LOCUS       Exported                6557 bp ds-DNA     circular SYN 13-MAY-2021
DEFINITION  AAV-mediated expression of Jaws-KGC-tdTomato-ER2 under the Syn 
            promoter..
ACCESSION   .
VERSION     .
KEYWORDS    AAV-Syn-Jaws-KGC-tdTomato-ER2
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6557)
  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 6557)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 13, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6557
                     /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        164..611
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     misc_feature    2998..3586
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3051..3071)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3469..3480)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3594..3818
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     repeat_region   3820..3960
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3820..3949
                     /label=AAV2 ITR
     rep_origin      4035..4490
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4122..4141)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4332..4353
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4507..4526)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4626..4648
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4686..4704)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4772..4876
                     /gene="bla"
                     /label=AmpR promoter
     CDS             4877..5737
                     /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(5095..5114)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5908..6496
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6397..6416
                     /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 gtcgacgcta gactgcagag ggccctgcgt atgagtgcaa gtgggtttta
      181 ggaccaggat gaggcggggt gggggtgcct acctgacgac cgaccccgac ccactggaca
      241 agcacccaac ccccattccc caaattgcgc atcccctatc agagaggggg aggggaaaca
      301 ggatgcggcg aggcgcgtgc gcactgccag cttcagcacc gcggacagtg ccttcgcccc
      361 cgcctggcgg cgcgcgccac cgccgcctca gcactgaagg cgcgctgacg tcactcgccg
      421 gtcccccgca aactcccctt cccggccacc ttggtcgcgt ccgcgccgcc gccggcccag
      481 ccggaccgca ccacgcgagg cgcgagatag gggggcacgg gcgcgaccat ctgcgctgcg
      541 gcgccggcga ctcagcgctg cctcagtctg cggtgggcag cggaggagtc gtgtcgtgcc
      601 tgagagcgca gtcgaattcg ccgccaccat gaccgccgtg agcaccacag ccactaccgt
      661 gctgcaggcc acacagagcg acgtgctgca ggagatccag tccaacttcc tgctgaatag
      721 ctccatctgg gtgaacattg ctctggccgg agtggtcatc ctgctgtttg tggccatggg
      781 gagggatctg gaatccccta gagctaagct gatctgggtg gccacaatgc tggtgccact
      841 ggtgtctatt tctagttacg ctggactggc cagtgggctg actgtgggct tcctgcagat
      901 gccacctgga cacgctctgg ccggacagga ggtgctgagc ccatggggcc ggtatctgac
      961 atggactttc tccactccca tgatcctgct ggctctggga ctgctggccg acaccgatat
     1021 tgccagcctg ttcaccgcca tcacaatgga cattggcatg tgcgtgacag gactggccgc
     1081 tgccctgatc actagctccc atctgctgcg ctgggtgttc tacggaattt cttgtgcttt
     1141 ctttgtggcc gtgctgtatg tgctgctggt gcagtggcca gctgatgctg aggctgctgg
     1201 gaccagtgaa atctttggca ctctgaggat tctgaccgtg gtgctgtggc tggggtaccc
     1261 tatcctgttc gctctgggct ctgagggagt ggccctgctg agtgtgggag tgaccagctg
     1321 gggatactcc ggactggaca tcctggctaa atacgtgttc gcctttctgc tgctgagatg
     1381 ggtggctgcc aatgaaggca cagtgtctgg gagtggaatg ggaatcgggt ccggaggagc
     1441 tgctccagcc gacgatcgac cggtagtaaa atccagaatt acttctgaag gggagtatat
     1501 ccctctggat caaatagaca tcaatgttgc acctgcaggg gcagtgagta agggcgagga
     1561 agtgatcaaa gagttcatgc ggtttaaggt gagaatggaa ggaagcatga acggccacga
     1621 gttcgaaatt gagggagaag gagagggacg gccctacgag ggcacccaga cagccaagct
     1681 gaaagtgaca aagggcgggc ctctgccatt cgcttgggac atcctgagcc cacagtttat
     1741 gtacggctcc aaggcctatg tgaaacatcc agctgacatt cccgattata agaaactgag
     1801 cttccccgag gggtttaagt gggaaagagt gatgaacttc gaggacggag gcctggtgac
     1861 tgtgacccag gacagctccc tgcaggatgg gaccctgatc tacaaggtga aaatgagagg
     1921 gacaaatttt ccccctgatg gacctgtgat gcagaagaaa actatgggat gggaggcctc
     1981 caccgaaagg ctgtatccac gcgacggggt gctgaaagga gaaatccacc aggctctgaa
     2041 gctgaaagat gggggacatt acctggtgga gttcaagaca atctacatgg ccaagaaacc
     2101 tgtgcagctg ccaggctact attacgtgga cacaaaactg gatatcactt cacacaacga
     2161 ggactacact attgtggagc agtatgaacg gagcgagggg agacaccatc tgttcctggg
     2221 ccatgggact ggaagtaccg gctcagggtc tagtggaacc gcctcaagcg aggataacaa
     2281 tatggctgtg atcaaagagt tcatgaggtt taaggtgcgc atggagggca gcatgaatgg
     2341 gcacgaattt gagattgaag gagagggcga agggaggcct tacgagggca cacagactgc
     2401 caagctgaaa gtgaccaagg gaggaccact gcctttcgct tgggatatcc tgtctcctca
     2461 gtttatgtac ggaagtaagg cctatgtcaa gcatcccgct gacattcctg attacaagaa
     2521 actgtctttc ccagagggct ttaagtggga gagagtgatg aattttgaag atggaggcct
