LOCUS       Exported                3736 bp ds-DNA     linear   SYN 18-MAY-2021
DEFINITION  Intersectional viral expression of GCaMP6M in cells expressing Cre 
            AND NOT Flp.
ACCESSION   .
VERSION     .
KEYWORDS    pAAV-Ef1a-Con/Foff 2.0-GCaMP6M
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3736)
  AUTHORS   Fenno LE, Ramakrishnan C, Kim YS, Evans KE, Lo M, Vesuna S, Inoue M,
            Cheung KYM, Yuen E, Pichamoorthy N, Hong ASO, Deisseroth K
  TITLE     Comprehensive Dual- and Triple-Feature Intersectional Single-Vector 
            Delivery of Diverse Functional Payloads to Cells of Behaving 
            Mammals.
  JOURNAL   Neuron. 2020 Jun 20. pii: S0896-6273(20)30432-3. doi: 
            10.1016/j.neuron.2020.06.003.
  PUBMED    32574559
REFERENCE   2  (bases 1 to 3736)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 18, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..3736
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     promoter        87..1264
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          318..1255
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     1212..1232
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     protein_bind    1271..1304
                     /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    1355..1388
                     /label=loxN
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GTATACCT). loxN sites are compatible with each 
                     other, but incompatible with loxP or lox2272 sites."
     primer_bind     complement(1481..1502)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             complement(1715..1771)
                     /codon_start=1
                     /product="calmodulin-binding peptide from avian smooth 
                     muscle myosin light chain kinase"
                     /label=calmodulin-binding peptide
                     /translation="RRKWNKTGHAVRAIGRLSS"
     regulatory      1783..1792
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     protein_bind    complement(1799..1832)
                     /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(1883..1916)
                     /label=loxN
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GTATACCT). loxN sites are compatible with each 
                     other, but incompatible with loxP or lox2272 sites."
     protein_bind    1923..1956
                     /label=FRT (minimal)
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="supports FLP-mediated excision but not integration 
                     (Turan and Bode, 2011)"
     primer_bind     complement(2150..2171)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2411..2430)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     protein_bind    complement(2996..3029)
                     /label=FRT (minimal)
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="supports FLP-mediated excision but not integration 
                     (Turan and Bode, 2011)"
     misc_feature    3138..3726
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3191..3211)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3609..3620)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
ORIGIN
        1 aagctttgca aagatggata aagttttaaa cagagaggaa tctttgcagc taatggacct
       61 tctaggtctt gaaaggagtg ggaattggct ccggtgcccg tcagtgggca gagcgcacat
      121 cgcccacagt ccccgagaag ttggggggag gggtcggcaa ttgaaccggt gcctagagaa
      181 ggtggcgcgg ggtaaactgg gaaagtgatg tcgtgtactg gctccgcctt tttcccgagg
      241 gtgggggaga accgtatata agtgcagtag tcgccgtgaa cgttcttttt cgcaacgggt
      301 ttgccgccag aacacaggta agtgccgtgt gtggttcccg cgggcctggc ctctttacgg
      361 gttatggccc ttgcgtgcct tgaattactt ccactggctg cagtacgtga ttcttgatcc
      421 cgagcttcgg gttggaagtg ggtgggagag ttcgaggcct tgcgcttaag gagccccttc
      481 gcctcgtgct tgagttgagg cctggcctgg gcgctggggc cgccgcgtgc gaatctggtg
      541 gcaccttcgc gcctgtctcg ctgctttcga taagtctcta gccatttaaa atttttgatg
