LOCUS       Exported                5972 bp ds-DNA     circular SYN 13-MAY-2021
DEFINITION  AAV expression of Cre recombinase-activated jRCaMP1a, a red 
            fluorescent calcium sensor protein, from the Synapsin promoter.
ACCESSION   .
VERSION     .
KEYWORDS    pAAV.Syn.Flex.NES-jRCaMP1a.WPRE.SV40
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5972)
  AUTHORS   Dana H, Mohar B, Sun Y, Narayan S, Gordus A, Hasseman JP, Tsegaye G,
            Holt GT, Hu A, Walpita D, Patel R, Macklin JJ, Bargmann CI, Ahrens 
            MB, Schreiter ER, Jayaraman V, Looger LL, Svoboda K, Kim DS
  TITLE     Sensitive red protein calcium indicators for imaging neural 
            activity.
  JOURNAL   Elife. 2016 Mar 24;5. pii: e12727. doi: 10.7554/eLife.12727.
  PUBMED    27011354
REFERENCE   2  (bases 1 to 5972)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 13, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..5972
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     promoter        211..658
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     protein_bind    complement(688..721)
                     /label=lox2272
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GGATACTT). lox2272 sites are compatible with each
                     other, but incompatible with loxP or loxN sites (Lee and 
                     Saito, 1988)."
     protein_bind    complement(784..817)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GCATACAT)."
     CDS             complement(1983..2039)
                     /codon_start=1
                     /product="calmodulin-binding peptide from avian smooth 
                     muscle myosin light chain kinase"
                     /label=calmodulin-binding peptide
                     /translation="RRKWNKTGHAVRAIGRLSS"
     CDS             complement(2067..2090)
                     /codon_start=1
                     /product="Xpress(TM) epitope tag, including an enterokinase
                     recognition and cleavage site"
                     /label=Xpress(TM) tag
                     /translation="DLYDDDDK"
     CDS             complement(2094..2126)
                     /codon_start=1
                     /product="leader peptide from bacteriophage T7 gene 10"
                     /label=T7 tag (gene 10 leader)
                     /note="promotes efficient translation in E. coli"
                     /translation="MASMTGGQQMG"
     CDS             complement(2130..2147)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     protein_bind    2230..2263
                     /label=lox2272
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GGATACTT). lox2272 sites are compatible with each
                     other, but incompatible with loxP or loxN sites (Lee and 
                     Saito, 1988)."
     protein_bind    2334..2367
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GCATACAT)."
     misc_feature    2409..2997
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2462..2482)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2880..2891)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(3000..3016)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3001..3017)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     polyA_signal    3046..3167
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3057..3076)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3111..3130
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     repeat_region   3329..3458
                     /label=AAV2 ITR
     rep_origin      3530..3985
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3617..3636)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3827..3848
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        4011..4115
                     /gene="bla"
                     /label=AmpR promoter
     CDS             4116..4976
