LOCUS       sequence-469835-        5328 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses the T6B peptide fused with YFP under the human synapsin 
            promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5328)
  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 5328)
  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-FLAG-HA-T6B-WT-YFP
FEATURES             Location/Qualifiers
     source          1..5328
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      1..589
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     490..509
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   651..791
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   662..791
                     /label=AAV2 ITR
     protein_bind    798..818
                     /gene="mutant version of attB"
                     /label=attB4
                     /bound_moiety="BP Clonase(TM)"
                     /note="core recombination site for the Gateway(R) BP 
                     reaction"
     promoter        825..1272
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     regulatory      1303..1312
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1312..1335
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             1336..1362
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             join(1642..1644,1645..1647,1648..2361)
                     /codon_start=1
                     /product="enhanced YFP"
                     /label=EYFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1687..1708)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1948..1967)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2295..2316
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     protein_bind    complement(2368..2392)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     misc_feature    2402..2990
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2455..2475)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2873..2884)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(3020..3037)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3026..3233
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   3241..3381
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3241..3370
                     /label=AAV2 ITR
     rep_origin      3456..3911
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3543..3562)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3753..3774
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3928..3947)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4047..4069
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4107..4125)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4193..4297
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4298..4366,4367..5158)
                     /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(4516..4535)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
       61 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
      121 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
      181 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
      241 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
      301 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
      361 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
      421 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
      481 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
      541 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
      601 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt cctgcaggca
      661 gctgcgcgct cgctcgctca ctgaggccgc ccgggcaaag cccgggcgtc gggcgacctt
      721 tggtcgcccg gcctcagtga gcgagcgagc gcgcagagag ggagtggcca actccatcac
      781 taggggttcc ttctagacaa ctttgtatag aaaagttgac tagtagtgca agtgggtttt
      841 aggaccagga tgaggcgggg tgggggtgcc tacctgacga ccgaccccga cccactggac
      901 aagcacccaa cccccattcc ccaaattgcg catcccctat cagagagggg gaggggaaac
      961 aggatgcggc gaggcgcgtg cgcactgcca gcttcagcac cgcggacagt gccttcgccc
     1021 ccgcctggcg gcgcgcgcca ccgccgcctc agcactgaag gcgcgctgac gtcactcgcc
     1081 ggtcccccgc aaactcccct tcccggccac cttggtcgcg tccgcgccgc cgccggccca
     1141 gccggaccgc accacgcgag gcgcgagata ggggggcacg ggcgcgacca tctgcgctgc
     1201 ggcgccggcg actcagcgct gcctcagtct gcggtgggca gcggaggagt cgtgtcgtgc
     1261 ctgagagcgc agtcgatcta gacgataagg gcgaatctcg aggccaccat ggactacaag
     1321 gacgacgatg acaagtaccc ttatgacgtg cccgattacg ctagcggccg cgattgtcag
     1381 gctgtcttgc agactctttt gagccgaact gatttggacc ccagggtgct ctcaaacact
     1441 ggctggggcc aaactcaaat taagcaggac acagtgtggg acattgaaga ggtgccaagg
     1501 cctgagggga aatctgacaa aggaactgag gggtgggaga gcgctgccac acagaccaag
