LOCUS       Exported                6351 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  AAV-mediated expression of Jaws under the human synapsin promoter.
ACCESSION   .
VERSION     .
KEYWORDS    pAAV-hsyn-Jaws-KGC-GFP-ER2
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6351)
  AUTHORS   Chuong AS, Miri ML, Busskamp V, Matthews GA, Acker LC, Sorensen AT, 
            Young A, Klapoetke NC, Henninger MA, Kodandaramaiah SB, Ogawa M, 
            Ramanlal SB, Bandler RC, Allen BD, Forest CR, Chow BY, Han X, Lin Y,
            Tye KM, Roska B, Cardin JA, Boyden ES
  TITLE     Noninvasive optical inhibition with a red-shifted microbial 
            rhodopsin.
  JOURNAL   Nat Neurosci. 2014 Aug;17(8):1123-9. doi: 10.1038/nn.3752. Epub 2014
            Jul 6.
  PUBMED    24997763
REFERENCE   2  (bases 1 to 6351)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6351
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     repeat_region   1..130
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     promoter        341..788
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     CDS             1722..2435
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(1764..1785)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2025..2044)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2372..2393
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    2484..3072
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2537..2557)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2955..2966)
                     /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   3614..3754
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3614..3743
                     /label=AAV2 ITR
     rep_origin      3829..4284
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3916..3935)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4126..4147
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4301..4320)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4420..4442
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4480..4498)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4566..4670
                     /gene="bla"
                     /label=AmpR promoter
     CDS             4671..5531
                     /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(4889..4908)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5702..6290
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6191..6210
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcgtcg ggcgaccttt
       61 ggtcgcccgg cctcagtgag cgagcgagcg cgcagagagg gagtggccaa ctccatcact
      121 aggggttcct gcggccgcac gcgttgcaaa gatggataaa gttttaaaca gagaggaatc
      181 tttgcagcta atggaccttc taggtcttga aaggagtggg aattggctcc ggtgcccgtc
      241 agtgggcaga gcgcacatcg cccacagtcc ccgagaagtt ggggggaggg gtcggcagca
      301 aatggttaat taatctagac tgcagagggc cctgcgtatg agtgcaagtg ggttttagga
      361 ccaggatgag gcggggtggg ggtgcctacc tgacgaccga ccccgaccca ctggacaagc
      421 acccaacccc cattccccaa attgcgcatc ccctatcaga gagggggagg ggaaacagga
      481 tgcggcgagg cgcgtgcgca ctgccagctt cagcaccgcg gacagtgcct tcgcccccgc
      541 ctggcggcgc gcgccaccgc cgcctcagca ctgaaggcgc gctgacgtca ctcgccggtc
      601 ccccgcaaac tccccttccc ggccaccttg gtcgcgtccg cgccgccgcc ggcccagccg
      661 gaccgcacca cgcgaggcgc gagatagggg ggcacgggcg cgaccatctg cgctgcggcg
      721 ccggcgactc agcgctgcct cagtctgcgg tgggcagcgg aggagtcgtg tcgtgcctga
      781 gagcgcagtc gagaggatcc gccaccatga ccgccgtgag caccacagcc actaccgtgc
      841 tgcaggccac acagagcgac gtgctgcagg agatccagtc caacttcctg ctgaatagct
      901 ccatctgggt gaacattgct ctggccggag tggtcatcct gctgtttgtg gccatgggga
      961 gggatctgga atcccctaga gctaagctga tctgggtggc cacaatgctg gtgccactgg
     1021 tgtctatttc tagttacgct ggactggcca gtgggctgac tgtgggcttc ctgcagatgc
     1081 cacctggaca cgctctggcc ggacaggagg tgctgagccc atggggccgg tatctgacat
     1141 ggactttctc cactcccatg atcctgctgg ctctgggact gctggccgac accgatattg
     1201 ccagcctgtt caccgccatc acaatggaca ttggcatgtg cgtgacagga ctggccgctg
     1261 ccctgatcac tagctcccat ctgctgcgct gggtgttcta cggaatttct tgtgctttct
     1321 ttgtggccgt gctgtatgtg ctgctggtgc agtggccagc tgatgctgag gctgctggga
     1381 ccagtgaaat ctttggcact ctgaggattc tgaccgtggt gctgtggctg gggtacccta
     1441 tcctgttcgc tctgggctct gagggagtgg ccctgctgag tgtgggagtg accagctggg
     1501 gatactccgg actggacatc ctggctaaat acgtgttcgc ctttctgctg ctgagatggg
     1561 tggctgccaa tgaaggcaca gtgtctggga gtggaatggg aatcgggtcc ggaggagctg
     1621 ctccagccga cgatcgaccg gtagtaaaat ccagaattac ttctgaaggg gagtatatcc
     1681 ctctggatca aatagacatc aatgttgcac ctgcaggggc agtgagcaag ggcgaggagc
     1741 tgttcaccgg ggtggtgccc atcctggtcg agctggacgg cgacgtaaac ggccacaagt
     1801 tcagcgtgtc cggcgagggc gagggcgatg ccacctacgg caagctgacc ctgaagttca
     1861 tttgcaccac cggcaagctg cccgtgccct ggcccaccct cgtgaccacc ctgacctacg
     1921 gcgtgcagtg cttcagccgc taccccgacc acatgaagca gcacgacttc ttcaagtccg