                     /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(4334..4353)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5147..5735
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5636..5655
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5889..5906
                     /label=L4440
                     /note="L4440 vector, forward primer"
     repeat_region   5972..129
                     /label=AAV2 ITR
ORIGIN
        1 tgcgcgctcg ctcgctcact gaggccgccc gggcaaagcc cgggcgtcgg gcgacctttg
       61 gtcgcccggc ctcagtgagc gagcgagcgc gcagagaggg agtggccaac tccatcacta
      121 ggggttcctt gtagttaatg attaacccgc catgctactt atctacgtag ccatgctcta
      181 ggaagatctc tgcagagggc cctgcgtatg agtgcaagtg ggttttagga ccaggatgag
      241 gcggggtggg ggtgcctacc tgacgaccga ccccgaccca ctggacaagc acccaacccc
      301 cattccccaa attgcgcatc ccctatcaga gagggggagg ggaaacagga tgcggcgagg
      361 cgcgtgcgca ctgccagctt cagcaccgcg gacagtgcct tcgcccccgc ctggcggcgc
      421 gcgccaccgc cgcctcagca ctgaaggcgc gctgacgtca ctcgccggtc ccccgcaaac
      481 tccccttccc ggccaccttg gtcgcgtccg cgccgccgcc ggcccagccg gaccgcacca
      541 cgcgaggcgc gagatagggg ggcacgggcg cgaccatctg cgctgcggcg ccggcgactc
      601 agcgctgcct cagtctgcgg tgggcagcgg aggagtcgtg tcgtgcctga gagcgcagtc
      661 gaattcaagc tgctagcaag gatccccata acttcgtata aagtatccta tacgaagtta
      721 tatcaaaata ggaagaccaa tgcttcacca tcgacccgaa ttgccaagca tcaccatcga
      781 cccataactt cgtataatgt atgctatacg aagttatact agagctttca cttcgctgtc
      841 atcatttgta caaactcttc gtagtttacc tgaccatccc catcgctgtc tgcttccctg
      901 atcatttcat caacctcttc atctgttaac ttctctccaa ggtttgtcat cacgtggcga
      961 agctctgctg cactgatgta gccattgcca tccttatcaa acacgccgaa cgcttctcta
     1021 atttcttctt cactgtctgt gtatttcatt tttcttgcca tcataatcag gaactcaggg
     1081 aagtcgattg tgccgtcacc gtcggcatct acttcattga tcatgtcctg cagctctgct
     1141 tctgtggggt tctgccccag agaccgcatc accgtcccca gctccttggt tgttattgtc
     1201 ccatccccgt ccttgtcaaa tagggagaaa gcctctttaa attctgcgat ctgctcttca
     1261 gtggtttggg agtccgtagg ctcccaacct ttggttttct tttgcattac agcaccattg
     1321 ctcgggaagt tcacaccgcg aacctgcacg tgatacacca ggcagccatc ctccagagac
     1381 gtgtcttgca taacggtaat tacgccgccg tcttcgtaac gggtcacacg ttcccaggta
     1441 aaaccttccg ggaaggactg cttaaagaaa tccgggatgc ctttcggata tttgatgaag
     1501 gtacgggagc catacataaa ggacgtcgcc aggatgtcga acgcaaacgg cagcggacca
     1561 ccctcgatca ctttgatacg catagtctgg gtgcccatat acgggttgcc ttcgccttca
     1621 ccagtgcatt tgaactggtg gccgtttacg gagccttcca gaaccacttt catacgcatg
     1681 ttctccttga tcagggagtt catgctccct ccggtaccgc cgccaccacc caggcccaca
     1741 aacttggcaa ctgcgtgttc acgctgtacg acaaacattt cgttatcgct ctcctccagg
     1801 cgctccagac ggtggctgac gctatgaatg gcaggcatct taatgttgcc gacagtcttt
     1861 ttggagcggt aggtggtcac gaaagaacag gacaggtgac cgccgccatc aactttcagc
     1921 gccatgtgag tgtaaccacg cagaccacca tccgctgggt acatcatttc gcagttgatc
     1981 gctgagctca gccgacctat agctctgact gcgtgacctg tcttattcca cttacgacgt
     2041 gatgagtcga ccattgttgc gagatcctta tcgtcatcgt cgtacagatc ccgacccatt
     2101 tgctgtccac cagtcatgct agccatacca tgatgatgat gatgatgaga acctcctcca
     2161 gtcttgttaa tatcaagtcc agccaactta agagcaagct cgttctgcag catggtggcg
     2221 gatcctagta taacttcgta taggatactt tatacgaagt tatcattggg attcttccta
     2281 ttttgatcca agcatcacca tcgaccctct agtccagatc tcaccatcga cccataactt
     2341 cgtatagcat acattatacg aagttatgtc cctcgaagag gttcgaattc gatatcaagc
     2401 ttatcgataa tcaacctctg gattacaaaa tttgtgaaag attgactggt attcttaact
     2461 atgttgctcc ttttacgcta tgtggatacg ctgctttaat gcctttgtat catgctattg
     2521 cttcccgtat ggctttcatt ttctcctcct tgtataaatc ctggttgctg tctctttatg
     2581 aggagttgtg gcccgttgtc aggcaacgtg gcgtggtgtg cactgtgttt gctgacgcaa
     2641 cccccactgg ttggggcatt gccaccacct gtcagctcct ttccgggact ttcgctttcc
     2701 ccctccctat tgccacggcg gaactcatcg ccgcctgcct tgcccgctgc tggacagggg
     2761 ctcggctgtt gggcactgac aattccgtgg tgttgtcggg gaaatcatcg tcctttcctt
