LOCUS       sequence-469890-        6141 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses the inducible mutant T6B peptide fused with DHFR and YFP 
            under the Purkinje cell-specific L7-6 promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6141)
  AUTHORS   Zolboot N, Xiao Y, Du JX, Ghanem MM, Choi SY, Junn MJ, Zampa F, 
            Huang Z, MacRae IJ, Lippi G
  TITLE     MicroRNA mechanisms instructing Purkinje cell specification.
  JOURNAL   Neuron. 2025 Apr 1:S0896-6273(25)00181-3. doi: 
            10.1016/j.neuron.2025.03.009.
  PUBMED    40179877
REFERENCE   2  (bases 1 to 6141)
  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"; journalName: "Neuron. 
            2025 Apr 1:S0896-6273(25)00181-3. doi: 
            10.1016/j.neuron.2025.03.009."
COMMENT     Sequence Label: pAAV-L7-6-DD-T6B-MUT-YFP
FEATURES             Location/Qualifiers
     source          1..6141
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..141
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   12..141
                     /label=AAV2 ITR
     protein_bind    148..168
                     /gene="mutant version of attB"
                     /label=attB4
                     /bound_moiety="BP Clonase(TM)"
                     /note="core recombination site for the Gateway(R) BP 
                     reaction"
     CDS             join(1805..1807,1808..1810,1811..2524)
                     /codon_start=1
                     /product="enhanced YFP"
                     /label=EYFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1850..1871)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2111..2130)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2458..2479
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     protein_bind    complement(2531..2555)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     misc_feature    2565..3153
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2618..2638)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3036..3047)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(3183..3200)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3189..3396
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   3404..3544
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3404..3533
                     /label=AAV2 ITR
     rep_origin      3619..4074
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3706..3725)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3916..3937
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4091..4110)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4210..4232
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4270..4288)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4356..4460
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4461..4529,4530..5321)
                     /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(4679..4698)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5492..6080
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5981..6000
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcaaag cccgggcgtc
       61 gggcgacctt tggtcgcccg gcctcagtga gcgagcgagc gcgcagagag ggagtggcca
      121 actccatcac taggggttcc ttctagacaa ctttgtatag aaaagttgac tagtggttcc
      181 accctcatgt tggttgatct tcaacattgt cctgacttct tcccacttga cattcctcag
      241 ggtcctgtga tcatggctgg gtctagtgag gttcaaacct gcactgccct acccacaccc
      301 acacccagct cagcgtcagt caggatcaac agttacctag agatcatctt tctggggctt
      361 aagcattggt gggagcagat gggatatgag ttggggattt ggggatgggg gaagatatct
      421 gctccccctc cccctacacc ctagcctttt aaaaggcctt ctcaggtcag agaccaggag
      481 aaaagtatag gagagataca caatggacca ggaagaagaa aagggagagg gaggctcaga
      541 ccttctagac aaggtaagag ggctctggct gactccacca tccgcttctt gaggtctcgg
      601 cacctgtaat tgacaagatt aattcattta tagggcatct aattagcaag taagtctctg
      661 gagtcccctg acccagttac tataacacac agggggtata ggtaggggag tataagagcc
      721 cctcctcagg gcaaatgaat ggattcttag tactgtcccc caagagatag taggtactag
      781 gatttagggg cacttctgag ccccatttcc ctggtaagtg tcccaacccc ccaaatcaac
      841 ccaagcctgg tctcaatcta ggacagtggt agaatgctgt ccctagagtc agtaccatgt
      901 gaaattgtgc tgcaggcagg ggccccaggc tgggaggtgg gggttggggg agtcaggggc
      961 aggtcaggga aggagactca ggtttcattt agagaaattc tgcagacccg tgaggactca
     1021 agtttgtaca aaaaagctcg aggccaccat gatctctctg attgccgctc tggccgtgga
     1081 ctacgtgatc gggatggaaa acgctatgcc atggaatctg cccgccgatc tggcttggtt
     1141 caagaggaac accctgaaca agccagtgat catgggcaga tacacttggg agtccattgg
     1201 ccggcccctg cctggacgca agaacatcat tctgagctcc cagccctcta ccgacgacag
     1261 ggtgacatgg gtgaaaagtg tggacgaagc cattgccgct tgcggagatg tgcccgagat
     1321 catggtcatc ggcggaggga gagtgatcga gcagttcctg cctaaggccc agaaactgta
     1381 cctgactcac attgacgctg aggtggaagg ggacacccat tttcctgatt atgagccaga
     1441 cgattgggaa agcgtgttct ccgagtttca cgacgccgat gctcagaatt ctcatagtta
     1501 ttgctttgag atcctggaaa ggagacgcgg ccgcgattgt caggctgtct tgcagactct
     1561 tttgagccga actgatttgg accccagggt gctctcaaac actggcgccg gccaaactca
     1621 aattaagcag gacacagtgg ccgacattga agaggtgcca aggcctgagg ggaaatctga
     1681 caaaggaact gagggggccg agagcgctgc cacacagacc aagaactcag ggggcgccgg
     1741 agatgcaccc agccaaagca atcaaatgaa gtctggagcc ggggagctcg tcgccaccgg
