LOCUS       Exported                6740 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  AAV-mediated expression of Jaws under the CaMKII promoter.
ACCESSION   .
VERSION     .
KEYWORDS    pAAV-CaMKII-Jaws-KGC-GFP-ER2
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6740)
  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 6740)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6740
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     repeat_region   62..191
                     /label=AAV2 ITR
     promoter        264..1552
                     /label=CaMKII promoter
                     /note="promoter for the mouse calcium/calmodulin-dependent 
                     kinase II alpha subunit"
     CDS             2483..3196
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(2525..2546)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2786..2805)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3133..3154
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    3245..3833
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3298..3318)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3716..3727)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3876..3997
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3887..3906)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3941..3960
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     repeat_region   4159..4288
                     /label=AAV2 ITR
     primer_bind     complement(4354..4371)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4525..5113)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4605..4624)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(5284..6144)
                     /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     5907..5926
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6145..6249)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(6275..6730)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6412..6433)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6624..6643
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 attcaggctg cgcaactgtt gggaagggcg atcggtgcgg gcctcttcgc tattacgcca
       61 gctgcgcgct cgctcgctca ctgaggccgc ccgggcaaag cccgggcgtc gggcgacctt
      121 tggtcgcccg gcctcagtga gcgagcgagc gcgcagagag ggagtggcca actccatcac
      181 taggggttcc ttgtagttaa tgattaaccc gccatgctac ttatctacgt agccatgctc
      241 taggaagatc gtaggttaat taacattatg gccttaggtc acttcatctc catggggttc
      301 ttcttctgat tttctagaaa atgagatggg ggtgcagaga gcttcctcag tgacctgccc
      361 agggtcacat cagaaatgtc agagctagaa cttgaactca gattactaat cttaaattcc
      421 atgccttggg ggcatgcaag tacgatatac agaaggagtg aactcattag ggcagatgac
      481 caatgagttt aggaaagaag agtccagggc agggtacatc tacaccaccc gcccagccct
      541 gggtgagtcc agccacgttc acctcattat agttgcctct ctccagtcct accttgacgg
      601 gaagcacaag cagaaactgg gacaggagcc ccaggagacc aaatcttcat ggtccctctg
      661 ggaggatggg tggggagagc tgtggcagag gcctcaggag gggccctgct gctcagtggt
      721 gacagatagg ggtgagaaag cagacagagt cattccgtca gcattctggg tctgtttggt
      781 acttcttctc acgctaaggt ggcggtgtga tatgcacaat ggctaaaaag cagggagagc
      841 tggaaagaaa caaggacaga gacagaggcc aagtcaacca gaccaattcc cagaggaagc
      901 aaagaaacca ttacagagac tacaaggggg aagggaagga gagatgaatt agcttcccct
      961 gtaaacctta gaacccagct gttgccaggg caacggggca atacctgtct cttcagagga
     1021 gatgaagttg ccagggtaac tacatcctgt ctttctcaag gaccatccca gaatgtggca
     1081 cccactagcc gttaccatag caactgcctc tttgccccac ttaatcccat cccgtctgtt
     1141 aaaagggccc tatagttgga ggtgggggag gtaggaagag cgatgatcac ttgtggacta
     1201 agtttgttcg catccccttc tccaaccccc tcagtacatc accctggggg aacagggtcc
     1261 acttgctcct gggcccacac agtcctgcag tattgtgtat ataaggccag ggcaaagagg
     1321 agcaggtttt aaagtgaaag gcaggcaggt gttggggagg cagttaccgg ggcaacggga
     1381 acagggcgtt tcggaggtgg ttgccatggg gacctggatg ctgacgaagg ctcgcgaggc
     1441 tgtgagcagc cacagtgccc tgctcagaag ccccaagctc gtcagtcaag ccggttctcc
     1501 gtttgcactc aggagcacgg gcaggcgagt ggcccctagt tctgggggca gcgggggatc
     1561 cgccaccatg accgccgtga gcaccacagc cactaccgtg ctgcaggcca cacagagcga
     1621 cgtgctgcag gagatccagt ccaacttcct gctgaatagc tccatctggg tgaacattgc
     1681 tctggccgga gtggtcatcc tgctgtttgt ggccatgggg agggatctgg aatcccctag
     1741 agctaagctg atctgggtgg ccacaatgct ggtgccactg gtgtctattt ctagttacgc
     1801 tggactggcc agtgggctga ctgtgggctt cctgcagatg ccacctggac acgctctggc
     1861 cggacaggag gtgctgagcc catggggccg gtatctgaca tggactttct ccactcccat
     1921 gatcctgctg gctctgggac tgctggccga caccgatatt gccagcctgt tcaccgccat
     1981 cacaatggac attggcatgt gcgtgacagg actggccgct gccctgatca ctagctccca
     2041 tctgctgcgc tgggtgttct acggaatttc ttgtgctttc tttgtggccg tgctgtatgt
     2101 gctgctggtg cagtggccag ctgatgctga ggctgctggg accagtgaaa tctttggcac
     2161 tctgaggatt ctgaccgtgg tgctgtggct ggggtaccct atcctgttcg ctctgggctc
     2221 tgagggagtg gccctgctga gtgtgggagt gaccagctgg ggatactccg gactggacat
     2281 cctggctaaa tacgtgttcg cctttctgct gctgagatgg gtggctgcca atgaaggcac
     2341 agtgtctggg agtggaatgg gaatcgggtc cggaggagct gctccagccg acgatcgacc
     2401 ggtagtaaaa tccagaatta cttctgaagg ggagtatatc cctctggatc aaatagacat