     2821 ggctgctcgc ctgtgttgcc acctggattc tgcgcgggac gtccttctgc tacgtccctt
     2881 cggccctcaa tccagcggac cttccttccc gcggcctgct gccggctctg cggcctcttc
     2941 cgcgtcttcg ccttcgccct cagacgagtc ggatctccct ttgggccgcc tccccgcatc
     3001 gataccgtcg acctcgaccc gggcggccgc ttcgagcaga catgataaga tacattgatg
     3061 agtttggaca aaccacaact agaatgcagt gaaaaaaatg ctttatttgt gaaatttgtg
     3121 atgctattgc tttatttgta accattataa gctgcaataa acaagttaac aacaacaatt
     3181 gcattcattt tatgtttcag gttcaggggg agatgtggga ggttttttaa agcaagtaaa
     3241 acctctacaa atgtggtaaa atcgataagg atcttcctag agcatggcta cgtagataag
     3301 tagcatggcg ggttaatcat taactacaag gaacccctag tgatggagtt ggccactccc
     3361 tctctgcgcg ctcgctcgct cactgaggcc gggcgaccaa aggtcgcccg acgcccgggc
     3421 tttgcccggg cggcctcagt gagcgagcga gcgcgcagct ggcgtaatag cgaagaggcc
     3481 cgcaccgatc gcccttccca acagttgcgc agcctgaatg gcgaatggga cgcgccctgt
     3541 agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc
     3601 agcgccctag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc
     3661 tttccccgtc aagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg
     3721 cacctcgacc ccaaaaaact tgattagggt gatggttcac gtagtgggcc atcgccctga
     3781 tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc
     3841 caaactggaa caacactcaa ccctatctcg gtctattctt ttgatttata agggattttg
     3901 ccgatttcgg cctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaatttt
     3961 aacaaaatat taacgcttac aatttaggtg gcacttttcg gggaaatgtg cgcggaaccc
     4021 ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct
     4081 gataaatgct tcaataatat tgaaaaagga agagtatgag tattcaacat ttccgtgtcg
     4141 cccttattcc cttttttgcg gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg
     4201 tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt gggttacatc gaactggatc
     4261 tcaacagcgg taagatcctt gagagttttc gccccgaaga acgttttcca atgatgagca
     4321 cttttaaagt tctgctatgt ggcgcggtat tatcccgtat tgacgccggg caagagcaac
     4381 tcggtcgccg catacactat tctcagaatg acttggttga gtactcacca gtcacagaaa
     4441 agcatcttac ggatggcatg acagtaagag aattatgcag tgctgccata accatgagtg
     4501 ataacactgc ggccaactta cttctgacaa cgatcggagg accgaaggag ctaaccgctt
     4561 ttttgcacaa catgggggat catgtaactc gccttgatcg ttgggaaccg gagctgaatg
     4621 aagccatacc aaacgacgag cgtgacacca cgatgcctgt agcaatggca acaacgttgc
     4681 gcaaactatt aactggcgaa ctacttactc tagcttcccg gcaacaatta atagactgga
     4741 tggaggcgga taaagttgca ggaccacttc tgcgctcggc ccttccggct ggctggttta
     4801 ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc
     4861 cagatggtaa gccctcccgt atcgtagtta tctacacgac ggggagtcag gcaactatgg
     4921 atgaacgaaa tagacagatc gctgagatag gtgcctcact gattaagcat tggtaactgt
     4981 cagaccaagt ttactcatat atactttaga ttgatttaaa acttcatttt taatttaaaa
     5041 ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt
     5101 cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt
     5161 ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt
     5221 tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga
     5281 taccaaatac tgttcttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag
     5341 caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata
     5401 agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg
     5461 gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga
     5521 gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca
     5581 ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa
     5641 acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt
     5701 tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac
     5761 ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta tcccctgatt
     5821 ctgtggataa ccgtattacc gcctttgagt gagctgatac cgctcgccgc agccgaacga
     5881 ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg cccaatacgc aaaccgcctc
     5941 tccccgcgcg ttggccgatt cattaatgca gc
//