     1801 atccatggtg agcaagggcg aggagctgtt caccggggtg gtgcccatcc tggtcgagct
     1861 ggacggcgac gtaaacggcc acaagttcag cgtgtccggc gagggcgagg gcgatgccac
     1921 ctacggcaag ctgaccctga agttcatctg caccaccggc aagctgcccg tgccctggcc
     1981 caccctcgtg accaccttcg gctacggcct gcagtgcttc gcccgctacc ccgaccacat
     2041 gaagcagcac gacttcttca agtccgccat gcccgaaggc tacgtccagg agcgcaccat
     2101 cttcttcaag gacgacggca actacaagac ccgcgccgag gtgaagttcg agggcgacac
     2161 cctggtgaac cgcatcgagc tgaagggcat cgacttcaag gaggacggca acatcctggg
     2221 gcacaagctg gagtacaact acaacagcca caacgtctat atcatggccg acaagcagaa
     2281 gaacggcatc aaggtgaact tcaagatccg ccacaacatc gaggacggca gcgtgcagct
     2341 cgccgaccac taccagcaga acacccccat cggcgacggc cccgtgctgc tgcccgacaa
     2401 ccactacctg agctaccagt ccgccctgag caaagacccc aacgagaagc gcgatcacat
     2461 ggtcctgctg gagttcgtga ccgccgccgg gatcactctc ggcatggacg agctgtacaa
     2521 gtaagtcgac acccagcttt cttgtacaaa gtgggaattc cgataatcaa cctctggatt
     2581 acaaaatttg tgaaagattg actggtattc ttaactatgt tgctcctttt acgctatgtg
     2641 gatacgctgc tttaatgcct ttgtatcatg ctattgcttc ccgtatggct ttcattttct
     2701 cctccttgta taaatcctgg ttgctgtctc tttatgagga gttgtggccc gttgtcaggc
     2761 aacgtggcgt ggtgtgcact gtgtttgctg acgcaacccc cactggttgg ggcattgcca
     2821 ccacctgtca gctcctttcc gggactttcg ctttccccct ccctattgcc acggcggaac
     2881 tcatcgccgc ctgccttgcc cgctgctgga caggggctcg gctgttgggc actgacaatt
     2941 ccgtggtgtt gtcggggaag ctgacgtcct ttccatggct gctcgcctgt gttgccacct
     3001 ggattctgcg cgggacgtcc ttctgctacg tcccttcggc cctcaatcca gcggaccttc
     3061 cttcccgcgg cctgctgccg gctctgcggc ctcttccgcg tcttcgcctt cgccctcaga
     3121 cgagtcggat ctccctttgg gccgcctccc cgcatcggga attcctagag ctcgctgatc
     3181 agcctcgact gtgccttcta gttgccagcc atctgttgtt tgcccctccc ccgtgccttc
     3241 cttgaccctg gaaggtgcca ctcccactgt cctttcctaa taaaatgagg aaattgcatc
     3301 gcattgtctg agtaggtgtc attctattct ggggggtggg gtggggcagg acagcaaggg
     3361 ggaggattgg gaagagaata gcaggcatgc tggggagggc cgcaggaacc cctagtgatg
     3421 gagttggcca ctccctctct gcgcgctcgc tcgctcactg aggccgggcg accaaaggtc
     3481 gcccgacgcc cgggctttgc ccgggcggcc tcagtgagcg agcgagcgcg cagctgcctg
     3541 caggggcgcc tgatgcggta ttttctcctt acgcatctgt gcggtatttc acaccgcata
     3601 cgtcaaagca accatagtac gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg
     3661 ttacgcgcag cgtgaccgct acacttgcca gcgccttagc gcccgctcct ttcgctttct
     3721 tcccttcctt tctcgccacg ttcgccggct ttccccgtca agctctaaat cgggggctcc
     3781 ctttagggtt ccgatttagt gctttacggc acctcgaccc caaaaaactt gatttgggtg
     3841 atggttcacg tagtgggcca tcgccctgat agacggtttt tcgccctttg acgttggagt
     3901 ccacgttctt taatagtgga ctcttgttcc aaactggaac aacactcaac tctatctcgg
     3961 gctattcttt tgatttataa gggattttgc cgatttcggt ctattggtta aaaaatgagc
     4021 tgatttaaca aaaatttaac gcgaatttta acaaaatatt aacgtttaca attttatggt
     4081 gcactctcag tacaatctgc tctgatgccg catagttaag ccagccccga cacccgccaa
     4141 cacccgctga cgcgccctga cgggcttgtc tgctcccggc atccgcttac agacaagctg
     4201 tgaccgtctc cgggagctgc atgtgtcaga ggttttcacc gtcatcaccg aaacgcgcga
     4261 gacgaaaggg cctcgtgata cgcctatttt tataggttaa tgtcatgata ataatggttt
     4321 cttagacgtc aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt
     4381 tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat
     4441 aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt
     4501 ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg
     4561 ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga
     4621 tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc
     4681 tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga gcaactcggt cgccgcatac
     4741 actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg
     4801 gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca
     4861 acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg
     4921 gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg
     4981 acgagcgtga caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg
     5041 gcgaactact tactctagct tcccggcaac aattaataga ctggatggag gcggataaag
     5101 ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg
     5161 gagccggtga gcgtggaagc cgcggtatca ttgcagcact ggggccagat ggtaagccct
     5221 cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac
     5281 agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac caagtttact
     5341 catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga
     5401 tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt
     5461 cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct
     5521 gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc
     5581 taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc
     5641 ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc
     5701 tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg
     5761 ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt
     5821 cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg
     5881 agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg
     5941 gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt
     6001 atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag
     6061 gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt
     6121 gctggccttt tgctcacatg t
//
