LOCUS       sequence-403393-        6201 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For production of driver AAV virus for Purkinje cell-specific 
            expression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6201)
  AUTHORS   Chen X, Du Y, Broussard GJ, Kislin M, Yuede CM, Zhang S, Dietmann S,
            Gabel H, Zhao G, Wang SS, Zhang X, Bonni A
  TITLE     Transcriptomic mapping uncovers Purkinje neuron plasticity driving 
            learning.
  JOURNAL   Nature. 2022 May;605(7911):722-727. doi: 10.1038/s41586-022-04711-3.
            Epub 2022 May 11.
  PUBMED    35545673
REFERENCE   2  (bases 1 to 6201)
  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/s41586-022-04711-3"; journalName: "Nature"; date: 
            "2022-05"; volume: "605"; issue: "7911"; pages: "722-727"
COMMENT     Sequence Label: pAAV-sL7-Cre-HA-WPRE
FEATURES             Location/Qualifiers
     source          1..6201
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   3..132
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     CDS             1158..1178
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             2205..2294
                     /codon_start=1
                     /product="three tandem HA epitope tags"
                     /label=3xHA
                     /translation="YPYDVPDYAGYPYDVPDYAGSYPYDVPDYA"
     misc_feature    2330..2918
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2383..2403)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2801..2812)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    2950..3426
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(3149..3168)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   3466..3606
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3466..3595
                     /label=AAV2 ITR
     rep_origin      3681..4136
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3768..3787)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3978..3999
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4153..4172)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4272..4294
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4332..4350)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4418..4522
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4523..4591,4592..5383)
                     /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(4741..4760)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5554..6142
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6043..6062
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gtcctgcagg cagctgcgcg ctcgctcgct cactgaggcc gcccgggcgt cgggcgacct
       61 ttggtcgccc ggcctcagtg agcgagcgag cgcgcagaga gggagtggcc aactccatca
      121 ctaggggttc ctgcggccgc acgcgtcatg ttggttgatc ttcaacattg tactgacttc
      181 ttcccacttg acattcctca aggtcctgtg atcatggctg ggtctagtga ggttcaaacc
      241 tgcactgccc tacccacacc cacacccagc tcagcgtcag tcaggatcaa caattaccta
      301 gagatcatct ttctggggct taagcattgg tgggagcaga tgggatatga gctggggatt
      361 tgggaatggg ggaagatatc tgctccccct ccccctacac cctagccttt taaaaggcct
      421 tctcaggtca gagaccagga gaaaagtata ggagagatac acaatggacc aggaagaaga
      481 aaagggagag ggaggctcag accttctaga caaggtaaga gggctctggc tgactccacc
      541 atccgcttct tgaggtctcg gcacctgtaa ttgacaagat taattcattt atagggcatc
      601 taattagcaa gcaagtctct ggagtcccct gacccagtta ctataacaca cagggggtat
      661 aggtaggaga gtataagagc ccctcctcag ggcaaatgaa tggattctta gtactgtccc
      721 ccaagagata gtaggtacta ggatttaggg gcacttctga gccccatttc cctggtaagt
      781 gtcccaaccc cccaaatcaa cccaagcctg gtctcaatct aggacagtgg tagaatgctg
      841 tccctagagt cagtaccatg tgaaattgtg ctgcaggcag gggccccagg ctgggaggtg
      901 ggggttgggg gagtcagggg caggtcaggg aaggagactc aggtttcatt tagagaaatt
      961 ctgcagaccc gtgaggacta tggtgagagc agagatggga aggcaggcac tgtttcgggt
     1021 ggatgctgtc tggaagacag ggaaggcaca gaccaaacta aaccaatcac gtctgtcccc
     1081 aaggcaggtt caccggacca ggaaggcttc ttcaacctgc tgacccacgt gcagggggat
     1141 ccaagcttgc caccatgccc aagaagaaga ggaaggtgtc caatctcctg actgttcacc
     1201 agaacctccc tgcgctgcca gtagatgcca ctagcgatga ggtcaggaaa aatctcatgg
     1261 atatgtttag ggatagacag gcgttttctg aacacacctg gaaaatgctg cttagcgtgt
     1321 gccgatcctg ggcagcctgg tgtaagctga acaatcgcaa atggttcccc gccgagccgg
     1381 aggacgtgcg cgattacctg ctgtatctcc aggcaagagg gctggctgtc aagactatcc
     1441 agcagcactt gggccaactg aatatgctgc atcgacgcag cgggctcccc cggcctagcg
     1501 attcaaacgc agtctccctt gttatgagga gaattagaaa ggaaaacgta gatgcgggtg
     1561 agagggctaa gcaggctctc gcttttgagc ggactgattt cgaccaggtc agatccctga
     1621 tggagaacag cgatcggtgc caggacatca ggaacctcgc atttctggga attgcatata
     1681 acacacttct gcgcatagct gagatcgccc ggatcagagt gaaagacatc agtcgaacgg
     1741 acggcggccg gatgcttatt catattggac gcacaaagac attggtcagc accgctggcg
     1801 ttgaaaaggc cttgtccctg ggcgtaacga agctggtgga aagatggatc tcagtgtccg
     1861 gcgtggctga cgaccctaat aattacttgt tctgtcgagt gagaaaaaac ggagtcgccg
     1921 cgccctctgc caccagccaa ttgagtacac gggcccttga agggatcttt gaggcaaccc
     1981 accgactcat atacggagcc aaggatgaca gtggccagag gtatctcgcc tggtcaggtc
