LOCUS       sequence-469893-        5729 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses the inducible mutant 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 5729)
  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 5729)
  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-MUT-YFP
FEATURES             Location/Qualifiers
     source          1..5729
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    126..146
                     /gene="mutant version of attB"
                     /label=attB4
                     /bound_moiety="BP Clonase(TM)"
                     /note="core recombination site for the Gateway(R) BP 
                     reaction"
     promoter        153..600
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     CDS             join(1393..1395,1396..1398,1399..2112)
                     /codon_start=1
                     /product="enhanced YFP"
                     /label=EYFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1438..1459)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1699..1718)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2046..2067
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     protein_bind    complement(2119..2143)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     misc_feature    2153..2741
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2206..2226)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2624..2635)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(2771..2788)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2777..2984
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   2992..3132
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2992..3121
                     /label=AAV2 ITR
     rep_origin      3207..3662
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3294..3313)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3504..3525
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3679..3698)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3798..3820
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3858..3876)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3944..4048
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4049..4117,4118..4909)
                     /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(4267..4286)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5080..5668
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5569..5588
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcaaag cccgggcggc
       61 ctcagtgagc gagcgagcgc gcagagaggg agtggccaac tccatcacta ggggttcctt
      121 ctagacaact ttgtatagaa aagttgacta gtagtgcaag tgggttttag gaccaggatg
      181 aggcggggtg ggggtgccta cctgacgacc gaccccgacc cactggacaa gcacccaacc
      241 cccattcccc aaattgcgca tcccctatca gagaggggga ggggaaacag gatgcggcga
      301 ggcgcgtgcg cactgccagc ttcagcaccg cggacagtgc cttcgccccc gcctggcggc
      361 gcgcgccacc gccgcctcag cactgaaggc gcgctgacgt cactcgccgg tcccccgcaa
      421 actccccttc ccggccacct tggtcgcgtc cgcgccgccg ccggcccagc cggaccgcac
      481 cacgcgaggc gcgagatagg ggggcacggg cgcgaccatc tgcgctgcgg cgccggcgac
      541 tcagcgctgc ctcagtctgc ggtgggcagc ggaggagtcg tgtcgtgcct gagagcgcag
      601 tcgatctaga cgataagggc gaatctcgag gccaccatga tctctctgat tgccgctctg
      661 gccgtggact acgtgatcgg gatggaaaac gctatgccat ggaatctgcc cgccgatctg
      721 gcttggttca agaggaacac cctgaacaag ccagtgatca tgggcagata cacttgggag
      781 tccattggcc ggcccctgcc tggacgcaag aacatcattc tgagctccca gccctctacc
      841 gacgacaggg tgacatgggt gaaaagtgtg gacgaagcca ttgccgcttg cggagatgtg
      901 cccgagatca tggtcatcgg cggagggaga gtgatcgagc agttcctgcc taaggcccag
      961 aaactgtacc tgactcacat tgacgctgag gtggaagggg acacccattt tcctgattat
     1021 gagccagacg attgggaaag cgtgttctcc gagtttcacg acgccgatgc tcagaattct
     1081 catagttatt gctttgagat cctggaaagg agacgcggcc gcgattgtca ggctgtcttg
     1141 cagactcttt tgagccgaac tgatttggac cccagggtgc tctcaaacac tggcgccggc
     1201 caaactcaaa ttaagcagga cacagtggcc gacattgaag aggtgccaag gcctgagggg
     1261 aaatctgaca aaggaactga gggggccgag agcgctgcca cacagaccaa gaactcaggg
     1321 ggcgccggag atgcacccag ccaaagcaat caaatgaagt ctggagccgg ggagctcgtc
     1381 gccaccggat ccatggtgag caagggcgag gagctgttca ccggggtggt gcccatcctg
     1441 gtcgagctgg acggcgacgt aaacggccac aagttcagcg tgtccggcga gggcgagggc
     1501 gatgccacct acggcaagct gaccctgaag ttcatctgca ccaccggcaa gctgcccgtg
     1561 ccctggccca ccctcgtgac caccttcggc tacggcctgc agtgcttcgc ccgctacccc
     1621 gaccacatga agcagcacga cttcttcaag tccgccatgc ccgaaggcta cgtccaggag
     1681 cgcaccatct tcttcaagga cgacggcaac tacaagaccc gcgccgaggt gaagttcgag
     1741 ggcgacaccc tggtgaaccg catcgagctg aagggcatcg acttcaagga ggacggcaac
