LOCUS       sequence-265095-        6615 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Intersectional viral expression of BFP in cells expressing both Cre 
            AND Flp.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6615)
  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 6615)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Neuron. 
            2020 Jun 20. pii: S0896-6273(20)30432-3. doi: 
            10.1016/j.neuron.2020.06.003."
COMMENT     Sequence Label: pAAV-Ef1a-Con/Fon-BFP
FEATURES             Location/Qualifiers
     source          1..6615
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..141
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   12..141
                     /label=AAV2 ITR
     promoter        242..1419
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          473..1410
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     1367..1387
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     protein_bind    complement(1426..1459)
                     /label=lox2272
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (AAGTATCC) (Shaw et al., 2021). lox2272 sites are 
                     compatible with each other, but incompatible with loxP or 
                     loxN sites (Lee and Saito, 1988)."
     protein_bind    complement(1510..1543)
                     /label=loxN
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (AGGTATAC) (Shaw et al., 2021). loxN sites are 
                     compatible with each other, but incompatible with loxP or 
                     lox2272 sites."
     protein_bind    1794..1827
                     /label=lox2272
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (AAGTATCC) (Shaw et al., 2021). lox2272 sites are 
                     compatible with each other, but incompatible with loxP or 
                     loxN sites (Lee and Saito, 1988)."
     protein_bind    1878..1911
                     /label=loxN
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (AGGTATAC) (Shaw et al., 2021). loxN sites are 
                     compatible with each other, but incompatible with loxP or 
                     lox2272 sites."
     protein_bind    1918..1951
                     /label=FRT3
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="supports FLP-mediated excision with other FRT3 sites
                     (Bosch et al., 2015)"
     protein_bind    complement(2600..2633)
                     /label=FRT3
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="supports FLP-mediated excision with other FRT3 sites
                     (Bosch et al., 2015)"
     misc_feature    2742..3330
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2795..2815)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3213..3224)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3362..3838
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(3561..3580)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   3878..4018
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3878..4007
                     /label=AAV2 ITR
     rep_origin      4093..4548
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4180..4199)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4390..4411
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4565..4584)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4684..4706
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4744..4762)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4830..4934
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4935..5003,5004..5795)
                     /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(5153..5172)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5966..6554
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6455..6474
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcaaag cccgggcgtc
       61 gggcgacctt tggtcgcccg gcctcagtga gcgagcgagc gcgcagagag ggagtggcca
      121 actccatcac taggggttcc tgcggccgca cgcgtaagct ttgcaaagat ggataaagtt
