LOCUS       sequence-466781-        6164 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pAAV transfer plasmid with enhanced Synapsin promoter driving 
            Flp-dependent expression of TVA950-eYFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6164)
  AUTHORS   Mendelsohn AI, Nikoobakht L, Bikoff JB, Costa RM
  TITLE     Segregated basal ganglia output pathways correspond to genetically 
            divergent neuronal subclasses.
  JOURNAL   bioRxiv [Preprint]. 2024 Sep 18:2024.08.28.610136. doi: 
            10.1101/2024.08.28.610136.
  PUBMED    39257765
REFERENCE   2  (bases 1 to 6164)
  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: "bioRxiv 
            [Preprint]. 2024 Sep 18:2024.08.28.610136. doi: 
            10.1101/2024.08.28.610136."
COMMENT     Sequence Label: pAAV-Esyn-FlpX-TVA950-EYFP-WPRE
FEATURES             Location/Qualifiers
     source          1..6164
                     /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..780
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     enhancer        855..1158
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     protein_bind    1575..1608
                     /label=FRT (minimal)
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="supports FLP-mediated excision but not integration 
                     (Turan and Bode, 2011)"
     protein_bind    1696..1729
                     /label=FRT3
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="supports FLP-mediated excision with other FRT3 sites
                     (Bosch et al., 2015)"
     CDS             complement(join(1746..2459,2460..2462,2463..2465))
                     /codon_start=1
                     /product="enhanced YFP"
                     /label=EYFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1791..1812)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     2140..2159
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2399..2420
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      2462..2471
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     regulatory      2945..2954
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    complement(2997..3030)
                     /label=FRT (minimal)
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="supports FLP-mediated excision but not integration 
                     (Turan and Bode, 2011)"
     protein_bind    complement(3118..3151)
                     /label=FRT3
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="supports FLP-mediated excision with other FRT3 sites
                     (Bosch et al., 2015)"
     misc_feature    3190..3778
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3243..3263)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3661..3672)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(3805..3822)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3811..4035
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   4077..4217
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4077..4206
                     /label=AAV2 ITR
     rep_origin      4292..4747
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4379..4398)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4589..4610
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4764..4783)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4883..4905
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4943..4961)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5029..5133
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5134..5202,5203..5994)