     1561 aactcagggg gctggggaga tgcacccagc caaagcaatc aaatgaagtc tggatggggg
     1621 gagctcgtcg ccaccggatc catggtgagc aagggcgagg agctgttcac cggggtggtg
     1681 cccatcctgg tcgagctgga cggcgacgta aacggccaca agttcagcgt gtccggcgag
     1741 ggcgagggcg atgccaccta cggcaagctg accctgaagt tcatctgcac caccggcaag
     1801 ctgcccgtgc cctggcccac cctcgtgacc accttcggct acggcctgca gtgcttcgcc
     1861 cgctaccccg accacatgaa gcagcacgac ttcttcaagt ccgccatgcc cgaaggctac
     1921 gtccaggagc gcaccatctt cttcaaggac gacggcaact acaagacccg cgccgaggtg
     1981 aagttcgagg gcgacaccct ggtgaaccgc atcgagctga agggcatcga cttcaaggag
     2041 gacggcaaca tcctggggca caagctggag tacaactaca acagccacaa cgtctatatc
     2101 atggccgaca agcagaagaa cggcatcaag gtgaacttca agatccgcca caacatcgag
     2161 gacggcagcg tgcagctcgc cgaccactac cagcagaaca cccccatcgg cgacggcccc
     2221 gtgctgctgc ccgacaacca ctacctgagc taccagtccg ccctgagcaa agaccccaac
     2281 gagaagcgcg atcacatggt cctgctggag ttcgtgaccg ccgccgggat cactctcggc
     2341 atggacgagc tgtacaagta agtcgacacc cagctttctt gtacaaagtg ggaattccga
     2401 taatcaacct ctggattaca aaatttgtga aagattgact ggtattctta actatgttgc
     2461 tccttttacg ctatgtggat acgctgcttt aatgcctttg tatcatgcta ttgcttcccg
     2521 tatggctttc attttctcct ccttgtataa atcctggttg ctgtctcttt atgaggagtt
     2581 gtggcccgtt gtcaggcaac gtggcgtggt gtgcactgtg tttgctgacg caacccccac
     2641 tggttggggc attgccacca cctgtcagct cctttccggg actttcgctt tccccctccc
     2701 tattgccacg gcggaactca tcgccgcctg ccttgcccgc tgctggacag gggctcggct
     2761 gttgggcact gacaattccg tggtgttgtc ggggaagctg acgtcctttc catggctgct
     2821 cgcctgtgtt gccacctgga ttctgcgcgg gacgtccttc tgctacgtcc cttcggccct
     2881 caatccagcg gaccttcctt cccgcggcct gctgccggct ctgcggcctc ttccgcgtct
     2941 tcgccttcgc cctcagacga gtcggatctc cctttgggcc gcctccccgc atcgggaatt
     3001 cctagagctc gctgatcagc ctcgactgtg ccttctagtt gccagccatc tgttgtttgc
     3061 ccctcccccg tgccttcctt gaccctggaa ggtgccactc ccactgtcct ttcctaataa
     3121 aatgaggaaa ttgcatcgca ttgtctgagt aggtgtcatt ctattctggg gggtggggtg
     3181 gggcaggaca gcaaggggga ggattgggaa gagaatagca ggcatgctgg ggagggccgc
     3241 aggaacccct agtgatggag ttggccactc cctctctgcg cgctcgctcg ctcactgagg
     3301 ccgggcgacc aaaggtcgcc cgacgcccgg gctttgcccg ggcggcctca gtgagcgagc
     3361 gagcgcgcag ctgcctgcag gggcgcctga tgcggtattt tctccttacg catctgtgcg
     3421 gtatttcaca ccgcatacgt caaagcaacc atagtacgcg ccctgtagcg gcgcattaag
     3481 cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccttagcgcc
     3541 cgctcctttc gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc
     3601 tctaaatcgg gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa
     3661 aaaacttgat ttgggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg
     3721 ccctttgacg ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac
     3781 actcaactct atctcgggct attcttttga tttataaggg attttgccga tttcggtcta
     3841 ttggttaaaa aatgagctga tttaacaaaa atttaacgcg aattttaaca aaatattaac
     3901 gtttacaatt ttatggtgca ctctcagtac aatctgctct gatgccgcat agttaagcca
     3961 gccccgacac ccgccaacac ccgctgacgc gccctgacgg gcttgtctgc tcccggcatc
     4021 cgcttacaga caagctgtga ccgtctccgg gagctgcatg tgtcagaggt tttcaccgtc
     4081 atcaccgaaa cgcgcgagac gaaagggcct cgtgatacgc ctatttttat aggttaatgt
     4141 catgataata atggtttctt agacgtcagg tggcactttt cggggaaatg tgcgcggaac
     4201 ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc
     4261 ctgataaatg cttcaataat attgaaaaag gaagagtatg agtattcaac atttccgtgt
     4321 cgcccttatt cccttttttg cggcattttg ccttcctgtt tttgctcacc cagaaacgct
     4381 ggtgaaagta aaagatgctg aagatcagtt gggtgcacga gtgggttaca tcgaactgga
     4441 tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc caatgatgag
     4501 cacttttaaa gttctgctat gtggcgcggt attatcccgt attgacgccg ggcaagagca
     4561 actcggtcgc cgcatacact attctcagaa tgacttggtt gagtactcac cagtcacaga
     4621 aaagcatctt acggatggca tgacagtaag agaattatgc agtgctgcca taaccatgag
     4681 tgataacact gcggccaact tacttctgac aacgatcgga ggaccgaagg agctaaccgc
     4741 ttttttgcac aacatggggg atcatgtaac tcgccttgat cgttgggaac cggagctgaa
     4801 tgaagccata ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg caacaacgtt
     4861 gcgcaaacta ttaactggcg aactacttac tctagcttcc cggcaacaat taatagactg
     4921 gatggaggcg gataaagttg caggaccact tctgcgctcg gcccttccgg ctggctggtt
     4981 tattgctgat aaatctggag ccggtgagcg tggaagccgc ggtatcattg cagcactggg
     5041 gccagatggt aagccctccc gtatcgtagt tatctacacg acggggagtc aggcaactat
     5101 ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc attggtaact
     5161 gtcagaccaa gtttactcat atatacttta gattgattta aaacttcatt tttaatttaa
     5221 aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt
     5281 ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttc
//
