LOCUS       sequence-237547-        6295 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Allows for initial cellular transduction of EnvA-pseudotyped Rb via 
            Cre-dependent expression of an avian TVA receptor fused with yellow 
            fluorescent protein.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6295)
  AUTHORS   Faget L, Osakada F, Duan J, Ressler R, Johnson AB, Proudfoot JA, Yoo
            JH, Callaway EM, Hnasko TS
  TITLE     Afferent Inputs to Neurotransmitter-Defined Cell Types in the 
            Ventral Tegmental Area.
  JOURNAL   Cell Rep. 2016 Jun 21;15(12):2796-808. doi: 
            10.1016/j.celrep.2016.05.057. Epub 2016 Jun 9.
  PUBMED    27292633
REFERENCE   2  (bases 1 to 6295)
  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: 
            "10.1016/j.celrep.2016.05.057"; journalName: "Cell Rep"; date: 
            "2016-06-21- 21"; volume: "15"; issue: "12"; pages: "2796-808"
COMMENT     Sequence Label: pAAV-EnhancedSynapsin-DIO-TVA950-EYFP-WPRE
FEATURES             Location/Qualifiers
     source          1..6295
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   51..180
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     enhancer        255..558
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     protein_bind    complement(991..1024)
                     /label=lox2272
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (AAGTATCC) (Shaw et al., 2021). lox2272 sites are 
                     compatible with each other, but incompatible with loxP or 
                     loxN sites (Lee and Saito, 1988)."
     protein_bind    complement(1075..1108)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     CDS             complement(join(1117..1830,1831..1833,1834..1836))
                     /codon_start=1
                     /product="enhanced YFP"
                     /label=EYFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1162..1183)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     1511..1530
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1770..1791
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      1833..1842
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     regulatory      2316..2325
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    2330..2363
                     /label=lox2272
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (AAGTATCC) (Shaw et al., 2021). lox2272 sites are 
                     compatible with each other, but incompatible with loxP or 
                     loxN sites (Lee and Saito, 1988)."
     protein_bind    2414..2447
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     misc_feature    2472..3060
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2525..2545)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2943..2954)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3092..3568
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(3291..3310)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   3608..3748
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3608..3737
                     /label=AAV2 ITR
     rep_origin      3823..4278
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3910..3929)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4120..4141
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4295..4314)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4414..4436
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4474..4492)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4560..4664
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4665..4733,4734..5525)
                     /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(4883..4902)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5696..6284
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6185..6204
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt cctgcaggca
