LOCUS       sequence-469892-        5751 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses the inducible T6B peptide fused with DHFR and YFP under 
            the human synapsin promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5751)
  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 5751)
  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-hSyn-DD-T6B-WT-YFP
FEATURES             Location/Qualifiers
     source          1..5751
                     /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"
     promoter        175..622
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     CDS             join(1415..1417,1418..1420,1421..2134)
                     /codon_start=1
                     /product="enhanced YFP"
                     /label=EYFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1460..1481)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1721..1740)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2068..2089
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     protein_bind    complement(2141..2165)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     misc_feature    2175..2763
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2228..2248)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2646..2657)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(2793..2810)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2799..3006
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   3014..3154
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3014..3143
                     /label=AAV2 ITR
     rep_origin      3229..3684
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3316..3335)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3526..3547
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3701..3720)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3820..3842
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3880..3898)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3966..4070
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4071..4139,4140..4931)
                     /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(4289..4308)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5102..5690
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5591..5610
                     /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 tagtagtgca
      181 agtgggtttt aggaccagga tgaggcgggg tgggggtgcc tacctgacga ccgaccccga
      241 cccactggac aagcacccaa cccccattcc ccaaattgcg catcccctat cagagagggg
      301 gaggggaaac aggatgcggc gaggcgcgtg cgcactgcca gcttcagcac cgcggacagt
      361 gccttcgccc ccgcctggcg gcgcgcgcca ccgccgcctc agcactgaag gcgcgctgac
      421 gtcactcgcc ggtcccccgc aaactcccct tcccggccac cttggtcgcg tccgcgccgc
      481 cgccggccca gccggaccgc accacgcgag gcgcgagata ggggggcacg ggcgcgacca
      541 tctgcgctgc ggcgccggcg actcagcgct gcctcagtct gcggtgggca gcggaggagt
      601 cgtgtcgtgc ctgagagcgc agtcgatcta gacgataagg gcgaatctcg aggccaccat
      661 gatctctctg attgccgctc tggccgtgga ctacgtgatc gggatggaaa acgctatgcc
      721 atggaatctg cccgccgatc tggcttggtt caagaggaac accctgaaca agccagtgat
      781 catgggcaga tacacttggg agtccattgg ccggcccctg cctggacgca agaacatcat
      841 tctgagctcc cagccctcta ccgacgacag ggtgacatgg gtgaaaagtg tggacgaagc
      901 cattgccgct tgcggagatg tgcccgagat catggtcatc ggcggaggga gagtgatcga
      961 gcagttcctg cctaaggccc agaaactgta cctgactcac attgacgctg aggtggaagg
     1021 ggacacccat tttcctgatt atgagccaga cgattgggaa agcgtgttct ccgagtttca
     1081 cgacgccgat gctcagaatt ctcatagtta ttgctttgag atcctggaaa ggagacgcgg
     1141 ccgcgattgt caggctgtct tgcagactct tttgagccga actgatttgg accccagggt
     1201 gctctcaaac actggctggg gccaaactca aattaagcag gacacagtgt gggacattga
     1261 agaggtgcca aggcctgagg ggaaatctga caaaggaact gaggggtggg agagcgctgc
     1321 cacacagacc aagaactcag ggggctgggg agatgcaccc agccaaagca atcaaatgaa
     1381 gtctggatgg ggggagctcg tcgccaccgg atccatggtg agcaagggcg aggagctgtt
     1441 caccggggtg gtgcccatcc tggtcgagct ggacggcgac gtaaacggcc acaagttcag
     1501 cgtgtccggc gagggcgagg gcgatgccac ctacggcaag ctgaccctga agttcatctg
     1561 caccaccggc aagctgcccg tgccctggcc caccctcgtg accaccttcg gctacggcct
     1621 gcagtgcttc gcccgctacc ccgaccacat gaagcagcac gacttcttca agtccgccat
     1681 gcccgaaggc tacgtccagg agcgcaccat cttcttcaag gacgacggca actacaagac
     1741 ccgcgccgag gtgaagttcg agggcgacac cctggtgaac cgcatcgagc tgaagggcat
     1801 cgacttcaag gaggacggca acatcctggg gcacaagctg gagtacaact acaacagcca
     1861 caacgtctat atcatggccg acaagcagaa gaacggcatc aaggtgaact tcaagatccg
     1921 ccacaacatc gaggacggca gcgtgcagct cgccgaccac taccagcaga acacccccat
