LOCUS       Exported                7934 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  An AAV vector that expresses GFP under the control of the L7-6 
            promoter. The L7-6 promoter is the short size (0.8-kb) and has 
            highly specificity to Purkinje cells..
ACCESSION   .
VERSION     .
KEYWORDS    pAAV/L7-6-GFP-WPRE
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7934)
  AUTHORS   Nitta K, Matsuzaki Y, Konno A, Hirai H
  TITLE     Minimal Purkinje Cell-Specific PCP2/L7 Promoter Virally Available 
            for Rodents and Non-human Primates.
  JOURNAL   Mol Ther Methods Clin Dev. 2017 Jul 27;6:159-170. doi: 
            10.1016/j.omtm.2017.07.006. eCollection 2017 Sep 15.
  PUBMED    28828391
REFERENCE   2  (bases 1 to 7934)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7934
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     regulatory      970..979
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             976..1695
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             976..1692
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1021..1042)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1282..1301)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1629..1650
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    1711..2299
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(1764..1784)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2182..2193)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    2426..2820
                     /label=-beta-globin poly(A)
                     /note="human beta-globin polyadenylation signal"
     repeat_region   4292..4421
                     /label=AAV2 ITR
     rep_origin      4792..5305
                     /direction=RIGHT
                     /label=M13 ori
                     /note="M13 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4879..4898)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5089..5110
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5787..5806)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5906..5928
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5966..5984)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6052..6156
                     /gene="bla"
                     /label=AmpR promoter
     CDS             6157..7017
                     /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(6375..6394)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7188..7776
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7677..7696
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7930..13
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 gtcagtgagc gagcgagcgc gcagagaggg agtggccaac tccatcacta ggggttcctt
       61 gtagttaatg attaacccgc catgctactt atctacgtag ccatgctcta gactcgaggg
      121 ttccaccctc atgttggttg atcttcaaca ttgtcctgac ttcttcccac ttgacattcc
      181 tcagggtcct gtgatcatgg ctgggtctag tgaggttcaa acctgcactg ccctacccac
      241 acccacaccc agctcagcgt cagtcaggat caacagttac ctagagatca tctttctggg
      301 gcttaagcat tggtgggagc agatgggata tgagttgggg atttggggat gggggaagat
      361 atctgctccc cctcccccta caccctagcc ttttaaaagg ccttctcagg tcagagacca
      421 ggagaaaagt ataggagaga tacacaatgg accaggaaga agaaaaggga gagggaggct
      481 cagaccttct agacaaggta agagggctct ggctgactcc accatccgct tcttgaggtc
      541 tcggcacctg taattgacaa gattaattca tttatagggc atctaattag caagtaagtc
      601 tctggagtcc cctgacccag ttactataac acacaggggg tataggtagg ggagtataag
      661 agcccctcct cagggcaaat gaatggattc ttagtactgt cccccaagag atagtaggta
      721 ctaggattta ggggcacttc tgagccccat ttccctggta agtgtcccaa ccccccaaat
      781 caacccaagc ctggtctcaa tctaggacag tggtagaatg ctgtccctag agtcagtacc
      841 atgtgaaatt gtgctgcagg caggggcccc aggctgggag gtgggggttg ggggagtcag
      901 gggcaggtca gggaaggaga ctcaggtttc atttagagaa attctgcaga cccgtgagga
      961 ctaccggtcg ccaccatggt gagcaagggc gaggagctgt tcaccggggt ggtgcccatc
     1021 ctggtcgagc tggacggcga cgtaaacggc cacaagttca gcgtgtccgg cgagggcgag
     1081 ggcgatgcca cctacggcaa gctgaccctg aagttcatct gcaccaccgg caagctgccc
     1141 gtgccctggc ccaccctcgt gaccaccctg acctacggcg tgcagtgctt cagccgctac