     1801 atcctggggc acaagctgga gtacaactac aacagccaca acgtctatat catggccgac
     1861 aagcagaaga acggcatcaa ggtgaacttc aagatccgcc acaacatcga ggacggcagc
     1921 gtgcagctcg ccgaccacta ccagcagaac acccccatcg gcgacggccc cgtgctgctg
     1981 cccgacaacc actacctgag ctaccagtcc gccctgagca aagaccccaa cgagaagcgc
     2041 gatcacatgg tcctgctgga gttcgtgacc gccgccggga tcactctcgg catggacgag
     2101 ctgtacaagt aagtcgacac ccagctttct tgtacaaagt gggaattccg ataatcaacc
     2161 tctggattac aaaatttgtg aaagattgac tggtattctt aactatgttg ctccttttac
     2221 gctatgtgga tacgctgctt taatgccttt gtatcatgct attgcttccc gtatggcttt
     2281 cattttctcc tccttgtata aatcctggtt gctgtctctt tatgaggagt tgtggcccgt
     2341 tgtcaggcaa cgtggcgtgg tgtgcactgt gtttgctgac gcaaccccca ctggttgggg
     2401 cattgccacc acctgtcagc tcctttccgg gactttcgct ttccccctcc ctattgccac
     2461 ggcggaactc atcgccgcct gccttgcccg ctgctggaca ggggctcggc tgttgggcac
     2521 tgacaattcc gtggtgttgt cggggaagct gacgtccttt ccatggctgc tcgcctgtgt
     2581 tgccacctgg attctgcgcg ggacgtcctt ctgctacgtc ccttcggccc tcaatccagc
     2641 ggaccttcct tcccgcggcc tgctgccggc tctgcggcct cttccgcgtc ttcgccttcg
     2701 ccctcagacg agtcggatct ccctttgggc cgcctccccg catcgggaat tcctagagct
     2761 cgctgatcag cctcgactgt gccttctagt tgccagccat ctgttgtttg cccctccccc
     2821 gtgccttcct tgaccctgga aggtgccact cccactgtcc tttcctaata aaatgaggaa
     2881 attgcatcgc attgtctgag taggtgtcat tctattctgg ggggtggggt ggggcaggac
     2941 agcaaggggg aggattggga agagaatagc aggcatgctg gggagggccg caggaacccc
     3001 tagtgatgga gttggccact ccctctctgc gcgctcgctc gctcactgag gccgggcgac
     3061 caaaggtcgc ccgacgcccg ggctttgccc gggcggcctc agtgagcgag cgagcgcgca
     3121 gctgcctgca ggggcgcctg atgcggtatt ttctccttac gcatctgtgc ggtatttcac
     3181 accgcatacg tcaaagcaac catagtacgc gccctgtagc ggcgcattaa gcgcggcggg
     3241 tgtggtggtt acgcgcagcg tgaccgctac acttgccagc gccttagcgc ccgctccttt
     3301 cgctttcttc ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg
     3361 ggggctccct ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga
     3421 tttgggtgat ggttcacgta gtgggccatc gccctgatag acggtttttc gccctttgac
     3481 gttggagtcc acgttcttta atagtggact cttgttccaa actggaacaa cactcaactc
     3541 tatctcgggc tattcttttg atttataagg gattttgccg atttcggtct attggttaaa
     3601 aaatgagctg atttaacaaa aatttaacgc gaattttaac aaaatattaa cgtttacaat
     3661 tttatggtgc actctcagta caatctgctc tgatgccgca tagttaagcc agccccgaca
     3721 cccgccaaca cccgctgacg cgccctgacg ggcttgtctg ctcccggcat ccgcttacag
     3781 acaagctgtg accgtctccg ggagctgcat gtgtcagagg ttttcaccgt catcaccgaa
     3841 acgcgcgaga cgaaagggcc tcgtgatacg cctattttta taggttaatg tcatgataat
     3901 aatggtttct tagacgtcag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg
     3961 tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat
     4021 gcttcaataa tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat
     4081 tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt
     4141 aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag
     4201 cggtaagatc cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa
     4261 agttctgcta tgtggcgcgg tattatcccg tattgacgcc gggcaagagc aactcggtcg
     4321 ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct
     4381 tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac
     4441 tgcggccaac ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca
     4501 caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat
     4561 accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact
     4621 attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc
     4681 ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga
     4741 taaatctgga gccggtgagc gtggaagccg cggtatcatt gcagcactgg ggccagatgg
     4801 taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg
     4861 aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca
     4921 agtttactca tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta
     4981 ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca
     5041 ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg
     5101 cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga
     5161 tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa
     5221 tactgttctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc
     5281 tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg
     5341 tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac
     5401 ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct
     5461 acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc
     5521 ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg
     5581 gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg
     5641 ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct
     5701 ggccttttgc tggccttttg ctcacatgt
//