     1981 cggcgacggc cccgtgctgc tgcccgacaa ccactacctg agctaccagt ccgccctgag
     2041 caaagacccc aacgagaagc gcgatcacat ggtcctgctg gagttcgtga ccgccgccgg
     2101 gatcactctc ggcatggacg agctgtacaa gtaagtcgac acccagcttt cttgtacaaa
     2161 gtgggaattc cgataatcaa cctctggatt acaaaatttg tgaaagattg actggtattc
     2221 ttaactatgt tgctcctttt acgctatgtg gatacgctgc tttaatgcct ttgtatcatg
     2281 ctattgcttc ccgtatggct ttcattttct cctccttgta taaatcctgg ttgctgtctc
     2341 tttatgagga gttgtggccc gttgtcaggc aacgtggcgt ggtgtgcact gtgtttgctg
     2401 acgcaacccc cactggttgg ggcattgcca ccacctgtca gctcctttcc gggactttcg
     2461 ctttccccct ccctattgcc acggcggaac tcatcgccgc ctgccttgcc cgctgctgga
     2521 caggggctcg gctgttgggc actgacaatt ccgtggtgtt gtcggggaag ctgacgtcct
     2581 ttccatggct gctcgcctgt gttgccacct ggattctgcg cgggacgtcc ttctgctacg
     2641 tcccttcggc cctcaatcca gcggaccttc cttcccgcgg cctgctgccg gctctgcggc
     2701 ctcttccgcg tcttcgcctt cgccctcaga cgagtcggat ctccctttgg gccgcctccc
     2761 cgcatcggga attcctagag ctcgctgatc agcctcgact gtgccttcta gttgccagcc
     2821 atctgttgtt tgcccctccc ccgtgccttc cttgaccctg gaaggtgcca ctcccactgt
     2881 cctttcctaa taaaatgagg aaattgcatc gcattgtctg agtaggtgtc attctattct
     2941 ggggggtggg gtggggcagg acagcaaggg ggaggattgg gaagagaata gcaggcatgc
     3001 tggggagggc cgcaggaacc cctagtgatg gagttggcca ctccctctct gcgcgctcgc
     3061 tcgctcactg aggccgggcg accaaaggtc gcccgacgcc cgggctttgc ccgggcggcc
     3121 tcagtgagcg agcgagcgcg cagctgcctg caggggcgcc tgatgcggta ttttctcctt
     3181 acgcatctgt gcggtatttc acaccgcata cgtcaaagca accatagtac gcgccctgta
     3241 gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca
     3301 gcgccttagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg ttcgccggct
     3361 ttccccgtca agctctaaat cgggggctcc ctttagggtt ccgatttagt gctttacggc
     3421 acctcgaccc caaaaaactt gatttgggtg atggttcacg tagtgggcca tcgccctgat
     3481 agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga ctcttgttcc
     3541 aaactggaac aacactcaac tctatctcgg gctattcttt tgatttataa gggattttgc
     3601 cgatttcggt ctattggtta aaaaatgagc tgatttaaca aaaatttaac gcgaatttta
     3661 acaaaatatt aacgtttaca attttatggt gcactctcag tacaatctgc tctgatgccg
     3721 catagttaag ccagccccga cacccgccaa cacccgctga cgcgccctga cgggcttgtc
     3781 tgctcccggc atccgcttac agacaagctg tgaccgtctc cgggagctgc atgtgtcaga
     3841 ggttttcacc gtcatcaccg aaacgcgcga gacgaaaggg cctcgtgata cgcctatttt
     3901 tataggttaa tgtcatgata ataatggttt cttagacgtc aggtggcact tttcggggaa
     3961 atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca
     4021 tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt atgagtattc
     4081 aacatttccg tgtcgccctt attccctttt ttgcggcatt ttgccttcct gtttttgctc
     4141 acccagaaac gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt
     4201 acatcgaact ggatctcaac agcggtaaga tccttgagag ttttcgcccc gaagaacgtt
     4261 ttccaatgat gagcactttt aaagttctgc tatgtggcgc ggtattatcc cgtattgacg
     4321 ccgggcaaga gcaactcggt cgccgcatac actattctca gaatgacttg gttgagtact
     4381 caccagtcac agaaaagcat cttacggatg gcatgacagt aagagaatta tgcagtgctg
     4441 ccataaccat gagtgataac actgcggcca acttacttct gacaacgatc ggaggaccga
     4501 aggagctaac cgcttttttg cacaacatgg gggatcatgt aactcgcctt gatcgttggg
     4561 aaccggagct gaatgaagcc ataccaaacg acgagcgtga caccacgatg cctgtagcaa
     4621 tggcaacaac gttgcgcaaa ctattaactg gcgaactact tactctagct tcccggcaac
     4681 aattaataga ctggatggag gcggataaag ttgcaggacc acttctgcgc tcggcccttc
     4741 cggctggctg gtttattgct gataaatctg gagccggtga gcgtggaagc cgcggtatca
     4801 ttgcagcact ggggccagat ggtaagccct cccgtatcgt agttatctac acgacgggga
     4861 gtcaggcaac tatggatgaa cgaaatagac agatcgctga gataggtgcc tcactgatta
     4921 agcattggta actgtcagac caagtttact catatatact ttagattgat ttaaaacttc
     4981 atttttaatt taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc
     5041 cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt
     5101 cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac
     5161 cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct
     5221 tcagcagagc gcagatacca aatactgttc ttctagtgta gccgtagtta ggccaccact
     5281 tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg
     5341 ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata
     5401 aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga
     5461 cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag
     5521 ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg
     5581 agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac
     5641 ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca
     5701 acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg t
//
