LOCUS       Exported                6152 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  AAV plasmid for expressing rsCaMPARI in neurons, nucleus excluded.
ACCESSION   .
VERSION     .
KEYWORDS    pAAV-hsyn_NES-His-rsCaMPARI-mRuby3
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6152)
  AUTHORS   Sha F, Abdelfattah AS, Patel R, Schreiter ER
  TITLE     Erasable labeling of neuronal activity using a reversible calcium 
            marker.
  JOURNAL   Elife. 2020 Sep 15;9. pii: 57249. doi: 10.7554/eLife.57249.
  PUBMED    32931424
REFERENCE   2  (bases 1 to 6152)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6152
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     primer_bind     131..150
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     repeat_region   309..438
                     /label=AAV2 ITR
     promoter        520..967
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     CDS             1064..1081
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             2324..3031
                     /codon_start=1
                     /product="monomeric red fluorescent protein derived from 
                     mRuby2, with improved brightness and photostability (Bajar 
                     et al., 2016)"
                     /label=mRuby3
                     /translation="VSKGEELIKENMRMKVVMEGSVNGHQFKCTGEGEGRPYEGVQTMR
                     IKVIEGGPLPFAFDILATSFMYGSRTFIKYPADIPDFFKQSFPEGFTWERVTRYEDGGV
                     VTVTQDTSLEDGELVYNVKVRGVNFPSNGPVMQKKTKGWEPNTEMMYPADGGLRGYTDI
                     ALKVDGGGHLHCNFVTTYRSKKTVGNIKMPGVHAVDHRLERIEESDNETYVVQREVAVA
                     KYSNLGGGMDELYK"
     misc_feature    3047..3635
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3100..3120)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3518..3529)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(3638..3654)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3639..3655)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     polyA_signal    3684..3805
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3695..3714)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3749..3768
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(4013..4030)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4184..4772)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4264..4283)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(4943..5803)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVMLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     5566..5585
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5804..5908)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(5934..237)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6071..6092)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
ORIGIN
        1 cactacgtga accatcaccc taatcaagtt ttttggggtc gaggtgccgt aaagcactaa
       61 atcggaaccc taaagggagc ccccgattta gagcttgacg gggaaagccg gcgaacgtgg
      121 cgagaaagga agggaagaaa gcgaaaggag cgggcgctag ggcgctggca agtgtagcgg
      181 tcacgctgcg cgtaaccacc acacccgccg cgcttaatgc gccgctacag ggcgcgtccc
      241 attcgccatt caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat
      301 tacgccagct gcgcgctcgc tcgctcactg aggccgcccg ggcaaagccc gggcgtcggg
      361 cgacctttgg tcgcccggcc tcagtgagcg agcgagcgcg cagagaggga gtggccaact
      421 ccatcactag gggttccttg tagttaatga ttaacccgcc atgctactta tctacgtagc
      481 catgctctag gaagatctct gcagagggcc ctgcgtatga gtgcaagtgg gttttaggac
      541 caggatgagg cggggtgggg gtgcctacct gacgaccgac cccgacccac tggacaagca
      601 cccaaccccc attccccaaa ttgcgcatcc cctatcagag agggggaggg gaaacaggat
      661 gcggcgaggc gcgtgcgcac tgccagcttc agcaccgcgg acagtgcctt cgcccccgcc
      721 tggcggcgcg cgccaccgcc gcctcagcac tgaaggcgcg ctgacgtcac tcgccggtcc
      781 cccgcaaact ccccttcccg gccaccttgg tcgcgtccgc gccgccgccg gcccagccgg
      841 accgcaccac gcgaggcgcg agataggggg gcacgggcgc gaccatctgc gctgcggcgc
      901 cggcgactca gcgctgcctc agtctgcggt gggcagcgga ggagtcgtgt cgtgcctgag
      961 agcgcagtcg aattcaagct gctagcaagg atccgccacc atgctgcaga acgagcttgc
     1021 tcttaagttg gctggacttg atattaacaa gactggaggt tctcatcatc accaccacca
     1081 tggatccatg gtgagtgcga ttaagccaga catgaagatc aaactgcgta tggaaggcaa
     1141 cgtaaacggg caccactttg tgatcgacgg agatggtaca ggcaagcctt atgagggaaa
     1201 acagaccatg gatcttgaag tcaaagaggg cggacctctg ccttttgcct ttgatatcct
     1261 gaccactgca ttcctgtacg gcaacagggt attcgtgaaa tatccagaca acatacaaga
     1321 ctattttaag cagtcgtttc ctaaggggta ttcgtgggaa cgaagcatga ctttcgaaga
     1381 cgggggcatt tgctatgcca gaagcgacat aacaatggaa ggggacactt tctataataa
     1441 agttcgattt tatggtacca actttcccgc caatggtcca gttatgcaga agaagacgct
     1501 gaaatgggag ccctccactg agaaaatgta tgtgcgtgat ggagtgctga cgggtgatgt
     1561 agagatggct ttgttgcttg aaggaaatgc ccattaccga gcgttgtcaa gccgccgcaa
     1621 gttcaacaaa accggccatg ccttgcgtgc aattggccgc ctgtccagcg gaggctctgg
     1681 cggttccggc ggttcaggtg gatcagacca gctgaccgag gagcaaattg cagaatttaa
     1741 ggaggcgttc tctttgtttg acaaagatgg cgacggtaca attacaacca aggaattggg
     1801 cacagtaatg cgctcccttg gacagaatcc cactgaagcc gaacttcaag acatgatcaa