     2041 attctgctag ggtgggggcc gcacgagaca tggcgcgggc aggagtctcc ataccagaga
     2101 ttatgcaagc tggaggttgg acaaatgtga acatcgttat gaactatatc cgcaatcttg
     2161 actctgaaac cggggccatg gtgagactgc tcgaagatgg tgactaccca tacgatgttc
     2221 ctgactatgc gggctatccc tatgacgtcc cggactatgc aggatcctat ccatatgacg
     2281 ttccagatta cgcttgagcg gccgcgaatt cgatatcaag cttatcgata atcaacctct
     2341 ggattacaaa atttgtgaaa gattgactgg tattcttaac tatgttgctc cttttacgct
     2401 atgtggatac gctgctttaa tgcctttgta tcatgctatt gcttcccgta tggctttcat
     2461 tttctcctcc ttgtataaat cctggttgct gtctctttat gaggagttgt ggcccgttgt
     2521 caggcaacgt ggcgtggtgt gcactgtgtt tgctgacgca acccccactg gttggggcat
     2581 tgccaccacc tgtcagctcc tttccgggac tttcgctttc cccctcccta ttgccacggc
     2641 ggaactcatc gccgcctgcc ttgcccgctg ctggacaggg gctcggctgt tgggcactga
     2701 caattccgtg gtgttgtcgg ggaaatcatc gtcctttcct tggctgctcg cctatgttgc
     2761 cacctggatt ctgcgcggga cgtccttctg ctacgtccct tcggccctca atccagcgga
     2821 ccttccttcc cgcggcctgc tgccggctct gcggcctctt ccgcgtcttc gccttcgccc
     2881 tcagacgagt cggatctccc tttgggccgc ctccccgcat cgataccgag cgctgctcga
     2941 gagatctacg ggtggcatcc ctgtgacccc tccccagtgc ctctcctggc cctggaagtt
     3001 gccactccag tgcccaccag ccttgtccta ataaaattaa gttgcatcat tttgtctgac
     3061 taggtgtcct tctataatat tatggggtgg aggggggtgg tatggagcaa ggggcaagtt
     3121 gggaagacaa cctgtagggc ctgcggggtc tattgggaac caagctggag tgcagtggca
     3181 caatcttggc tcactgcaat ctccgcctcc tgggttcaag cgattctcct gcctcagcct
     3241 cccgagttgt tgggattcca ggcatgcatg accaggctca gctaattttt gtttttttgg
     3301 tagagacggg gtttcaccat attggccagg ctggtctcca actcctaatc tcaggtgatc
     3361 tacccacctt ggcctcccaa attgctggga ttacaggcgt gaaccactgc tcccttccct
     3421 gtccttctga ttttgtaggt aaccacgtgc ggaccgagcg gccgcaggaa cccctagtga
     3481 tggagttggc cactccctct ctgcgcgctc gctcgctcac tgaggccggg cgaccaaagg
     3541 tcgcccgacg cccgggcttt gcccgggcgg cctcagtgag cgagcgagcg cgcagctgcc
     3601 tgcaggggcg cctgatgcgg tattttctcc ttacgcatct gtgcggtatt tcacaccgca
     3661 tacgtcaaag caaccatagt acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt
     3721 ggttacgcgc agcgtgaccg ctacacttgc cagcgcctta gcgcccgctc ctttcgcttt
     3781 cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcgggggct
     3841 ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgatttggg
     3901 tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga
     3961 gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca actctatctc
     4021 gggctattct tttgatttat aagggatttt gccgatttcg gtctattggt taaaaaatga
     4081 gctgatttaa caaaaattta acgcgaattt taacaaaata ttaacgttta caattttatg
     4141 gtgcactctc agtacaatct gctctgatgc cgcatagtta agccagcccc gacacccgcc
     4201 aacacccgct gacgcgccct gacgggcttg tctgctcccg gcatccgctt acagacaagc
     4261 tgtgaccgtc tccgggagct gcatgtgtca gaggttttca ccgtcatcac cgaaacgcgc
     4321 gagacgaaag ggcctcgtga tacgcctatt tttataggtt aatgtcatga taataatggt
     4381 ttcttagacg tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt
     4441 tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca
     4501 ataatattga aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc ttattccctt
     4561 ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctggtga aagtaaaaga
     4621 tgctgaagat cagttgggtg cacgagtggg ttacatcgaa ctggatctca acagcggtaa
     4681 gatccttgag agttttcgcc ccgaagaacg ttttccaatg atgagcactt ttaaagttct
     4741 gctatgtggc gcggtattat cccgtattga cgccgggcaa gagcaactcg gtcgccgcat
     4801 acactattct cagaatgact tggttgagta ctcaccagtc acagaaaagc atcttacgga
     4861 tggcatgaca gtaagagaat tatgcagtgc tgccataacc atgagtgata acactgcggc
     4921 caacttactt ctgacaacga tcggaggacc gaaggagcta accgcttttt tgcacaacat
     4981 gggggatcat gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa
     5041 cgacgagcgt gacaccacga tgcctgtagc aatggcaaca acgttgcgca aactattaac
     5101 tggcgaacta cttactctag cttcccggca acaattaata gactggatgg aggcggataa
     5161 agttgcagga ccacttctgc gctcggccct tccggctggc tggtttattg ctgataaatc
     5221 tggagccggt gagcgtgggt ctcgcggtat cattgcagca ctggggccag atggtaagcc
     5281 ctcccgtatc gtagttatct acacgacggg gagtcaggca actatggatg aacgaaatag
     5341 acagatcgct gagataggtg cctcactgat taagcattgg taactgtcag accaagttta
     5401 ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa
     5461 gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc
     5521 gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat
     5581 ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga
     5641 gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt
     5701 tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata
     5761 cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac
     5821 cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg
     5881 ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg
     5941 tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag
     6001 cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct
     6061 ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc
     6121 aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt
     6181 ttgctggcct tttgctcaca t
//