      181 ttaaacagag aggaatcttt gcagctaatg gaccttctag gtcttgaaag gagtgggaat
      241 tggctccggt gcccgtcagt gggcagagcg cacatcgccc acagtccccg agaagttggg
      301 gggaggggtc ggcaattgaa ccggtgccta gagaaggtgg cgcggggtaa actgggaaag
      361 tgatgtcgtg tactggctcc gcctttttcc cgagggtggg ggagaaccgt atataagtgc
      421 agtagtcgcc gtgaacgttc tttttcgcaa cgggtttgcc gccagaacac aggtaagtgc
      481 cgtgtgtggt tcccgcgggc ctggcctctt tacgggttat ggcccttgcg tgccttgaat
      541 tacttccact ggctgcagta cgtgattctt gatcccgagc ttcgggttgg aagtgggtgg
      601 gagagttcga ggccttgcgc ttaaggagcc ccttcgcctc gtgcttgagt tgaggcctgg
      661 cctgggcgct ggggccgccg cgtgcgaatc tggtggcacc ttcgcgcctg tctcgctgct
      721 ttcgataagt ctctagccat ttaaaatttt tgatgacctg ctgcgacgct ttttttctgg
      781 caagatagtc ttgtaaatgc gggccaagat ctgcacactg gtatttcggt ttttggggcc
      841 gcgggcggcg acggggcccg tgcgtcccag cgcacatgtt cggcgaggcg gggcctgcga
      901 gcgcggccac cgagaatcgg acgggggtag tctcaagctg gccggcctgc tctggtgcct
      961 ggcctcgcgc cgccgtgtat cgccccgccc tgggcggcaa ggctggcccg gtcggcacca
     1021 gttgcgtgag cggaaagatg gccgcttccc ggccctgctg cagggagctc aaaatggagg
     1081 acgcggcgct cgggagagcg ggcgggtgag tcacccacac aaaggaaaag ggcctttccg
     1141 tcctcagccg tcgcttcatg tgactccacg gagtaccggg cgccgtccag gcacctcgat
     1201 tagttctcga gcttttggag tacgtcgtct ttaggttggg gggaggggtt ttatgcgatg
     1261 gagtttcccc acactgagtg ggtggagact gaagttaggc cagcttggca cttgatgtaa
     1321 ttctccttgg aatttgccct ttttgagttt ggatcttggt tcattctcaa gcctcagaca
     1381 gtggttcaaa gtttttttct tccatttcag gtgtcgtgag gatccataac ttcgtatagg
     1441 atactttata cgaagttatg tagctggagc aatttgaaaa gattgttgta ttacctctag
     1501 ctgcgcttaa taacttcgta tagtatacct tatacgaagt tatactagtc ctcctgtact
     1561 cacgggtgtg gttgatgaag gtcttgctgc cgtagaggaa gctagtagcc aggatgtcga
     1621 aggcgaaggg gagagggccg ccctcgacca ccttgattct catggtctgg gtgccctcgt
     1681 agggcttgcc ttcgccctcg gatgtgcact tgaagtgatg gttgtccacg gtgccctcca
     1741 tgtacagctt catgtgcatg ttctccttaa tcagctcgct catggtggct agcataactt
     1801 cgtataaagt atcctatacg aagttatttg ccttaaccca gaaattatca ctgttattct
     1861 ttagataggt gcaaagaata acttcgtata aggtatacta tacgaagtta tggtaccgaa
     1921 gttcctattc ttcaaatagt ataggaactt caattgttat tggtgcaaag aatcacttcc
     1981 ttaagtcttt atactgccca ggaagttcct attcttcaaa aggtatagga acttcggcgc
     2041 gccttaatta agcttgtgcc ccagtttgct agggaggtcg cagtatctgg ccactgccac
     2101 ctcgtgctgc tcgacgtagg tctcgttgtt ggcctccttg attctttcca gtctgtagtc
     2161 cacatagtag acgccaggca tcttgaggtt cttagcgggt ttcttggatc tatatgtggt
     2221 cttgatgttt gcgatcagat ggctcccgcc cacgagcttc agggccatgt cgtttctgcc
     2281 ttccaggccg ccgtcagcgg ggtacagcgt ctcggtgaag gcctcccagc cgagtgtttt
     2341 cttctgcatc acagggccgt tggatgtgaa gttcacccct ctgatcttga cgttgtagat
     2401 gaggcagccg tcctggaggc tggtgtcctg ggtagcggtc agcacgcccc cgtcttcgta
     2461 tgtggtgact ctctcccatg tgaagccctc agggaaggac tgcttgaaga agtcggggat
     2521 gccctctgtg ggacgacatg acttaaccag ttaaccatgt tctctgaaca tgagaagggc
     2581 tggccactct ccagagctcg aagttcctat actatttgaa gaataggaac ttcttgcctt
     2641 aacccagaaa ttatcactgt tattctttag aatggtgcaa agagaagttc ctataccttt
     2701 tgaagaatag gaacttcgaa ttcgatatca agcttatcga taatcaacct ctggattaca
     2761 aaatttgtga aagattgact ggtattctta actatgttgc tccttttacg ctatgtggat
     2821 acgctgcttt aatgcctttg tatcatgcta ttgcttcccg tatggctttc attttctcct
     2881 ccttgtataa atcctggttg ctgtctcttt atgaggagtt gtggcccgtt gtcaggcaac
     2941 gtggcgtggt gtgcactgtg tttgctgacg caacccccac tggttggggc attgccacca
     3001 cctgtcagct cctttccggg actttcgctt tccccctccc tattgccacg gcggaactca
     3061 tcgccgcctg ccttgcccgc tgctggacag gggctcggct gttgggcact gacaattccg
     3121 tggtgttgtc ggggaaatca tcgtcctttc cttggctgct cgcctatgtt gccacctgga
     3181 ttctgcgcgg gacgtccttc tgctacgtcc cttcggccct caatccagcg gaccttcctt
     3241 cccgcggcct gctgccggct ctgcggcctc ttccgcgtct tcgccttcgc cctcagacga
     3301 gtcggatctc cctttgggcc gcctccccgc atcgataccg agcgctgctc gagagatcta