       61 gctgcgcgct cgctcgctca ctgaggccgc ccgggcgtcg ggcgaccttt ggtcgcccgg
      121 cctcagtgag cgagcgagcg cgcagagagg gagtggccaa ctccatcact aggggttcct
      181 gcggccgcac gcgttagtta ttaatagtaa tcaattacgg ggtcattagt tcatagccca
      241 tatatggagt tccgcgttac ataacttacg gtaaatggcc cgcctggctg accgcccaac
      301 gacccccgcc cattgacgtc aataatgacg tatgttccca tagtaacgcc aatagggact
      361 ttccattgac gtcaatgggt ggagtattta cggtaaactg cccacttggc agtacatcaa
      421 gtgtatcata tgccaagtac gccccctatt gacgtcaatg acggtaaatg gcccgcctgg
      481 cattatgccc agtacatgac cttatgggac tttcctactt ggcagtacat ctacgtatta
      541 gtcatcgcta ttaccatgga gatctctgca gagggccctg cgtatgagtg caagtgggtt
      601 ttaggaccag gatgaggcgg ggtgggggtg cctacctgac gaccgacccc gacccactgg
      661 acaagcaccc aacccccatt ccccaaattg cgcatcccct atcagagagg gggaggggaa
      721 acaggatgcg gcgaggcgcg tgcgcactgc cagcttcagc accgcggaca gtgccttcgc
      781 ccccgcctgg cggcgcgcgc caccgccgcc tcagcactga aggcgcgctg acgtcactcg
      841 ccggtccccc gcaaactccc cttcccggcc accttggtcg cgtccgcgcc gccgccggcc
      901 cagccggacc gcaccacgcg aggcgcgaga taggggggca cgggcgcgac catctgcgct
      961 gcggcgggat cctctagagt cgactccgga ataacttcgt ataggatact ttatacgaag
     1021 ttatgcagaa tggtagctgg attgtagctg ctattagcaa tatgaaacct cttaataact
     1081 tcgtatagca tacattatac gaagttatgg cgcgccttac ttgtacagct cgtccatgcc
     1141 gagagtgatc ccggcggcgg tcacgaactc cagcaggacc atgtgatcgc gcttctcgtt
     1201 ggggtctttg ctcagggcgg actggtagct caggtagtgg ttgtcgggca gcagcacggg
     1261 gccgtcgccg atgggggtgt tctgctggta gtggtcggcg agctgcacgc tgccgtcctc
     1321 gatgttgtgg cggatcttga agttcacctt gatgccgttc ttctgcttgt cggccatgat
     1381 atagacgttg tggctgttgt agttgtactc cagcttgtgc cccaggatgt tgccgtcctc
     1441 cttgaagtcg atgcccttca gctcgatgcg gttcaccagg gtgtcgccct cgaacttcac
     1501 ctcggcgcgg gtcttgtagt tgccgtcgtc cttgaagaag atggtgcgct cctggacgta
     1561 gccttcgggc atggcggact tgaagaagtc gtgctgcttc atgtggtcgg ggtagcgggc
     1621 gaagcactgc aggccgtagc cgaaggtggt cacgagggtg ggccagggca cgggcagctt
     1681 gccggtggtg cagatgaact tcagggtcag cttgccgtag gtggcatcgc cctcgccctc
     1741 gccggacacg ctgaacttgt ggccgtttac gtcgccgtcc agctcgacca ggatgggcac
     1801 caccccggtg aacagctcct cgcccttgct caccatggtg gcggccgcgg agaacaagtc
     1861 tgcctggctc ttgtcaggca ccagcagctc cttggatgcg ctttcaagac tgaagatgtc
     1921 agacctgctt tttgctttgg acttccccca tgcagcgata ccacccacag ctaccaggca
     1981 gcacaggagc actgcagtga tcagcatcca catgcggccg tgattcctgg ctggcagagc
     2041 acgtccagga gcaggggcag tgggagcctc tgtgccgttg tccgtgggca ccgcggggat
     2101 cgcgctggtc ccgcagcccc actcgtcccg cccgtcgtcg cagtcggggt gtccgtcgca
     2161 cagccagtcc tgcgggtaac actccccgtg ggcaccgggc ggctccgagc agcggaactg
     2221 accggggggg caacgggaca aagaaccgtt accggacccg ttaccggtca cgttaccggg
     2281 cagcagcagc agcagcagcg cgggcagcag ccgcgccatg gtggctagca taacttcgta
     2341 taaagtatcc tatacgaagt tatttgcctt aacccagaaa ttatcactgt tattctttag
     2401 aatggtgcaa agaataactt cgtataatgt atgctatacg aagttatgaa ttcgatatca
     2461 agcttatcga taatcaacct ctggattaca aaatttgtga aagattgact ggtattctta
     2521 actatgttgc tccttttacg ctatgtggat acgctgcttt aatgcctttg tatcatgcta
     2581 ttgcttcccg tatggctttc attttctcct ccttgtataa atcctggttg ctgtctcttt
     2641 atgaggagtt gtggcccgtt gtcaggcaac gtggcgtggt gtgcactgtg tttgctgacg
     2701 caacccccac tggttggggc attgccacca cctgtcagct cctttccggg actttcgctt
     2761 tccccctccc tattgccacg gcggaactca tcgccgcctg ccttgcccgc tgctggacag
     2821 gggctcggct gttgggcact gacaattccg tggtgttgtc ggggaaatca tcgtcctttc
     2881 cttggctgct cgcctatgtt gccacctgga ttctgcgcgg gacgtccttc tgctacgtcc
     2941 cttcggccct caatccagcg gaccttcctt cccgcggcct gctgccggct ctgcggcctc
     3001 ttccgcgtct tcgccttcgc cctcagacga gtcggatctc cctttgggcc gcctccccgc
     3061 atcgataccg agcgctgctc gagagatcta cgggtggcat ccctgtgacc cctccccagt
     3121 gcctctcctg gccctggaag ttgccactcc agtgcccacc agccttgtcc taataaaatt