     1201 cccgaccaca tgaagcagca cgacttcttc aagtccgcca tgcccgaagg ctacgtccag
     1261 gagcgcacca tcttcttcaa ggacgacggc aactacaaga cccgcgccga ggtgaagttc
     1321 gagggcgaca ccctggtgaa ccgcatcgag ctgaagggca tcgacttcaa ggaggacggc
     1381 aacatcctgg ggcacaagct ggagtacaac tacaacagcc acaacgtcta tatcatggcc
     1441 gacaagcaga agaacggcat caaggtgaac ttcaagatcc gccacaacat cgaggacggc
     1501 agcgtgcagc tcgccgacca ctaccagcag aacaccccca tcggcgacgg ccccgtgctg
     1561 ctgcccgaca accactacct gagcacccag tccgccctga gcaaagaccc caacgagaag
     1621 cgcgatcaca tggtcctgct ggagttcgtg accgccgccg ggatcactct cggcatggac
     1681 gagctgtaca agtaaagcgg ccgcatcgat aatcaacctc tggattacaa aatttgtgaa
     1741 agattgactg gtattcttaa ctatgttgct ccttttacgc tatgtggata cgctgcttta
     1801 atgcctttgt atcatgctat tgcttcccgt atggctttca ttttctcctc cttgtataaa
     1861 tcctggttgc tgtctcttta tgaggagttg tggcccgttg tcaggcaacg tggcgtggtg
     1921 tgcactgtgt ttgctgacgc aacccccact ggttggggca ttgccaccac ctgtcagctc
     1981 ctttccggga ctttcgcttt ccccctccct attgccacgg cggaactcat cgccgcctgc
     2041 cttgcccgct gctggacagg ggctcggctg ttgggcactg acaattccgt ggtgttgtcg
     2101 gggaagctga cgtcctttcc atggctgctc gcctgtgttg ccacctggat tctgcgcggg
     2161 acgtccttct gctacgtccc ttcggccctc aatccagcgg accttccttc ccgcggcctg
     2221 ctgccggctc tgcggcctct tccgcgtctt cgccttcgcc ctcagacgag tcggatctcc
     2281 ctttgggccg cctccccgca tcgatgccgt atacggtacc cggggatcct ctagtagagt
     2341 cgaggaattc accccaccag tgcaggctgc ctatcagaaa gtggtggctg gtgtggctaa
     2401 tgccctggcc cacaagtatc actaagctcg ctttcttgct gtccaatttc tattaaaggt
     2461 tcctttgttc cctaagtcca actactaaac tgggggatat tatgaagggc cttgagcatc
     2521 tggattctgc ctaataaaaa acatttattt tcattgcaat gatgtattta aattatttct
     2581 gaatatttta ctaaaaaggg aatgtgggag gtcagtgcat ttaaaacata aagaaatgaa
     2641 gagctagttc aaaccttggg aaaatacact atatcttaaa ctccatgaaa gaaggtgagg
     2701 ctgcaaacag ctaatgcaca ttggcaacag cccctgatgc ctatgcctta ttcatccctc
     2761 agaaaaggat tcaagtagag gcttgatttg gaggttaaag ttttgctatg ctgtatttta
     2821 cattacttat tgttttagct gtcctcatga atgtcttttc actacccatt tgcttatcct
     2881 gcatctctca gccttgactc cactcagttc tcttgcttag agataccacc tttcccctga
     2941 agtgttcctt ccatgtttta cggcgagatg gtttctcctc gcctggccac tcagccttag
     3001 ttgtctctgt tgtcttatag aggtctactt gaagaaggaa aaacaggggg catggtttga
     3061 ctgtcctgtg agcccttctt ccctgcctcc cccactcaca gtgacccgga atctgcagtg
     3121 ctagtctccc ggaactatca ctctttcaca gtctgctttg gaaggactgg gcttagtatg
     3181 aaaagttagg actgagaaga atttgaaagg gggctttttg tagcttgata ttcactactg
     3241 tcttattacc ctatcatagg cccaccccaa atggaagtcc cattcttcct caggatgttt
     3301 aagattagca ttcaggaaga gatcagaggt ctgctggctc ccttatcatg tcccttatgg
     3361 tgcttctggc tctgcagtta ttagcatagt gttaccatca accaccttaa cttcattttt
     3421 cttattcaat acctaggtag gtagatgcta gattctggaa ataaaatatg agtctcaagt
     3481 ggtccttgtc ctctctccca gtcaaattct gaatctagtt ggcaagattc tgaaatcaag
     3541 gcatataatc agtaataagt gatgatagaa gggtatatag aagaatttta ttatatgaga
     3601 gggtgaaacc ctcaaaatga aatgaaatca gacccttgtc ttacaccata aacaaaaata
     3661 aatttgaatg ggttaaagaa ttaaactaag acctaaaacc ataaaaattt ttaaagaaat
     3721 caaaagaaga aaattctaat attcatgttg cagccgtttt ttgaatttga tatgagaagc
     3781 aaaggcaaca aaaggaaaaa taaagaagtg aggctacatc aaactaaaaa atttccacac
     3841 aaaaaagaaa acaatgaaca aatgaaaggt gaaccatgaa atggcatatt tgcaaaccaa
     3901 atatttctta aatattttgg ttaatatcca aaatatataa gaaacacaga tgattcaata
     3961 acaaacaaaa aattaaaaat aggaaaataa aaaaattaaa aagaagaaaa tcctgccatt
     4021 tatggcagaa ttgatgaacc tggaggatgt aaaactaaga aaaataagcc tgacacaaaa
     4081 agacaaatac tacacaacct tgctcatatg tgaaacataa aaaagtcact ctcatggaaa
     4141 cagacagtag aggtatggtt tccaggggtt gggggtggga gaatcaggaa actattactc
     4201 aaagggtata aaatttcagt tatgtgggat gaataaattc tagagcatgg ctacgtagat
     4261 aagtagcatg gcgggttaat cattaactac aaggaacccc tagtgatgga gttggccact
     4321 ccctctctgc gcgctcgctc gctcactgag gccgggcgac caaaggtcgc ccgacgcccg
     4381 ggctttgccc gggcggcctc agtgagcgag cgagcgcgca gctggcgtaa tagcgaagag
     4441 gcccgcaccg atcgcccttc ccaacagttg cgcagcctga atggcgaatg gcgattccgt
     4501 tgcaatggct ggcggtaata ttgttctgga tattaccagc aaggccgata gtttgagttc