     1981 ccatgcccga aggctacgtc caggagcgca ccatcttctt caaggacgac ggcaactaca
     2041 agacccgcgc cgaggtgaag ttcgagggcg acaccctggt gaaccgcatc gagctgaagg
     2101 gcatcgactt caaggaggac ggcaacatcc tggggcacaa gctggagtac aactacaaca
     2161 gccacaacgt ctatatcatg gccgacaagc agaagaacgg catcaaggtg aacttcaaga
     2221 tccgccacaa catcgaggac ggcagcgtgc agctcgccga ccactaccag cagaacaccc
     2281 ccatcggcga cggccccgtg ctgctgcccg acaaccacta cctgagcacc cagtccgccc
     2341 tgagcaaaga ccccaacgag aagcgcgatc acatggtcct gctggagttc gtgaccgccg
     2401 ccgggatcac tctcggcatg gacgagctgt acaagttctg ctacgagaat gaagtgtaag
     2461 aattccgtac aaagcttatc gataatcaac ctctggatta caaaatttgt gaaagattga
     2521 ctggtattct taactatgtt gctcctttta cgctatgtgg atacgctgct ttaatgcctt
     2581 tgtatcatgc tattgcttcc cgtatggctt tcattttctc ctccttgtat aaatcctggt
     2641 tgctgtctct ttatgaggag ttgtggcccg ttgtcaggca acgtggcgtg gtgtgcactg
     2701 tgtttgctga cgcaaccccc actggttggg gcattgccac cacctgtcag ctcctttccg
     2761 ggactttcgc tttccccctc cctattgcca cggcggaact catcgccgcc tgccttgccc
     2821 gctgctggac aggggctcgg ctgttgggca ctgacaattc cgtggtgttg tcggggaaat
     2881 catcgtcctt tccttggctg ctcgcctatg ttgccacctg gattctgcgc gggacgtcct
     2941 tctgctacgt cccttcggcc ctcaatccag cggaccttcc ttcccgcggc ctgctgccgg
     3001 ctctgcggcc tcttccgcgt cttcgccttc gccctcagac gagtcggatc tccctttggg
     3061 ccgcctcccc gcatcgatac cggctcgaga 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 cgagctgatt ttgtaggtaa ccacgtgcgg
     3601 accgagcggc cgcaggaacc cctagtgatg gagttggcca ctccctctct gcgcgctcgc
     3661 tcgctcactg aggccgggcg accaaaggtc gcccgacgcc cgggctttgc ccgggcggcc
     3721 tcagtgagcg agcgagcgcg cagctgcctg caggggcgcc tgatgcggta ttttctcctt
     3781 acgcatctgt gcggtatttc acaccgcata cgtcaaagca accatagtac gcgccctgta
     3841 gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca
     3901 gcgccttagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg ttcgccggct
     3961 ttccccgtca agctctaaat cgggggctcc ctttagggtt ccgatttagt gctttacggc
     4021 acctcgaccc caaaaaactt gatttgggtg atggttcacg tagtgggcca tcgccctgat
     4081 agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga ctcttgttcc
     4141 aaactggaac aacactcaac tctatctcgg gctattcttt tgatttataa gggattttgc
     4201 cgatttcggt ctattggtta aaaaatgagc tgatttaaca aaaatttaac gcgaatttta
     4261 acaaaatatt aacgtttaca attttatggt gcactctcag tacaatctgc tctgatgccg
     4321 catagttaag ccagccccga cacccgccaa cacccgctga cgcgccctga cgggcttgtc
     4381 tgctcccggc atccgcttac agacaagctg tgaccgtctc cgggagctgc atgtgtcaga
     4441 ggttttcacc gtcatcaccg aaacgcgcga gacgaaaggg cctcgtgata cgcctatttt
     4501 tataggttaa tgtcatgata ataatggttt cttagacgtc aggtggcact tttcggggaa
     4561 atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca
     4621 tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt atgagtattc
     4681 aacatttccg tgtcgccctt attccctttt ttgcggcatt ttgccttcct gtttttgctc
     4741 acccagaaac gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt
     4801 acatcgaact ggatctcaac agcggtaaga tccttgagag ttttcgcccc gaagaacgtt
     4861 ttccaatgat gagcactttt aaagttctgc tatgtggcgc ggtattatcc cgtattgacg
     4921 ccgggcaaga gcaactcggt cgccgcatac actattctca gaatgacttg gttgagtact
     4981 caccagtcac agaaaagcat cttacggatg gcatgacagt aagagaatta tgcagtgctg
     5041 ccataaccat gagtgataac actgcggcca acttacttct gacaacgatc ggaggaccga
     5101 aggagctaac cgcttttttg cacaacatgg gggatcatgt aactcgcctt gatcgttggg
     5161 aaccggagct gaatgaagcc ataccaaacg acgagcgtga caccacgatg cctgtagcaa
     5221 tggcaacaac gttgcgcaaa ctattaactg gcgaactact tactctagct tcccggcaac
     5281 aattaataga ctggatggag gcggataaag ttgcaggacc acttctgcgc tcggcccttc
     5341 cggctggctg gtttattgct gataaatctg gagccggtga gcgtgggtct cgcggtatca
     5401 ttgcagcact ggggccagat ggtaagccct cccgtatcgt agttatctac acgacgggga
     5461 gtcaggcaac tatggatgaa cgaaatagac agatcgctga gataggtgcc tcactgatta
     5521 agcattggta actgtcagac caagtttact catatatact ttagattgat ttaaaacttc
     5581 atttttaatt taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc
     5641 cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt
     5701 cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac
     5761 cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct
     5821 tcagcagagc gcagatacca aatactgttc ttctagtgta gccgtagtta ggccaccact
     5881 tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg
     5941 ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata
     6001 aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga
     6061 cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag
     6121 ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg
     6181 agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac
     6241 ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca
     6301 acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg t
//
