LOCUS       Exported                3585 bp ds-DNA     linear   SYN 14-JUN-2021
DEFINITION  Intersectional viral expression of bREACHes-EYFP in cells expressing
            Cre AND NOT Flp.
ACCESSION   .
VERSION     .
KEYWORDS    pAAV-nEF-Con/Foff 2.0-bREACHes-EYFP
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3585)
  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 3585)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Jun 14, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..3585
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     promoter        33..244
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     LTR             257..525
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from human 
                     T-cell leukemia virus (HTLV) type 1"
     protein_bind    553..586
                     /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)"
     regulatory      679..688
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     protein_bind    1165..1198
                     /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    1249..1282
                     /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)."
     primer_bind     1395..1416
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     protein_bind    complement(2086..2119)
                     /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    complement(2170..2203)
                     /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)."
     primer_bind     complement(2425..2444)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     CDS             2584..2835
                     /codon_start=1
                     /product="C-terminal fragment of mVenus for use in 
                     bimolecular fluorescence complementation (BiFC) (Kodama and
                     Hu, 2010)"
                     /label=VC155
                     /translation="DKQKNGIKVNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNH
                     YLSYQSALSKDPNEKRDHMVLLEFVTAAGITLGMDELYK"
     primer_bind     2772..2793
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     protein_bind    complement(2845..2878)
                     /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    2987..3575
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3040..3060)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3458..3469)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
ORIGIN
        1 aaggatctgc gatcgctccg gtgcccgtca gtgggcagag cgcacatcgc ccacagtccc
       61 cgagaagttg gggggagggg tcggcaattg aacgggtgcc tagagaaggt ggcgcggggt
      121 aaactgggaa agtgatgtcg tgtactggct ccgccttttt cccgagggtg ggggagaacc
      181 gtatataagt gcagtagtcg ccgtgaacgt tctttttcgc aacgggtttg ccgccagaac
      241 acagctgaag cttcgagggg ctcgcatctc tccttcacgc gcccgccgcc ctacctgagg
      301 ccgccatcca cgccggttga gtcgcgttct gccgcctccc gcctgtggtg cctcctgaac
      361 tgcgtccgcc gtctaggtaa gtttaaagct caggtcgaga ccgggccttt gtccggcgct
      421 cccttggagc ctacctagac tcagccggct ctccacgctt tgcctgaccc tgcttgctca
      481 actctacgtc tttgtttcgt tttctgttct gcgccgttac agatccaagc tgtgaccggc
      541 gcctacggat ccgaagttcc tattctctag aaagtatagg aacttcgcag agtagtagct
      601 ggattgtagc tgctattagc aatctaaaac ctcttagaag ttcctattct tcaaaaggta
      661 taggaacttc ggcgcgccgc caccatggac tatggcggcg ctttgtctgc cgtcggactg
      721 ttccagacca gctacaccct ggagaacaac ggcagcgtga tctgcatccc caacaacggc
      781 cagtgcttct gcctggcctg gctgaagagt aacggcacca acgccgagaa gctggccgcc
