaa acttacatcc
      301 cttttttcta gagggatcat ctaatcaagc acaaataagg ggcgtgtaac acaaaagcca
      361 gcgacccttt ccaatgcaaa tcaaacttgc aattctttgc cgtttttatc atttaagtgt
      421 cggcttaagg tccactatca gatgtaaaca gccttatcta acaaaggtat cattacattc
      481 tgaaattctc aggcatgcaa gctagcttat gacgcactag ggagtgccac ccttcctttc
      541 gccctaactt cgtgataact cgcgcgtttc actcaacagc tgcatccgcc ctagtgctac
      601 tgggagttgt agtatacaag acgcttacag gctgaatgtt ctgtcaagac cccgcctcta
      661 gcactttggg aattctggac ttgatgatgt catggttaat ccccgcccag tagaggcggc
      721 tatataaagg gtggttaagg cccggttcgc tctcttcctc accgggtctg cggcgagttc
      781 tagctgtcga catggtgagc aagggcgagg agctgttcac cggggtggtg cccatcctgg
      841 tcgagctgga cggcgacgta aacggccaca agttcagcgt gtccggcgag ggcgagggcg
      901 atgccaccta cggcaagctg accctgaagt tcatctgcac caccggcaag ctgcccgtgc
      961 cctggcccac cctcgtgacc accttcggct acggcctgca gtgcttcgcc cgctaccccg
     1021 accacatgaa gcagcacgac ttcttcaagt ccgccatgcc cgaaggctac gtccaggagc
     1081 gcaccatctt cttcaaggac gacggcaact acaagacccg cgccgaggtg aagttcgagg
     1141 gcgacaccct ggtgaaccgc atcgagctga agggcatcga cttcaaggag gacggcaaca
     1201 tcctggggca caagctggag tacaactaca acagccacaa cgtctatatc atggccgaca
     1261 agcagaagaa cggcatcaag gtgaacttca agatccgcca caacatcgag gacggcagcg
     1321 tgcagctcgc cgaccactac cagcagaaca cccccatcgg cgacggcccc gtgctgctgc
     1381 ccgacaacca ctacctgagc taccagtccg ccctgagcaa agaccccaac gagaagcgcg
     1441 atcacatggt cctgctggag ttcgtgaccg ccgccgggat cactctcggc atggacgagc
     1501 tgtacaagta agatatcatc aagcttatcg ataatcaacc tctggattac aaaatttgtg
     1561 aaagattgac tggtattctt aactatgttg ctccttttac gctatgtgga tacgctgctt
     1621 taatgccttt gtatcatgct attgcttccc gtatggcttt cattttctcc tccttgtata
     1681 aatcctggtt gctgtctctt tatgaggagt tgtggcccgt tgtcaggcaa cgtggcgtgg
     1741 tgtgcactgt gtttgctgac gcaaccccca ctggttgggg cattgccacc acctgtcagc
     1801 tcctttccgg gactttcgct ttccccctcc ctattgccac ggcggaactc atcgccgcct
     1861 gccttgcccg ctgctggaca ggggctcggc tgttgggcac tgacaattcc gtggtgttgt
     1921 cggggaaatc atcgtccttt ccttggctgc tcgcctatgt tgccacctgg attctgcgcg
     1981 ggacgtcctt ctgctacgtc ccttcggccc tcaatccagc ggaccttcct tcccgcggcc
     2041 tgctgccggc tctgcggcct cttccgcgtc ttcgccttcg ccctcagacg agtcggatct
     2101 ccctttgggc cgcctccccg catcgatacc gagcgctgct cgagagatct acgggtggca
     2161 tccctgtgac ccctccccag tgcctctcct ggccctggaa gttgccactc cagtgcccac
     2221 cagccttgtc ctaataaaat taagttgcat cattttgtct gactaggtgt ccttctataa
     2281 tattatgggg tggagggggg tggtatggag caaggggcaa gttgggaaga caacctgtag
     2341 ggcctgcggg gtctattggg aaccaagctg gagtgcagtg gcacaatctt ggctcactgc
     2401 aatctccgcc tcctgggttc aagcgattct cctgcctcag cctcccgagt tgttgggatt
     2461 ccaggcatgc atgaccaggc tcagctaatt tttgtttttt tggtagagac ggggtttcac
     2521 catattggcc aggctggtct ccaactccta atctcaggtg atctacccac cttggcctcc
     2581 caaattgctg ggattacagg cgtgaaccac tgctcccttc cctgtccttc tgattttgta
     2641 ggtaaccacg tgcggaccga gcggccgcag gaacccctag tgatggagtt ggccactccc
     2701 tctctgcgcg ctcgctcgct cactgaggcc gggcgaccaa aggtcgcccg acgcccgggc
     2761 tttgcccggg cggcctcagt gagcgagcga gcgcgcagct gcctgcaggg gcgcctgatg
     2821 cggtattttc tccttacgca tctgtgcggt atttcacacc gcatacgtca aagcaaccat
     2881 agtacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga
     2941 ccgctacact tgccagcgcc ttagcgcccg ctcctttcgc tttcttccct tcctttctcg
     3001 ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat
     3061 ttagtgcttt acggcacctc gaccccaaaa aacttgattt gggtgatggt tcacgtagtg
     3121 ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata
     3181 gtggactctt gttccaaact ggaacaacac tcaactctat ctcgggctat tcttttgatt
     3241 tataagggat tttgccgatt tcggtctatt ggttaaaaaa tgagctgatt taacaaaaat
     3301 ttaacgcgaa ttttaacaaa atattaacgt ttacaatttt atggtgcact ctcagtacaa
     3361 tctgctctga tgccgcatag ttaagccagc cccgacaccc gccaacaccc gctgacgcgc
     3421 cctgacgggc ttgtctgctc ccggcatccg cttacagaca agctgtgacc gtctccggga
     3481 gctgcatgtg tcagaggttt tcaccgtcat caccgaaacg cgcgagacga aagggcctcg
     3541 tgatacgcct atttttatag gttaatgtca tgataataat ggtttcttag acgtcaggtg
     3601 gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa
     3661 atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga
     3721 agagtatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc
     3781 ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg
     3841 gtgcacgagt gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc
     3901 gccccgaaga acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat
     3961 tatcccgtat tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg
     4021 acttggttga gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag
     4081 aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa
     4141 cgatcggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc
     4201 gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca
     4261 cgatgcctgt agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc
     4321 tagcttcccg gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc
     4381 tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg
     4441 ggtctcgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta
     4501 tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag
     4561 gtgcctcact gattaagcat tggtaactgt cagaccaagt ttactcatat atactttaga
     4621 ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc
     4681 tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa
     4741 agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa
     4801 aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc
     4861 cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt
     4921 agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc
     4981 tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac
     5041 gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca
     5101 gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg
     5161 ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag
     5221 gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt
     5281 ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat
     5341 ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc
     5401 acatgtcctg caggcagctg cgcgctcgct cgctcactga ggccgcccgg gcgtcgggcg
     5461 acctttggtc gcccggcctc agtgagcgag cgagcgcgca gagagggagt ggccaactcc
     5521 atcactaggg gttcctgcgg ccgcacgcgt aagctttgca aagatggata aagttttaaa
     5581 cagagaggaa tctttgcagc taatggacct tctaggtctt gaaaggagtg ggaattggct
     5641 ccggtgcccg tcagtgggca gagcgcacat cgcccacagt ccccgagaag ttggggggag
     5701 gggtcggcaa ttgaaccggt agatctgggc aggaa
//