     3181 aagttgcatc attttgtctg actaggtgtc cttctataat attatggggt ggaggggggt
     3241 ggtatggagc aaggggcaag ttgggaagac aacctgtagg gcctgcgggg tctattggga
     3301 accaagctgg agtgcagtgg cacaatcttg gctcactgca atctccgcct cctgggttca
     3361 agcgattctc ctgcctcagc ctcccgagtt gttgggattc caggcatgca tgaccaggct
     3421 cagctaattt ttgttttttt ggtagagacg gggtttcacc atattggcca ggctggtctc
     3481 caactcctaa tctcaggtga tctacccacc ttggcctccc aaattgctgg gattacaggc
     3541 gtgaaccact gctcccttcc ctgtccttct gattttgtag gtaaccacgt gcggaccgag
     3601 cggccgcagg aacccctagt gatggagttg gccactccct ctctgcgcgc tcgctcgctc
     3661 actgaggccg ggcgaccaaa ggtcgcccga cgcccgggct ttgcccgggc ggcctcagtg
     3721 agcgagcgag cgcgcagctg cctgcagggg cgcctgatgc ggtattttct ccttacgcat
     3781 ctgtgcggta tttcacaccg catacgtcaa agcaaccata gtacgcgccc tgtagcggcg
     3841 cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt gccagcgcct
     3901 tagcgcccgc tcctttcgct ttcttccctt cctttctcgc cacgttcgcc ggctttcccc
     3961 gtcaagctct aaatcggggg ctccctttag ggttccgatt tagtgcttta cggcacctcg
     4021 accccaaaaa acttgatttg ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg
     4081 tttttcgccc tttgacgttg gagtccacgt tctttaatag tggactcttg ttccaaactg
     4141 gaacaacact caactctatc tcgggctatt cttttgattt ataagggatt ttgccgattt
     4201 cggtctattg gttaaaaaat gagctgattt aacaaaaatt taacgcgaat tttaacaaaa
     4261 tattaacgtt tacaatttta tggtgcactc tcagtacaat ctgctctgat gccgcatagt
     4321 taagccagcc ccgacacccg ccaacacccg ctgacgcgcc ctgacgggct tgtctgctcc
     4381 cggcatccgc ttacagacaa gctgtgaccg tctccgggag ctgcatgtgt cagaggtttt
     4441 caccgtcatc accgaaacgc gcgagacgaa agggcctcgt gatacgccta tttttatagg
     4501 ttaatgtcat gataataatg gtttcttaga cgtcaggtgg cacttttcgg ggaaatgtgc
     4561 gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac
     4621 aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtatgagt attcaacatt
     4681 tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt gctcacccag
     4741 aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg ggttacatcg
     4801 aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa cgttttccaa
     4861 tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc
     4921 aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag tactcaccag
     4981 tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt gctgccataa
     5041 ccatgagtga taacactgcg gccaacttac ttctgacaac gatcggagga ccgaaggagc
     5101 taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt tgggaaccgg
     5161 agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgta gcaatggcaa
     5221 caacgttgcg caaactatta actggcgaac tacttactct agcttcccgg caacaattaa
     5281 tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc cttccggctg
     5341 gctggtttat tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt atcattgcag
     5401 cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg gggagtcagg
     5461 caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg attaagcatt
     5521 ggtaactgtc agaccaagtt tactcatata tactttagat tgatttaaaa cttcattttt
     5581 aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac
     5641 gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag
     5701 atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg
     5761 tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca
     5821 gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac cacttcaaga
     5881 actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca
     5941 gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc
     6001 agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca
     6061 ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa
     6121 aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc
     6181 cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc
     6241 gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaa
//