                     /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(5352..5371)
                     /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 ccgggcgtcg ggcgaccttt ggtcgcccgg
      721 cctcagtgag cgagcgagcg cgcagagagg gagtggccaa ctccatcact aggggttcct
      781 gcggccgcac gcgttagtta ttaatagtaa tcaattacgg ggtcattagt tcatagccca
      841 tatatggagt tccgcgttac ataacttacg gtaaatggcc cgcctggctg accgcccaac
      901 gacccccgcc cattgacgtc aataatgacg tatgttccca tagtaacgcc aatagggact
      961 ttccattgac gtcaatgggt ggagtattta cggtaaactg cccacttggc agtacatcaa
     1021 gtgtatcata tgccaagtac gccccctatt gacgtcaatg acggtaaatg gcccgcctgg
     1081 cattatgccc agtacatgac cttatgggac tttcctactt ggcagtacat ctacgtatta
     1141 gtcatcgcta ttaccatgga gatctctgca gagggccctg cgtatgagtg caagtgggtt
     1201 ttaggaccag gatgaggcgg ggtgggggtg cctacctgac gaccgacccc gacccactgg
     1261 acaagcaccc aacccccatt ccccaaattg cgcatcccct atcagagagg gggaggggaa
     1321 acaggatgcg gcgaggcgcg tgcgcactgc cagcttcagc accgcggaca gtgccttcgc
     1381 ccccgcctgg cggcgcgcgc caccgccgcc tcagcactga aggcgcgctg acgtcactcg
     1441 ccggtccccc gcaaactccc cttcccggcc accttggtcg cgtccgcgcc gccgccggcc
     1501 cagccggacc gcaccacgcg aggcgcgaga taggggggca cgggcgcgac catctgcgct
     1561 gcggcgggat ccccgaagtt cctattctct agaaagtata ggaacttcat caaaatagga
     1621 agaccaatgc ttcaccatcg acccgaattg ccaagcatca ccatcgaccc tcttgaccgt
     1681 ttcagattga cccgtgaagt tcctattctt caaatagtat aggaacttcg aattcctgca
     1741 ggtttttact tgtacagctc gtccatgccg agagtgatcc cggcggcggt cacgaactcc
     1801 agcaggacca tgtgatcgcg cttctcgttg gggtctttgc tcagggcgga ctggtagctc
     1861 aggtagtggt tgtcgggcag cagcacgggg ccgtcgccga tgggggtgtt ctgctggtag
     1921 tggtcggcga gctgcacgct gccgtcctcg atgttgtggc ggatcttgaa gttcaccttg
     1981 atgccgttct tctgcttgtc ggccatgata tagacgttgt ggctgttgta gttgtactcc
     2041 agcttgtgcc ccaggatgtt gccgtcctcc ttgaagtcga tgcccttcag ctcgatgcgg
     2101 ttcaccaggg tgtcgccctc gaacttcacc tcggcgcggg tcttgtagtt gccgtcgtcc
     2161 ttgaagaaga tggtgcgctc ctggacgtag ccttcgggca tggcggactt gaagaagtcg
     2221 tgctgcttca tgtggtcggg gtagcgggcg aagcactgca ggccgtagcc gaaggtggtc
     2281 acgagggtgg gccagggcac gggcagcttg ccggtggtgc agatgaactt cagggtcagc
     2341 ttgccgtagg tggcatcgcc ctcgccctcg ccggacacgc tgaacttgtg gccgtttacg
     2401 tcgccgtcca gctcgaccag gatgggcacc accccggtga acagctcctc gcccttgctc
     2461 accatggtgg cggccgcgga gaacaagtct gcctggctct tgtcaggcac cagcagctcc
     2521 ttggatgcgc tttcaagact gaagatgtca gacctgcttt ttgctttgga cttcccccat
     2581 gcagcgatac cacccacagc taccaggcag cacaggagca ctgcagtgat cagcatccac
     2641 atgcggccgt gattcctggc tggcagagca cgtccaggag caggggcagt gggagcctct
     2701 gtgccgttgt ccgtgggcac cgcggggatc gcgctggtcc cgcagcccca ctcgtcccgc
     2761 ccgtcgtcgc agtcggggtg tccgtcgcac agccagtcct gcgggtaaca ctccccgtgg
     2821 gcaccgggcg gctccgagca gcggaactga ccgggggggc aacgggacaa agaaccgtta
     2881 ccggacccgt taccggtcac gttaccgggc agcagcagca gcagcagcgc gggcagcagc
     2941 cgcgccatgg tggcaaacga gctcactagt gcggccgcct cgagagatct gatatcgaag
     3001 ttcctatact ttctagagaa taggaacttc cattgggatt cttcctattt tgatccaagc
     3061 atcaccatcg accctctagt ccagatctca ccatcgaccc taactttgaa tccgggtgaa
     3121 gttcctatac tatttgaaga ataggaactt cgtccctcga agaggttcaa gctttaacag
     3181 cttatcgata atcaacctct ggattacaaa atttgtgaaa gattgactgg tattcttaac
     3241 tatgttgctc cttttacgct atgtggatac gctgctttaa tgcctttgta tcatgctatt
     3301 gcttcccgta tggctttcat tttctcctcc ttgtataaat cctggttgct gtctctttat