     4561 ttctactcag gcaagtgatg ttattactaa tcaaagaagt attgcgacaa cggttaattt
     4621 gcgtgatgga cagactcttt tactcggtgg cctcactgat tataaaaaca cttctcagga
     4681 ttctggcgta ccgttcctgt ctaaaatccc tttaatcggc ctcctgttta gctcccgctc
     4741 tgattctaac gaggaaagca cgttatacgt gctcgtcaaa gcaaccatag tacgcgccct
     4801 gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg
     4861 ccagcgccct agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg
     4921 gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt agtgctttac
     4981 ggcacctcga ccccaaaaaa cttgattagg gtgatggttc acgtagtggg ccatcgccct
     5041 gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt ggactcttgt
     5101 tccaaactgg aacaacactc aaccctatct cggtctattc ttttgattta taagggattt
     5161 tgccgatttc ggcctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt
     5221 ttaacaaaat attaacgctt acaatttaaa tatttgctta tacaatcttc ctgtttttgg
     5281 ggcttttctg attatcaacc ggggtacata tgattgacat gctagtttta cgattaccgt
     5341 tcatcgattc tcttgtttgc tccagactct caggcaatga cctgatagcc tttgtagaga
     5401 cctctcaaaa atagctaccc tctccggcat gaatttatca gctagaacgg ttgaatatca
     5461 tattgatggt gatttgactg tctccggcct ttctcacccg tttgaatctt tacctacaca
     5521 ttactcaggc attgcattta aaatatatga gggttctaaa aatttttatc cttgcgttga
     5581 aataaaggct tctcccgcaa aagtattaca gggtcataat gtttttggta caaccgattt
     5641 agctttatgc tctgaggctt tattgcttaa ttttgctaat tctttgcctt gcctgtatga
     5701 tttattggat gttggaatcg cctgatgcgg tattttctcc ttacgcatct gtgcggtatt
     5761 tcacaccgca tatggtgcac tctcagtaca atctgctctg atgccgcata gttaagccag
     5821 ccccgacacc cgccaacacc cgctgacgcg ccctgacggg cttgtctgct cccggcatcc
     5881 gcttacagac aagctgtgac cgtctccggg agctgcatgt gtcagaggtt ttcaccgtca
     5941 tcaccgaaac gcgcgagacg aaagggcctc gtgatacgcc tatttttata ggttaatgtc
     6001 atgataataa tggtttctta gacgtcaggt ggcacttttc ggggaaatgt gcgcggaacc
     6061 cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc
     6121 tgataaatgc ttcaataata ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc
     6181 gcccttattc ccttttttgc ggcattttgc cttcctgttt ttgctcaccc agaaacgctg
     6241 gtgaaagtaa aagatgctga agatcagttg ggtgcacgag tgggttacat cgaactggat
     6301 ctcaacagcg gtaagatcct tgagagtttt cgccccgaag aacgttttcc aatgatgagc
     6361 acttttaaag ttctgctatg tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa
     6421 ctcggtcgcc gcatacacta ttctcagaat gacttggttg agtactcacc agtcacagaa
     6481 aagcatctta cggatggcat gacagtaaga gaattatgca gtgctgccat aaccatgagt
     6541 gataacactg cggccaactt acttctgaca acgatcggag gaccgaagga gctaaccgct
     6601 tttttgcaca acatggggga tcatgtaact cgccttgatc gttgggaacc ggagctgaat
     6661 gaagccatac caaacgacga gcgtgacacc acgatgcctg tagcaatggc aacaacgttg
     6721 cgcaaactat taactggcga actacttact ctagcttccc ggcaacaatt aatagactgg
     6781 atggaggcgg ataaagttgc aggaccactt ctgcgctcgg cccttccggc tggctggttt
     6841 attgctgata aatctggagc cggtgagcgt gggtctcgcg gtatcattgc agcactgggg
     6901 ccagatggta agccctcccg tatcgtagtt atctacacga cggggagtca ggcaactatg
     6961 gatgaacgaa atagacagat cgctgagata ggtgcctcac tgattaagca ttggtaactg
     7021 tcagaccaag tttactcata tatactttag attgatttaa aacttcattt ttaatttaaa
     7081 aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt
     7141 tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt
     7201 tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt
     7261 ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag
     7321 ataccaaata ctgttcttct agtgtagccg tagttaggcc accacttcaa gaactctgta
     7381 gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat
     7441 aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg
     7501 ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg
     7561 agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac
     7621 aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga
     7681 aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt
     7741 ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta
     7801 cggttcctgg ccttttgctg gccttttgct cacatgttct ttcctgcgtt atcccctgat
     7861 tctgtggata accgtattac cgcctttgag tgagctgata ccgctcgccg cagccgaacg
     7921 accgagcgca gcga
//