     2461 caatgttgca cctgcagggg cagtgagcaa gggcgaggag ctgttcaccg gggtggtgcc
     2521 catcctggtc gagctggacg gcgacgtaaa cggccacaag ttcagcgtgt ccggcgaggg
     2581 cgagggcgat gccacctacg gcaagctgac cctgaagttc atttgcacca ccggcaagct
     2641 gcccgtgccc tggcccaccc tcgtgaccac cctgacctac ggcgtgcagt gcttcagccg
     2701 ctaccccgac cacatgaagc agcacgactt cttcaagtcc gccatgcccg aaggctacgt
     2761 ccaggagcgc accatcttct tcaaggacga cggcaactac aagacccgcg ccgaggtgaa
     2821 gttcgagggc gacaccctgg tgaaccgcat cgagctgaag ggcatcgact tcaaggagga
     2881 cggcaacatc ctggggcaca agctggagta caactacaac agccacaacg tctatatcat
     2941 ggccgacaag cagaagaacg gcatcaaggt gaacttcaag atccgccaca acatcgagga
     3001 cggcagcgtg cagctcgccg accactacca gcagaacacc cccatcggcg acggccccgt
     3061 gctgctgccc gacaaccact acctgagcac ccagtccgcc ctgagcaaag accccaacga
     3121 gaagcgcgat cacatggtcc tgctggagtt cgtgaccgcc gccgggatca ctctcggcat
     3181 ggacgagctg tacaagttct gctacgagaa tgaagtgtaa gaattcgata tcaagcttat
     3241 cgataatcaa cctctggatt acaaaatttg tgaaagattg actggtattc ttaactatgt
     3301 tgctcctttt acgctatgtg gatacgctgc tttaatgcct ttgtatcatg ctattgcttc
     3361 ccgtatggct ttcattttct cctccttgta taaatcctgg ttgctgtctc tttatgagga
     3421 gttgtggccc gttgtcaggc aacgtggcgt ggtgtgcact gtgtttgctg acgcaacccc
     3481 cactggttgg ggcattgcca ccacctgtca gctcctttcc gggactttcg ctttccccct
     3541 ccctattgcc acggcggaac tcatcgccgc ctgccttgcc cgctgctgga caggggctcg
     3601 gctgttgggc actgacaatt ccgtggtgtt gtcggggaaa tcatcgtcct ttccttggct
     3661 gctcgcctgt gttgccacct ggattctgcg cgggacgtcc ttctgctacg tcccttcggc
     3721 cctcaatcca gcggaccttc cttcccgcgg cctgctgccg gctctgcggc ctcttccgcg
     3781 tcttcgcctt cgccctcaga cgagtcggat ctccctttgg gccgcctccc cgcatcgata
     3841 ccgtcgaccc gggcggccgc ttcgagcaga catgataaga tacattgatg agtttggaca
     3901 aaccacaact agaatgcagt gaaaaaaatg ctttatttgt gaaatttgtg atgctattgc
     3961 tttatttgta accattataa gctgcaataa acaagttaac aacaacaatt gcattcattt
     4021 tatgtttcag gttcaggggg agatgtggga ggttttttaa agcaagtaaa acctctacaa
     4081 atgtggtaaa atcgataagg atcttcctag agcatggcta cgtagataag tagcatggcg
     4141 ggttaatcat taactacaag gaacccctag tgatggagtt ggccactccc tctctgcgcg
     4201 ctcgctcgct cactgaggcc gggcgaccaa aggtcgcccg acgcccgggc tttgcccggg
     4261 cggcctcagt gagcgagcga gcgcgcagct gcattaatga atcggccaac gcgcggggag
     4321 aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt
     4381 cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga
     4441 atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg
     4501 taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa
     4561 aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt
     4621 tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct
     4681 gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct
     4741 cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc
     4801 cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt
     4861 atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc
     4921 tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat
     4981 ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa
     5041 acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa
     5101 aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga
     5161 aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct
     5221 tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga
     5281 cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc
     5341 catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg
     5401 ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat
     5461 aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat
     5521 ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg
     5581 caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc
     5641 attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa
     5701 agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc
     5761 actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt
     5821 ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag
     5881 ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt
     5941 gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag
     6001 atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac
     6061 cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc
     6121 gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca
     6181 gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg
     6241 ggttccgcgc acatttcccc gaaaagtgcc acctaaattg taagcgttaa tattttgtta
     6301 aaattcgcgt taaatttttg ttaaatcagc tcatttttta accaataggc cgaaatcggc
     6361 aaaatccctt ataaatcaaa agaatagacc gagatagggt tgagtgttgt tccagtttgg
     6421 aacaagagtc cactattaaa gaacgtggac tccaacgtca aagggcgaaa aaccgtctat
     6481 cagggcgatg gcccactacg tgaaccatca ccctaatcaa gttttttggg gtcgaggtgc
     6541 cgtaaagcac taaatcggaa ccctaaaggg agcccccgat ttagagcttg acggggaaag
     6601 ccggcgaacg tggcgagaaa ggaagggaag aaagcgaaag gagcgggcgc tagggcgctg
     6661 gcaagtgtag cggtcacgct gcgcgtaacc accacacccg ccgcgcttaa tgcgccgcta
     6721 cagggcgcgt cccattcgcc
//