     3361 gaggagttgt ggcccgttgt caggcaacgt ggcgtggtgt gcactgtgtt tgctgacgca
     3421 acccccactg gttggggcat tgccaccacc tgtcagctcc tttccgggac tttcgctttc
     3481 cccctcccta ttgccacggc ggaactcatc gccgcctgcc ttgcccgctg ctggacaggg
     3541 gctcggctgt tgggcactga caattccgtg gtgttgtcgg ggaaatcatc gtcctttcct
     3601 tggctgctcg cctgtgttgc cacctggatt ctgcgcggga cgtccttctg ctacgtccct
     3661 tcggccctca atccagcgga ccttccttcc cgcggcctgc tgccggctct gcggcctctt
     3721 ccgcgtcttc gccttcgccc tcagacgagt cggatctccc tttgggccgc ctccccgcat
     3781 cgataccgtc gactcgctga tcagcctcga ctgtgccttc tagttgccag ccatctgttg
     3841 tttgcccctc ccccgtgcct tccttgaccc tggaaggtgc cactcccact gtcctttcct
     3901 aataaaatga ggaaattgca tcgcattgtc tgagtaggtg tcattctatt ctggggggtg
     3961 gggtggggca ggacagcaag ggggaggatt gggaagacaa tagcaggcat gctggggatg
     4021 cggtgggctc tatggcttct gaggcggaaa gaaccaggtg cggaccgagc ggccgcagga
     4081 acccctagtg atggagttgg ccactccctc tctgcgcgct cgctcgctca ctgaggccgg
     4141 gcgaccaaag gtcgcccgac gcccgggctt tgcccgggcg gcctcagtga gcgagcgagc
     4201 gcgcagctgc ctgcaggggc gcctgatgcg gtattttctc cttacgcatc tgtgcggtat
     4261 ttcacaccgc atacgtcaaa gcaaccatag tacgcgccct gtagcggcgc attaagcgcg
     4321 gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgcctt agcgcccgct
     4381 cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta
     4441 aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa
     4501 cttgatttgg gtgatggttc acgtagtggg ccatcgccct gatagacggt ttttcgccct
     4561 ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg aacaacactc
     4621 aactctatct cgggctattc ttttgattta taagggattt tgccgatttc ggtctattgg
     4681 ttaaaaaatg agctgattta acaaaaattt aacgcgaatt ttaacaaaat attaacgttt
     4741 acaattttat ggtgcactct cagtacaatc tgctctgatg ccgcatagtt aagccagccc
     4801 cgacacccgc caacacccgc tgacgcgccc tgacgggctt gtctgctccc ggcatccgct
     4861 tacagacaag ctgtgaccgt ctccgggagc tgcatgtgtc agaggttttc accgtcatca
     4921 ccgaaacgcg cgagacgaaa gggcctcgtg atacgcctat ttttataggt taatgtcatg
     4981 ataataatgg tttcttagac gtcaggtggc acttttcggg gaaatgtgcg cggaacccct
     5041 atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga
     5101 taaatgcttc aataatattg aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc
     5161 cttattccct tttttgcggc attttgcctt cctgtttttg ctcacccaga aacgctggtg
     5221 aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg gttacatcga actggatctc
     5281 aacagcggta agatccttga gagttttcgc cccgaagaac gttttccaat gatgagcact
     5341 tttaaagttc tgctatgtgg cgcggtatta tcccgtattg acgccgggca agagcaactc
     5401 ggtcgccgca tacactattc tcagaatgac ttggttgagt actcaccagt cacagaaaag
     5461 catcttacgg atggcatgac agtaagagaa ttatgcagtg ctgccataac catgagtgat
     5521 aacactgcgg ccaacttact tctgacaacg atcggaggac cgaaggagct aaccgctttt
     5581 ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa
     5641 gccataccaa acgacgagcg tgacaccacg atgcctgtag caatggcaac aacgttgcgc
     5701 aaactattaa ctggcgaact acttactcta gcttcccggc aacaattaat agactggatg
     5761 gaggcggata aagttgcagg accacttctg cgctcggccc ttccggctgg ctggtttatt
     5821 gctgataaat ctggagccgg tgagcgtggg tctcgcggta tcattgcagc actggggcca
     5881 gatggtaagc cctcccgtat cgtagttatc tacacgacgg ggagtcaggc aactatggat
     5941 gaacgaaata gacagatcgc tgagataggt gcctcactga ttaagcattg gtaactgtca
     6001 gaccaagttt actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg
     6061 atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg
     6121 ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttc
//
