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"
     primer_bind     178..197
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     437..458
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     promoter        complement(1434..1881)
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     repeat_region   2092..2221
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     rep_origin      complement(2283..2871)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2363..2382)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(3042..3902)
                     /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     3665..3684
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3903..4007)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4075..4093
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4131..4153)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4253..4272
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(4289..4744)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4426..4447)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4638..4657
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     repeat_region   4819..4959
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4830..4959
                     /label=AAV2 ITR
     polyA_signal    5005..5481
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     5263..5282
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     misc_feature    5501..6089
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     CDS             5607..5618
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     6016..6036
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(6138..500)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(6180..6201)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
ORIGIN
        1 atcttgaagt tcaccttgat gccgttcttc tgcttgtcgg ccatgatata gacgttgtgg
       61 ctgttgtagt tgtactccag cttgtgcccc aggatgttgc cgtcctcctt gaagtcgatg
      121 cccttcagct cgatgcggtt caccagggtg tcgccctcga acttcacctc ggcgcgggtc
      181 ttgtagttgc cgtcgtcctt gaagaagatg gtgcgctcct ggacgtagcc ttcgggcatg
      241 gcggacttga agaagtcgtg ctgcttcatg tggtcggggt agcggctgaa gcactgcacg
      301 ccgtaggtca gggtggtcac gagggtgggc cagggcacgg gcagcttgcc ggtggtgcaa
      361 atgaacttca gggtcagctt gccgtaggtg gcatcgccct cgccctcgcc ggacacgctg
      421 aacttgtggc cgtttacgtc gccgtccagc tcgaccagga tgggcaccac cccggtgaac
      481 agctcctcgc ccttgctcac tgcccctgca ggtgcaacat tgatgtctat ttgatccaga
      541 gggatatact ccccttcaga agtaattctg gattttacta ccggtcgatc gtcggctgga
      601 gcagctcctc cggacccgat tcccattcca ctcccagaca ctgtgccttc attggcagcc
      661 acccatctca gcagcagaaa ggcgaacacg tatttagcca ggatgtccag tccggagtat
      721 ccccagctgg tcactcccac actcagcagg gccactccct cagagcccag agcgaacagg
      781 atagggtacc ccagccacag caccacggtc agaatcctca gagtgccaaa gatttcactg
      841 gtcccagcag cctcagcatc agctggccac tgcaccagca gcacatacag cacggccaca
      901 aagaaagcac aagaaattcc gtagaacacc cagcgcagca gatgggagct agtgatcagg
      961 gcagcggcca gtcctgtcac gcacatgcca atgtccattg tgatggcggt gaacaggctg
     1021 gcaatatcgg tgtcggccag cagtcccaga gccagcagga tcatgggagt ggagaaagtc
     1081 catgtcagat accggcccca tgggctcagc acctcctgtc cggccagagc gtgtccaggt
     1141 ggcatctgca ggaagcccac agtcagccca ctggccagtc cagcgtaact agaaatagac
     1201 accagtggca ccagcattgt ggccacccag atcagcttag ctctagggga ttccagatcc
     1261 ctccccatgg ccacaaacag caggatgacc actccggcca gagcaatgtt cacccagatg
     1321 gagctattca gcaggaagtt ggactggatc tcctgcagca cgtcgctctg tgtggcctgc
     1381 agcacggtag tggctgtggt gctcacggcg gtcatggtgg cggatcctct cgactgcgct
     1441 ctcaggcacg acacgactcc tccgctgccc accgcagact gaggcagcgc tgagtcgccg
     1501 gcgccgcagc gcagatggtc gcgcccgtgc ccccctatct cgcgcctcgc gtggtgcggt
     1561 ccggctgggc cggcggcggc gcggacgcga ccaaggtggc cgggaagggg agtttgcggg
     1621 ggaccggcga gtgacgtcag cgcgccttca gtgctgaggc ggcggtggcg cgcgccgcca
     1681 ggcgggggcg aaggcactgt ccgcggtgct gaagctggca gtgcgcacgc gcctcgccgc
     1741 atcctgtttc ccctccccct ctctgatagg ggatgcgcaa tttggggaat gggggttggg
     1801 tgcttgtcca gtgggtcggg gtcggtcgtc aggtaggcac ccccaccccg cctcatcctg
     1861 gtcctaaaac ccacttgcac tcatacgcag ggccctctgc agtctagatt aattaaccat
     1921 ttgctgccga cccctccccc caacttctcg gggactgtgg gcgatgtgcg ctctgcccac
     1981 tgacgggcac cggagccaat tcccactcct ttcaagacct agaaggtcca ttagctgcaa
     2041 agattcctct ctgtttaaaa ctttatccat ctttgcaacg cgtgcggccg caggaacccc
     2101 tagtgatgga gttggccact ccctctctgc gcgctcgctc gctcactgag gccgggcgac
     2161 caaaggtcgc ccgacgcccg ggcggcctca gtgagcgagc gagcgcgcag ctgcctgcag
     2221 gacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg
     2281 tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg
     2341 tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg
     2401 cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga
     2461 agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc
     2521 tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt
     2581 aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact
     2641 ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg
     2701 cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt
     2761 accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt
     2821 ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct
     2881 ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg
     2941 gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt
     3001 aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt
     3061 gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc
     3121 gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg
     3181 cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc
     3241 gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg
     3301 gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca
     3361 ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga
     3421 tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct
     3481 ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg
     3541 cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca
     3601 accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata
     3661 cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct
     3721 tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact
     3781 cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa
     3841 acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc
     3901 atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga
     3961 tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga
     4021 aaagtgccac ctgacgtcta agaaaccatt attatcatga cattaaccta taaaaatagg
     4081 cgtatcacga ggccctttcg tctcgcgcgt ttcggtgatg acggtgaaaa cctctgacac
     4141 atgcagctcc cggagacggt cacagcttgt ctgtaagcgg atgccgggag cagacaagcc
     4201 cgtcagggcg cgtcagcggg tgttggcggg tgtcggggct ggcttaacta tgcggcatca
     4261 gagcagattg tactgagagt gcaccataaa attgtaaacg ttaatatttt gttaaaattc
     4321 gcgttaaatt tttgttaaat cagctcattt tttaaccaat agaccgaaat cggcaaaatc
     4381 ccttataaat caaaagaata gcccgagata gagttgagtg ttgttccagt ttggaacaag
     4441 agtccactat taaagaacgt ggactccaac gtcaaagggc gaaaaaccgt ctatcagggc
     4501 gatggcccac tacgtgaacc atcacccaaa tcaagttttt tggggtcgag gtgccgtaaa
     4561 gcactaaatc ggaaccctaa agggagcccc cgatttagag cttgacgggg aaagccggcg
     4621 aacgtggcga gaaaggaagg gaagaaagcg aaaggagcgg gcgctaaggc gctggcaagt
     4681 gtagcggtca cgctgcgcgt aaccaccaca cccgccgcgc ttaatgcgcc gctacagggc
     4741 gcgtactatg gttgctttga cgtatgcggt gtgaaatacc gcacagatgc gtaaggagaa
     4801 aataccgcat caggcgcccc tgcaggcagc tgcgcgctcg ctcgctcact gaggccgccc
     4861 gggcaaagcc cgggcgtcgg gcgacctttg gtcgcccggc ctcagtgagc gagcgagcgc
     4921 gcagagaggg agtggccaac tccatcacta ggggttcctg cggccgctcg gtccgcacgt
     4981 ggttacctac aaaatcagct cgagaaggac agggaaggga gcagtggttc acgcctgtaa
     5041 tcccagcaat ttgggaggcc aaggtgggta gatcacctga gattaggagt tggagaccag
     5101 cctggccaat atggtgaaac cccgtctcta ccaaaaaaac aaaaattagc tgagcctggt
     5161 catgcatgcc tggaatccca acaactcggg aggctgaggc aggagaatcg cttgaaccca
     5221 ggaggcggag attgcagtga gccaagattg tgccactgca ctccagcttg gttcccaata
     5281 gaccccgcag gccctacagg ttgtcttccc aacttgcccc ttgctccata ccacccccct
     5341 ccaccccata atattataga aggacaccta gtcagacaaa atgatgcaac ttaattttat
     5401 taggacaagg ctggtgggca ctggagtggc aacttccagg gccaggagag gcactgggga
     5461 ggggtcacag ggatgccacc cgtctcgagc cggtatcgat gcggggaggc ggcccaaagg
     5521 gagatccgac tcgtctgagg gcgaaggcga agacgcggaa gaggccgcag agccggcagc
     5581 aggccgcggg aaggaaggtc cgctggattg agggccgaag ggacgtagca gaaggacgtc
     5641 ccgcgcagaa tccaggtggc aacataggcg agcagccaag gaaaggacga tgatttcccc
     5701 gacaacacca cggaattgtc agtgcccaac agccgagccc ctgtccagca gcgggcaagg
     5761 caggcggcga tgagttccgc cgtggcaata gggaggggga aagcgaaagt cccggaaagg
     5821 agctgacagg tggtggcaat gccccaacca gtgggggttg cgtcagcaaa cacagtgcac
     5881 accacgccac gttgcctgac aacgggccac aactcctcat aaagagacag caaccaggat
     5941 ttatacaagg aggagaaaat gaaagccata cgggaagcaa tagcatgata caaaggcatt
     6001 aaagcagcgt atccacatag cgtaaaagga gcaacatagt taagaatacc agtcaatctt
     6061 tcacaaattt tgtaatccag aggttgatta tcgataagct ttgtacggaa ttcttacact
     6121 tcattctcgt agcagaactt gtacagctcg tccatgccga gagtgatccc ggcggcggtc
     6181 acgaactcca gcaggaccat gtgatcgcgc ttctcgttgg ggtctttgct cagggcggac
     6241 tgggtgctca ggtagtggtt gtcgggcagc agcacggggc cgtcgccgat gggggtgttc
     6301 tgctggtagt ggtcggcgag ctgcacgctg ccgtcctcga tgttgtggcg g
//