     3361 cgggtggcat ccctgtgacc cctccccagt gcctctcctg gccctggaag ttgccactcc
     3421 agtgcccacc agccttgtcc taataaaatt aagttgcatc attttgtctg actaggtgtc
     3481 cttctataat attatggggt ggaggggggt ggtatggagc aaggggcaag ttgggaagac
     3541 aacctgtagg gcctgcgggg tctattggga accaagctgg agtgcagtgg cacaatcttg
     3601 gctcactgca atctccgcct cctgggttca agcgattctc ctgcctcagc ctcccgagtt
     3661 gttgggattc caggcatgca tgaccaggct cagctaattt ttgttttttt ggtagagacg
     3721 gggtttcacc atattggcca ggctggtctc caactcctaa tctcaggtga tctacccacc
     3781 ttggcctccc aaattgctgg gattacaggc gtgaaccact gctcccttcc ctgtccttct
     3841 gattttgtag gtaaccacgt gcggaccgag cggccgcagg aacccctagt gatggagttg
     3901 gccactccct ctctgcgcgc tcgctcgctc actgaggccg ggcgaccaaa ggtcgcccga
     3961 cgcccgggct ttgcccgggc ggcctcagtg agcgagcgag cgcgcagctg cctgcagggg
     4021 cgcctgatgc ggtattttct ccttacgcat ctgtgcggta tttcacaccg catacgtcaa
     4081 agcaaccata gtacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc
     4141 gcagcgtgac cgctacactt gccagcgccc tagcgcccgc tcctttcgct ttcttccctt
     4201 cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggg ctccctttag
     4261 ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgatttg ggtgatggtt
     4321 cacgtagtgg gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt
     4381 tctttaatag tggactcttg ttccaaactg gaacaacact caaccctatc tcgggctatt
     4441 cttttgattt ataagggatt ttgccgattt cggcctattg gttaaaaaat gagctgattt
     4501 aacaaaaatt taacgcgaat tttaacaaaa tattaacgtt tacaatttta tggtgcactc
     4561 tcagtacaat ctgctctgat gccgcatagt taagccagcc ccgacacccg ccaacacccg
     4621 ctgacgcgcc ctgacgggct tgtctgctcc cggcatccgc ttacagacaa gctgtgaccg
     4681 tctccgggag ctgcatgtgt cagaggtttt caccgtcatc accgaaacgc gcgagacgaa
     4741 agggcctcgt gatacgccta tttttatagg ttaatgtcat gataataatg gtttcttaga
     4801 cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa
     4861 tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt
     4921 gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg
     4981 cattttgcct tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctgaag
     5041 atcagttggg tgcacgagtg ggttacatcg aactggatct caacagcggt aagatccttg
     5101 agagttttcg ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg
     5161 gcgcggtatt atcccgtatt gacgccgggc aagagcaact cggtcgccgc atacactatt
     5221 ctcagaatga cttggttgag tactcaccag tcacagaaaa gcatcttacg gatggcatga
     5281 cagtaagaga attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac
     5341 ttctgacaac gatcggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc
     5401 atgtaactcg ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc
     5461 gtgacaccac gatgcctgta gcaatggcaa caacgttgcg caaactatta actggcgaac
     5521 tacttactct agcttcccgg caacaattaa tagactggat ggaggcggat aaagttgcag
     5581 gaccacttct gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg
     5641 gtgagcgtgg gtctcgcggt atcattgcag cactggggcc agatggtaag ccctcccgta
     5701 tcgtagttat ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg
     5761 ctgagatagg tgcctcactg attaagcatt ggtaactgtc agaccaagtt tactcatata
     5821 tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt
     5881 ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc
     5941 ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct
     6001 tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa
     6061 ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact gtccttctag
     6121 tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc
     6181 tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg
     6241 actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca
     6301 cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat
     6361 gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg
     6421 tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc
     6481 ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc
     6541 ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc
     6601 cttttgctca catgt
//