     2581 ggtgaccgtg acacaggact cctctctgca ggatggcact ctgatctaca aagtcaaaat
     2641 gcgcggcacc aattttccac ccgatgggcc cgtgatgcag aagaaaacaa tggggtggga
     2701 ggccagcact gaacggctgt atcctagaga cggagtgctg aagggcgaaa tccaccaggc
     2761 cctgaagctg aaagacggcg gccactacct ggtggagttc aaaaccatct acatggccaa
     2821 gaaaccagtg cagctgcccg gctattacta tgtggacacc aagctggata tcacatccca
     2881 caatgaagac tacaccattg tggaacagta tgagaggtct gaaggacgcc accatctgtt
     2941 tctgtacggc atggatgagc tgtataagtt ctgctacgag aatgaagtgt aatccggaat
     3001 caacctctgg attacaaaat ttgtgaaaga ttgactggta ttcttaacta tgttgctcct
     3061 tttacgctat gtggatacgc tgctttaatg cctttgtatc atgctattgc ttcccgtatg
     3121 gctttcattt tctcctcctt gtataaatcc tggttgctgt ctctttatga ggagttgtgg
     3181 cccgttgtca ggcaacgtgg cgtggtgtgc actgtgtttg ctgacgcaac ccccactggt
     3241 tggggcattg ccaccacctg tcagctcctt tccgggactt tcgctttccc cctccctatt
     3301 gccacggcgg aactcatcgc cgcctgcctt gcccgctgct ggacaggggc tcggctgttg
     3361 ggcactgaca attccgtggt gttgtcgggg aagctgacgt cctttccatg gctgctcgcc
     3421 tgtgttgcca cctggattct gcgcgggacg tccttctgct acgtcccttc ggccctcaat
     3481 ccagcggacc ttccttcccg cggcctgctg ccggctctgc ggcctcttcc gcgtcttcgc
     3541 cttcgccctc agacgagtcg gatctccctt tgggccgcct ccccgcctgg atcctgtgcc
     3601 ttctagttgc cagccatctg ttgtttgccc ctcccccgtg ccttccttga ccctggaagg
     3661 tgccactccc actgtccttt cctaataaaa tgaggaaatt gcatcgcatt gtctgagtag
     3721 gtgtcattct attctggggg gtggggtggg gcaggacagc aagggggagg attgggaaga
     3781 caatagcagg catgctgggg atgcggtggg ctctatggca ggaaccccta gtgatggagt
     3841 tggccactcc ctctctgcgc gctcgctcgc tcactgaggc cgggcgacca aaggtcgccc
     3901 gacgcccggg ctttgcccgg gcggcctcag tgagcgagcg agcgcgcagc tgcctgcagg
     3961 ggcgcctgat gcggtatttt ctccttacgc atctgtgcgg tatttcacac cgcatacgtc
     4021 aaagcaacca tagtacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac
     4081 gcgcagcgtg accgctacac ttgccagcgc cttagcgccc gctcctttcg ctttcttccc
     4141 ttcctttctc gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt
     4201 agggttccga tttagtgctt tacggcacct cgaccccaaa aaacttgatt tgggtgatgg
     4261 ttcacgtagt gggccatcgc cctgatagac ggtttttcgc cctttgacgt tggagtccac
     4321 gttctttaat agtggactct tgttccaaac tggaacaaca ctcaactcta tctcgggcta
     4381 ttcttttgat ttataaggga ttttgccgat ttcggtctat tggttaaaaa atgagctgat
     4441 ttaacaaaaa tttaacgcga attttaacaa aatattaacg tttacaattt tatggtgcac
     4501 tctcagtaca atctgctctg atgccgcata gttaagccag ccccgacacc cgccaacacc
     4561 cgctgacgcg ccctgacggg cttgtctgct cccggcatcc gcttacagac aagctgtgac
     4621 cgtctccggg agctgcatgt gtcagaggtt ttcaccgtca tcaccgaaac gcgcgagacg
     4681 aaagggcctc gtgatacgcc tatttttata ggttaatgtc atgataataa tggtttctta
     4741 gacgtcaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta
     4801 aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata
     4861 ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc gcccttattc ccttttttgc
     4921 ggcattttgc cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga
     4981 agatcagttg ggtgcacgag tgggttacat cgaactggat ctcaacagcg gtaagatcct
     5041 tgagagtttt cgccccgaag aacgttttcc aatgatgagc acttttaaag ttctgctatg
     5101 tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta
     5161 ttctcagaat gacttggttg agtactcacc agtcacagaa aagcatctta cggatggcat
     5221 gacagtaaga gaattatgca gtgctgccat aaccatgagt gataacactg cggccaactt
     5281 acttctgaca acgatcggag gaccgaagga gctaaccgct tttttgcaca acatggggga
     5341 tcatgtaact cgccttgatc gttgggaacc ggagctgaat gaagccatac caaacgacga
     5401 gcgtgacacc acgatgcctg tagcaatggc aacaacgttg cgcaaactat taactggcga
     5461 actacttact ctagcttccc ggcaacaatt aatagactgg atggaggcgg ataaagttgc
     5521 aggaccactt ctgcgctcgg cccttccggc tggctggttt attgctgata aatctggagc
     5581 cggtgagcgt gggtctcgcg gtatcattgc agcactgggg ccagatggta agccctcccg
     5641 tatcgtagtt atctacacga cggggagtca ggcaactatg gatgaacgaa atagacagat
     5701 cgctgagata ggtgcctcac tgattaagca ttggtaactg tcagaccaag tttactcata
     5761 tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct
     5821 ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga
     5881 ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg
     5941 cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc
     6001 aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct
     6061 agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc
     6121 tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt
     6181 ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg
     6241 cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct
     6301 atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag
     6361 ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag
     6421 tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg
     6481 gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg
     6541 gccttttgct cacatgt
//