      601 acctgctgcg acgctttttt tctggcaaga tagtcttgta aatgcgggcc aagatctgca
      661 cactggtatt tcggtttttg gggccgcggg cggcgacggg gcccgtgcgt cccagcgcac
      721 atgttcggcg aggcggggcc tgcgagcgcg gccaccgaga atcggacggg ggtagtctca
      781 agctggccgg cctgctctgg tgcctggcct cgcgccgccg tgtatcgccc cgccctgggc
      841 ggcaaggctg gcccggtcgg caccagttgc gtgagcggaa agatggccgc ttcccggccc
      901 tgctgcaggg agctcaaaat ggaggacgcg gcgctcggga gagcgggcgg gtgagtcacc
      961 cacacaaagg aaaagggcct ttccgtcctc agccgtcgct tcatgtgact ccacggagta
     1021 ccgggcgccg tccaggcacc tcgattagtt ctcgagcttt tggagtacgt cgtctttagg
     1081 ttggggggag gggttttatg cgatggagtt tccccacact gagtgggtgg agactgaagt
     1141 taggccagct tggcacttga tgtaattctc cttggaattt gccctttttg agtttggatc
     1201 ttggttcatt ctcaagcctc agacagtggt tcaaagtttt tttcttccat ttcaggtgtc
     1261 gtgaggatcc ataacttcgt ataggatact ttatacgaag ttatgtagct ggagcaattt
     1321 gaaaagattg ttgtattacc tctagctgcg cttaataact tcgtatagta taccttatac
     1381 gaagttatgt cgaccctcct gtactcacct tgctcaccat gctccctccg gtaccgccct
     1441 tgtacagctc gtccatgccg agagtgatcc cggcggcggt cacgaactcc agcaggacca
     1501 tgtgatcgcg cttctcgttg gggtctttcg aaagtttgga ctgcacgctc aggtagtggt
     1561 tgtcgggcag cagcacgggg ccgtcgccga tgggggtgtt ctgctggtag tggtaggcga
     1621 gctgcacgcc gccgtcctcg atgttgtggc ggatcttgaa gttcgccttg atgccgttct
     1681 tctgcttgtc ggccttgata tagacgttct cgagtgagct cagccgacct atagctctga
     1741 ctgcgtgacc tgtcttattc cacttacgac gtgatgagtc caccatggtg gcactagtat
     1801 aacttcgtat aaagtatcct atacgaagtt atttgcctta acccagaaat tatcactgtt
     1861 attctttaga taggtgcaaa gaataacttc gtataaggta tactatacga agttatggta
     1921 ccgaagttcc tattctctag aaagtatagg aacttcgcag aatggtagct ggattgtagc
     1981 tgctattagc aatatgaaac ctcttagaag ttcctattct tcaaaaggta taggaacttc
     2041 ggcgcgcctg gagagtggcc agcccttctc atgttcagag aacatggtta actggttaag
     2101 tcatgtcgtc ccacagggcg aggagctgtt caccggggtg gtgcccatcc tggtcgagct
     2161 ggacggcgac gtaaacggcc acaagttcag cgtgtccggc gagggtgagg gcgatgccac
     2221 ctacggcaag ctgaccctga agttcatctg caccaccggc aagctgcccg tgccctggcc
     2281 caccctcgtg accaccctga cctacggcgt gcagtgcttc agccgctacc ccgaccacat
     2341 gaagcagcac gacttcttca agtccgccat gcccgaaggc tacatccagg agcgcaccat
     2401 cttcttcaag gacgacggca actacaagac ccgcgccgag gtgaagttcg agggcgacac
     2461 cctggtgaac cgcatcgagc tgaagggcat cgacttcaag gaggacggca acatcctggg
     2521 gcacaagctg gagtacaacc tgccggacca actgactgaa gagcagatcg cagaatttaa
     2581 agaggctttc tccctatttg acaaggacgg ggatgggaca ataacaacca aggagctggg
     2641 gacggtgatg cggtctctgg ggcagaaccc cacagaagca gagctgcagg acatgatcaa
     2701 tgaagtagat gccgacggtg acggcacaat cgacttccct gagttcctga caatgatggc
     2761 aagaaaaggg agctacaggg acacggaaga agaaattaga gaagcgttcg gtgtgtttga
     2821 taaggatggc aatggctaca tcagtgcagc agagcttcgc cacgtgatga caaaccttgg
     2881 agagaagtta acagatgaag aggttgatga aatgatcagg gaagcagaca tcgatgggga
     2941 tggtcaggta aactacgaag agtttgtaca aatgatgaca gcgaagtgag ctagcgaagt
     3001 tcctatactt tctagagaat aggaacttct aagcagacaa tcttgtaata catcagcagc
     3061 tagtagattg ctccagctag aagttcctat accttttgaa gaataggaac ttcgaattcg
     3121 atatcaagct tatcgataat caacctctgg attacaaaat ttgtgaaaga ttgactggta
     3181 ttcttaacta tgttgctcct tttacgctat gtggatacgc tgctttaatg cctttgtatc
     3241 atgctattgc ttcccgtatg gctttcattt tctcctcctt gtataaatcc tggttgctgt
     3301 ctctttatga ggagttgtgg cccgttgtca ggcaacgtgg cgtggtgtgc actgtgtttg
     3361 ctgacgcaac ccccactggt tggggcattg ccaccacctg tcagctcctt tccgggactt
     3421 tcgctttccc cctccctatt gccacggcgg aactcatcgc cgcctgcctt gcccgctgct
     3481 ggacaggggc tcggctgttg ggcactgaca attccgtggt gttgtcgggg aaatcatcgt
     3541 cctttccttg gctgctcgcc tatgttgcca cctggattct gcgcgggacg tccttctgct
     3601 acgtcccttc ggccctcaat ccagcggacc ttccttcccg cggcctgctg ccggctctgc
     3661 ggcctcttcc gcgtcttcgc cttcgccctc agacgagtcg gatctccctt tgggccgcct
     3721 ccccgcatcg ataccg
//
