LOCUS       sequence-271932-        6616 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Intersectional viral expression of mCherry in cells expressing Cre 
            AND NOT Flp.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6616)
  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 6616)
  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/Foff 2.0-mCherry
FEATURES             Location/Qualifiers
     source          1..6616
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   6..135
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     promoter        236..1414
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          467..1405
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     1362..1382
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     protein_bind    complement(1421..1454)
                     /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(1505..1538)
                     /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."
     primer_bind     1737..1755
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     regulatory      1897..1906
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    1913..1946
                     /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    1997..2030
                     /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    2037..2070
                     /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     2302..2321
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     protein_bind    complement(2606..2639)
                     /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    2748..3336
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2801..2821)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3219..3230)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3368..3844
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(3567..3586)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   3884..4024
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3884..4013
                     /label=AAV2 ITR
     rep_origin      4099..4554
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4186..4205)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4396..4417
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4571..4590)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4690..4712
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4750..4768)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4836..4940
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4941..5009,5010..5801)
                     /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(5159..5178)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5972..6560
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6461..6480
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 catgtcctgc aggcagctgc gcgctcgctc gctcactgag gccgcccggg cgtcgggcga
       61 cctttggtcg cccggcctca gtgagcgagc gagcgcgcag agagggagtg gccaactcca
      121 tcactagggg ttcctgcggc cgcacgcgta agctttgcaa agatggataa agttttaaac
      181 agagaggaat ctttgcagct aatggacctt ctaggtcttg aaaggagtgg gaattggctc
      241 cggtgcccgt cagtgggcag agcgcacatc gcccacagtc cccgagaagt tggggggagg
      301 ggtcggcaat tgaaccggtg cctagagaag gtggcgcggg gtaaactggg aaagtgatgt
      361 cgtgtactgg ctccgccttt ttcccgaggg tgggggagaa ccgtatataa gtgcagtagt
      421 cgccgtgaac gttctttttc gcaacgggtt tgccgccaga acacaggtaa gtgccgtgtg
      481 tggttcccgc gggcctggcc tctttacggg ttatggccct tgcgtgcctt gaattacttc
      541 cacctggctg cagtacgtga ttcttgatcc cgagcttcgg gttggaagtg ggtgggagag
      601 ttcgaggcct tgcgcttaag gagccccttc gcctcgtgct tgagttgagg cctggcctgg
      661 gcgctggggc cgccgcgtgc gaatctggtg gcaccttcgc gcctgtctcg ctgctttcga
      721 taagtctcta gccatttaaa atttttgatg acctgctgcg acgctttttt tctggcaaga
      781 tagtcttgta aatgcgggcc aagatctgca cactggtatt tcggtttttg gggccgcggg
      841 cggcgacggg gcccgtgcgt cccagcgcac atgttcggcg aggcggggcc tgcgagcgcg
      901 gccaccgaga atcggacggg ggtagtctca agctggccgg cctgctctgg tgcctggcct
      961 cgcgccgccg tgtatcgccc cgccctgggc ggcaaggctg gcccggtcgg caccagttgc
     1021 gtgagcggaa agatggccgc ttcccggccc tgctgcaggg agctcaaaat ggaggacgcg
     1081 gcgctcggga gagcgggcgg gtgagtcacc cacacaaagg aaaagggcct ttccgtcctc
     1141 agccgtcgct tcatgtgact ccacggagta ccgggcgccg tccaggcacc tcgattagtt
     1201 ctcgagcttt tggagtacgt cgtctttagg ttggggggag gggttttatg cgatggagtt
     1261 tccccacact gagtgggtgg agactgaagt taggccagct tggcacttga tgtaattctc
     1321 cttggaattt gccctttttg agtttggatc ttggttcatt ctcaagcctc agacagtggt
     1381 tcaaagtttt tttcttccat ttcaggtgtc gtgaggatcc ataacttcgt ataggatact
     1441 ttatacgaag ttatgtagct ggagcaattt gaaaagattg ttgtattacc tctagctgcg
     1501 cttaataact tcgtatagta taccttatac gaagttatgt cgaccctcct gtactcacct
     1561 gggtcacggt caccacgccg ccgtcctcga agttcatcac gcgctcccac ttgaagccct
     1621 cggggaagga cagcttcaag tagtcgggga tgtcggcggg gtgcttcacg taggccttgg
     1681 agccgtacat gaactgaggg gacaggatgt cccaggcgaa gggcaggggg ccacccttgg
     1741 tcaccttcag cttggcggtc tgggtgccct cgtaggggcg gccctcgccc tcgccctcga
     1801 tctcgaactc gtggccgttc acggagccct ccatgtgcac cttgaagcgc atgaactcct
     1861 tgatgatggc catgttatcc tcctcgccct tgctcaccat ggtggcacta gtataacttc
     1921 gtataaagta tcctatacga agttatttgc cttaacccag aaattatcac tgttattctt
     1981 tagataggtg caaagaataa cttcgtataa ggtatactat acgaagttat ggtaccgaag
     2041 ttcctattct ctagaaagta taggaacttc gcagaatggt agctggattg tagctgctat