      841 aacatcctgc agtgggtggt gtttgcgctg agcgtggcgt gcctgggctg gtatgcgtat
      901 caggcgtggc gcgcgacctg cggctgggaa gaagtgtatg tggcgctgat tgaaatgatg
      961 aaaagcatta ttgaagcgtt tcatgaattt gatagcccgg cgaccctgtg gctgagcagc
     1021 ggcaacggcg tggtgtggat gcgctatggc tcatggctgc tgacctgccc cgtgattctg
     1081 attcatctga gcaacctgac cggcctgaaa gatgattata gcaaacgcac catgggcctg
     1141 ctggtaagtg tcggggttga gctcataact tcgtatagta taccttatac gaagttattc
     1201 tttgcaccat tctaaagaat aacagtgata atttctgggt taaggcaaat aacttcgtat
     1261 aggatacttt atacgaagtt atggtacccc tcctgtactc accttgccgg tggtgcagat
     1321 gaacttcagg gtcagcttgc cgtaggtggc atcgccctcg ccctcgccgg acacgctgaa
     1381 cttgtggccg tttacgtcgc cgtccagctc gaccaggatg ggcaccaccc cggtgaacag
     1441 ctcctcgccc ttgctcacca cgttgatgtc gatctggtcc agggggatgt actcgccctc
     1501 gctggtgatc ctgctcttgg cggccgcatc ttcttcttcc gccaccaggg tttccacttc
     1561 catttcctgg cccgcaatgg taattttctg ttttttgcga atatcgccat acagcagaat
     1621 atgttcatga attttcacgc gcagatagtt gcccagcacg ccccacatgt tcttcgcaat
     1681 cagatccaga atgctatggc caatcgcgct gccatacggg ctaatatggc cgaagccctc
     1741 ggggcccagc aggaacagca cggggaacat gccccagctc acgaagaaca gccaggccat
     1801 ggctctcacc agctgtctgc acaggccttt cggcacggta tgaaacgctt caatatacac
     1861 tttggccgca tgaaaatagg tatacatgcc atagctcagg ctaatcagaa aaaacagaat
     1921 tttggtccag ccggtgcaca tcgcgctggt cgcgccccac acaatgcagc ccacgtcgct
     1981 cacctaaaca catgagaaat agagtcataa gcacatgggt cacatacagg agatatgtat
     2041 ataacattaa tacaatttat aaaaaaaagg gggggcacaa ctagtataac ttcgtataag
     2101 gtatactata cgaagttatt aagaggtttc atattgctaa tagcagctac aatccagcta
     2161 ccattctgca taacttcgta taaagtatcc tatacgaagt tatgtcgact ggagagtggc
     2221 cagcccttct catgttcaga gaacatggtt aactggttaa gtcatgtcgt cccacagctg
     2281 cccgtgccct ggcccaccct cgtgaccacc ttcggctacg gcctgcagtg cttcgcccgc
     2341 taccccgacc acatgaagca gcacgacttc ttcaagtccg ccatgcccga aggctacgtc
     2401 caggagcgca ccatcttctt caaggacgac ggcaactaca agacccgcgc cgaggtgaag
     2461 ttcgagggcg acaccctggt gaaccgcatc gagctgaagg gcatcgactt caaggaggac
     2521 ggcaacatcc tggggcacaa gctggagtac aactacaaca gccacaacgt ctatatcatg
     2581 gccgacaagc agaagaacgg catcaaggtg aacttcaaga tccgccacaa catcgaggac
     2641 ggcagcgtgc agctcgccga ccactaccag cagaacaccc ccatcggcga cggccccgtg
     2701 ctgctgcccg acaaccacta cctgagctac cagtccgccc tgagcaaaga ccccaacgag
     2761 aagcgcgatc acatggtcct gctggagttc gtgaccgccg ccgggatcac tctcggcatg
     2821 gacgagctgt acaagtaagc tagcgaagtt cctatacttt ctagagaata ggaacttcta
     2881 agcagacaat cttgtaatac atcagcagct agtagattgc tccagctaga agttcctata
     2941 ccttttgaag aataggaact tcgaattcga tatcaagctt atcgataatc aacctctgga
     3001 ttacaaaatt tgtgaaagat tgactggtat tcttaactat gttgctcctt ttacgctatg
     3061 tggatacgct gctttaatgc ctttgtatca tgctattgct tcccgtatgg ctttcatttt
     3121 ctcctccttg tataaatcct ggttgctgtc tctttatgag gagttgtggc ccgttgtcag
     3181 gcaacgtggc gtggtgtgca ctgtgtttgc tgacgcaacc cccactggtt ggggcattgc
     3241 caccacctgt cagctccttt ccgggacttt cgctttcccc ctccctattg ccacggcgga
     3301 actcatcgcc gcctgccttg cccgctgctg gacaggggct cggctgttgg gcactgacaa
     3361 ttccgtggtg ttgtcgggga aatcatcgtc ctttccttgg ctgctcgcct gtgttgccac
     3421 ctggattctg cgcgggacgt ccttctgcta cgtcccttcg gccctcaatc cagcggacct
     3481 tccttcccgc ggcctgctgc cggctctgcg gcctcttccg cgtcttcgcc ttcgccctca
     3541 gacgagtcgg atctcccttt gggccgcctc cccgcatcga taccg
//