     1861 tgaggttgat gctgacgggg atgggactat cgactttcct gagtttctta ccatgatggc
     1921 tcgcaaaatg aaggatacag acagtgagga ggaaattcgt gaggcctttc gtgtgtttga
     1981 taaggacggg aacggttata tctcagcggc ggagctgcgt cacgttatga ccaacttagg
     2041 cgaaaagctg actgatgaag aagtcgatga gatgatccgc gaggcggaca tcgatggtga
     2101 tggtcaagtc aattatgaag agtttgtggt gatgatgacc gcgaagtgcc tcttcagaac
     2161 tacttacaaa gctaaggaga agggtgtcaa gttaccaggc gtgcactatg tggaccacac
     2221 cattgagatt ttaagccatg acaaagatta caacaaggtt aagatctatg agtatgctgt
     2281 tgctcattct ggattgcctg acaatgccag acgaggaggt agcgtgtcta agggcgaaga
     2341 gctgatcaag gaaaatatgc gtatgaaggt ggtcatggaa ggttcggtca acggccacca
     2401 attcaaatgc acaggtgaag gagaaggcag accgtacgag ggagtgcaaa ccatgaggat
     2461 caaagtcatc gagggaggac ccctgccatt tgcctttgac attcttgcca cgtcgttcat
     2521 gtatggcagc cgtaccttta tcaagtaccc ggccgacatc cctgatttct ttaaacagtc
     2581 ctttcctgag ggttttactt gggaaagagt tacgagatac gaagatggtg gagtcgtcac
     2641 cgtcacgcag gacaccagcc ttgaggatgg cgagctcgtc tacaacgtca aggtcagagg
     2701 ggtaaacttt ccctccaatg gtcccgtgat gcagaagaag accaagggtt gggagcctaa
     2761 tacagagatg atgtatccag cagatggtgg tctgagagga tacactgaca tcgcactgaa
     2821 agttgatggt ggtggccatc tgcactgcaa cttcgtgaca acttacaggt caaaaaagac
     2881 cgtcgggaac atcaagatgc ccggtgtcca tgccgttgat caccgcctgg aaaggatcga
     2941 ggagagtgac aatgaaacct acgtagtgca aagagaagtg gcagttgcca aatacagcaa
     3001 ccttggtggt ggcatggacg agctgtacaa gtaaaagctt atcgataatc aacctctgga
     3061 ttacaaaatt tgtgaaagat tgactggtat tcttaactat gttgctcctt ttacgctatg
     3121 tggatacgct gctttaatgc ctttgtatca tgctattgct tcccgtatgg ctttcatttt
     3181 ctcctccttg tataaatcct ggttgctgtc tctttatgag gagttgtggc ccgttgtcag
     3241 gcaacgtggc gtggtgtgca ctgtgtttgc tgacgcaacc cccactggtt ggggcattgc
     3301 caccacctgt cagctccttt ccgggacttt cgctttcccc ctccctattg ccacggcgga
     3361 actcatcgcc gcctgccttg cccgctgctg gacaggggct cggctgttgg gcactgacaa
     3421 ttccgtggtg ttgtcgggga aatcatcgtc ctttccttgg ctgctcgcct gtgttgccac
     3481 ctggattctg cgcgggacgt ccttctgcta cgtcccttcg gccctcaatc cagcggacct
     3541 tccttcccgc ggcctgctgc cggctctgcg gcctcttccg cgtcttcgcc ttcgccctca
     3601 gacgagtcgg atctcccttt gggccgcctc cccgcatcga taccgtcgac ctcgacccgg
     3661 gcggccgctt cgagcagaca tgataagata cattgatgag tttggacaaa ccacaactag
     3721 aatgcagtga aaaaaatgct ttatttgtga aatttgtgat gctattgctt tatttgtaac
     3781 cattataagc tgcaataaac aagttaacaa caacaattgc attcatttta tgtttcaggt
     3841 tcagggggag atgtgggagg ttttttaaag caagtaaaac ctctacaaat gtggtaaaat
     3901 cgataaggat cttcctagag catggctacg tagataagta gcatggcggg ttaatcatta
     3961 actacaagga acccctagtg atggagttgg ccactccctc tctgcgcgct cgctcgctca
     4021 ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg
     4081 taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc
     4141 agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc
     4201 cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac
     4261 tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc
     4321 tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata
     4381 gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc
     4441 acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca
     4501 acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag
     4561 cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta
     4621 gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg
     4681 gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc
     4741 agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt
     4801 ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa
     4861 ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat
     4921 atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga
     4981 tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac
     5041 gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg
     5101 ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg
     5161 caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt
     5221 cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct
     5281 cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat
     5341 cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta
     5401 agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca
     5461 tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat
     5521 agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac
     5581 atagcataac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa
     5641 ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt
     5701 cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg
     5761 caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat
     5821 attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt
     5881 agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca cctaaattgt
     5941 aagcgttaat attttgttaa aattcgcgtt aaatttttgt taaatcagct cattttttaa
     6001 ccaataggcc gaaatcggca aaatccctta taaatcaaaa gaatagaccg agatagggtt
     6061 gagtgttgtt ccagtttgga acaagagtcc actattaaag aacgtggact ccaacgtcaa
     6121 agggcgaaaa accgtctatc agggcgatgg cc
//