     2101 tagcaatatg aaacctctta gaagttccta ttcttcaaaa ggtataggaa cttcggcgcg
     2161 cctggagagt ggccagccct tctcatgttc agagaacatg gttaactggt taagtcatgt
     2221 cgtcccacag gactcctccc tgcaggacgg cgagttcatc tacaaggtga agctgcgcgg
     2281 caccaacttc ccctccgacg gccccgtaat gcagaagaag accatgggct gggaggcctc
     2341 ctccgagcgg atgtaccccg aggacggcgc cctgaagggc gagatcaagc agaggctgaa
     2401 gctgaaggac ggcggccact acgacgctga ggtcaagacc acctacaagg ccaagaagcc
     2461 cgtgcagctg cccggcgcct acaacgtcaa catcaagttg gacatcacct cccacaacga
     2521 ggactacacc atcgtggaac agtacgaacg cgccgagggc cgccactcca ccggcggcat
     2581 ggacgagctg tacaagtaag ctagcgaagt tcctatactt tctagagaat aggaacttct
     2641 aagcagacaa tcttgtaata catcagcagc tagtagattg ctccagctag aagttcctat
     2701 accttttgaa gaataggaac ttcgaattcg atatcaagct tatcgataat caacctctgg
     2761 attacaaaat ttgtgaaaga ttgactggta ttcttaacta tgttgctcct tttacgctat
     2821 gtggatacgc tgctttaatg cctttgtatc atgctattgc ttcccgtatg gctttcattt
     2881 tctcctcctt gtataaatcc tggttgctgt ctctttatga ggagttgtgg cccgttgtca
     2941 ggcaacgtgg cgtggtgtgc actgtgtttg ctgacgcaac ccccactggt tggggcattg
     3001 ccaccacctg tcagctcctt tccgggactt tcgctttccc cctccctatt gccacggcgg
     3061 aactcatcgc cgcctgcctt gcccgctgct ggacaggggc tcggctgttg ggcactgaca
     3121 attccgtggt gttgtcgggg aaatcatcgt cctttccttg gctgctcgcc tatgttgcca
     3181 cctggattct gcgcgggacg tccttctgct acgtcccttc ggccctcaat ccagcggacc
     3241 ttccttcccg cggcctgctg ccggctctgc ggcctcttcc gcgtcttcgc cttcgccctc
     3301 agacgagtcg gatctccctt tgggccgcct ccccgcatcg ataccgagcg ctgctcgaga
     3361 gatctacggg tggcatccct gtgacccctc cccagtgcct ctcctggccc tggaagttgc
     3421 cactccagtg cccaccagcc ttgtcctaat aaaattaagt tgcatcattt tgtctgacta
     3481 ggtgtccttc tataatatta tggggtggag gggggtggta tggagcaagg ggcaagttgg
     3541 gaagacaacc tgtagggcct gcggggtcta ttgggaacca agctggagtg cagtggcaca
     3601 atcttggctc actgcaatct ccgcctcctg ggttcaagcg attctcctgc ctcagcctcc
     3661 cgagttgttg ggattccagg catgcatgac caggctcagc taatttttgt ttttttggta
     3721 gagacggggt ttcaccatat tggccaggct ggtctccaac tcctaatctc aggtgatcta
     3781 cccaccttgg cctcccaaat tgctgggatt acaggcgtga accactgctc ccttccctgt
     3841 ccttctgatt ttgtaggtaa ccacgtgcgg accgagcggc cgcaggaacc cctagtgatg
     3901 gagttggcca ctccctctct gcgcgctcgc tcgctcactg aggccgggcg accaaaggtc
     3961 gcccgacgcc cgggctttgc ccgggcggcc tcagtgagcg agcgagcgcg cagctgcctg
     4021 caggggcgcc tgatgcggta ttttctcctt acgcatctgt gcggtatttc acaccgcata
     4081 cgtcaaagca accatagtac gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg
     4141 ttacgcgcag cgtgaccgct acacttgcca gcgccttagc gcccgctcct ttcgctttct
     4201 tcccttcctt tctcgccacg ttcgccggct ttccccgtca agctctaaat cgggggctcc
     4261 ctttagggtt ccgatttagt gctttacggc acctcgaccc caaaaaactt gatttgggtg
     4321 atggttcacg tagtgggcca tcgccctgat agacggtttt tcgccctttg acgttggagt
     4381 ccacgttctt taatagtgga ctcttgttcc aaactggaac aacactcaac tctatctcgg
     4441 gctattcttt tgatttataa gggattttgc cgatttcggt ctattggtta aaaaatgagc
     4501 tgatttaaca aaaatttaac gcgaatttta acaaaatatt aacgtttaca attttatggt
     4561 gcactctcag tacaatctgc tctgatgccg catagttaag ccagccccga cacccgccaa
     4621 cacccgctga cgcgccctga cgggcttgtc tgctcccggc atccgcttac agacaagctg
     4681 tgaccgtctc cgggagctgc atgtgtcaga ggttttcacc gtcatcaccg aaacgcgcga
     4741 gacgaaaggg cctcgtgata cgcctatttt tataggttaa tgtcatgata ataatggttt
     4801 cttagacgtc aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt
     4861 tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat
     4921 aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt
     4981 ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg
     5041 ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga
     5101 tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc
     5161 tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga gcaactcggt cgccgcatac
     5221 actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg
     5281 gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca
     5341 acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg
     5401 gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg
     5461 acgagcgtga caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg
     5521 gcgaactact tactctagct tcccggcaac aattaataga ctggatggag gcggataaag
     5581 ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg
     5641 gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct
     5701 cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac
     5761 agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac caagtttact
     5821 catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga
     5881 tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt
     5941 cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct
     6001 gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc
     6061 taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc
     6121 ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc
     6181 tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg
     6241 ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt
     6301 cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg
     6361 agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg
     6421 gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt
     6481 atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag
     6541 gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt
     6601 gctggccttt tgctca
//
