LOCUS       sequence-86451-e        6909 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Cre-off/Flp-on ChR2-EYFP under the Synapsin promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6909)
  AUTHORS   Fenno LE, Mattis J, Ramakrishnan C, Hyun M, Lee SY, He M, Tucciarone
            J, Selimbeyoglu A, Berndt A, Grosenick L, Zalocusky KA, Bernstein H,
            Swanson H, Perry C, Diester I, Boyce FM, Bass CE, Neve R, Huang ZJ, 
            Deisseroth K
  TITLE     Targeting cells with single vectors using multiple-feature Boolean 
            logic.
  JOURNAL   Nat Methods. 2014 Jul;11(7):763-72. doi: 10.1038/nmeth.2996. Epub 
            2014 Jun 8.
  PUBMED    24908100
REFERENCE   2  (bases 1 to 6909)
  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"; doi: 
            "10.1038/nmeth.2996"; journalName: "Nat Methods"; date: "2014-07"; 
            volume: "11"; issue: "7"; pages: "763-72"
COMMENT     Sequence Label: pAAV-hSyn Coff/Fon hChR2(H134R)-EYFP
FEATURES             Location/Qualifiers
     source          1..6909
                     /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        186..633
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     protein_bind    647..680
                     /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)"
     CDS             complement(776..1027)
                     /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     complement(818..839)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     1167..1186
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     protein_bind    complement(1408..1441)
                     /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(1492..1525)
                     /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     1638..1659
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     protein_bind    2325..2358
                     /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    2409..2442
                     /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."
     regulatory      2878..2887
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    complement(2894..2927)
                     /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    3036..3624
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3089..3109)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3507..3518)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3656..4132
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(3855..3874)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   4172..4312
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4172..4301
                     /label=AAV2 ITR
     rep_origin      4387..4842
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4474..4493)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4684..4705
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4859..4878)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4978..5000
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5038..5056)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5124..5228
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5229..5297,5298..6089)
                     /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(5447..5466)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6260..6848
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6749..6768
                     /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 cgcgtgtgtc tagactgcag agggccctgc
      181 gtatgagtgc aagtgggttt taggaccagg atgaggcggg gtgggggtgc ctacctgacg
      241 accgaccccg acccactgga caagcaccca acccccattc cccaaattgc gcatccccta
      301 tcagagaggg ggaggggaaa caggatgcgg cgaggcgcgt gcgcactgcc agcttcagca
      361 ccgcggacag tgccttcgcc cccgcctggc ggcgcgcgcc accgccgcct cagcactgaa
      421 ggcgcgctga cgtcactcgc cggtcccccg caaactcccc ttcccggcca ccttggtcgc
      481 gtccgcgccg ccgccggccc agccggaccg caccacgcga ggcgcgagat aggggggcac
      541 gggcgcgacc atctgcgctg cggcgccggc gactcagcgc tgcctcagtc tgcggtgggc
      601 agcggaggag tcgtgtcgtg cctgagagcg cagtcgagaa ggatccgaag ttcctattct
      661 tcaaatagta taggaacttc gcagagtagt agctggattg tagctgctat tagcaatcta
      721 aaacctctta gaagttccta ttcttcaaaa ggtataggaa cttcggcgcg ccttacttgt
      781 acagctcgtc catgccgaga gtgatcccgg cggcggtcac gaactccagc aggaccatgt
      841 gatcgcgctt ctcgttgggg tctttgctca gggcggactg gtagctcagg tagtggttgt
      901 cgggcagcag cacggggccg tcgccgatgg gggtgttctg ctggtagtgg tcggcgagct
      961 gcacgctgcc gtcctcgatg ttgtggcgga tcttgaagtt caccttgatg ccgttcttct
     1021 gcttgtcggc catgatatag acgttgtggc tgttgtagtt gtactccagc ttgtgcccca
     1081 ggatgttgcc gtcctccttg aagtcgatgc ccttcagctc gatgcggttc accagggtgt
     1141 cgccctcgaa cttcacctcg gcgcgggtct tgtagttgcc gtcgtccttg aagaagatgg
     1201 tgcgctcctg gacgtagcct tcgggcatgg cggacttgaa gaagtcgtgc tgcttcatgt
     1261 ggtcggggta gcgggcgaag cactgcaggc cgtagccgaa ggtggtcacg agggtgggcc
     1321 agggcacggg cagctgtggg acgacatgac ttaaccagtt aaccatgttc tctgaacatg
     1381 agaagggctg gccactctcc agtcgacata acttcgtata ggatacttta tacgaagtta
     1441 tgcagaatgg tagctggatt gtagctgcta ttagcaatat gaaacctctt aataacttcg
     1501 tatagtatac cttatacgaa gttatactag tcctcctgta ctcaccttgc cggtggtgca
     1561 gatgaacttc agggtcagct tgccgtaggt ggcatcgccc tcgccctcgc cggacacgct
     1621 gaacttgtgg ccgtttacgt cgccgtccag ctcgaccagg atgggcacca ccccggtgaa
     1681 cagctcctcg cccttgctca cggcggccgc tggcacggct ccggcctcgg cttcgtcttc
     1741 gacgagagtc tcgacctcga tctccgttcc gccgatgttc agtttggtgg ttttgcggat
     1801 atctccgtga atcaatatgt gctcgtggat caggacgcgc aggtagtgtc ccaacaaccc
     1861 ccaacaattt ttactcatca gatcaataat cgtgtgacct acggtggagc catagacgct
     1921 caggacgcca aaaccttcgg gccccaaaat gaagagaatt gggaacatac cccagctcac
     1981 gaaaaacagc catgccatgc cggtcacgac ctggcggcac cgaccctttg gcacagtatg
     2041 ataaccctcg atatatgctt tggcggcgtg aaaaaatgtg ttcgcgccat agcacaatcc
     2101 aagacaaaag aagatgactt taacatagcc ggttgccatg gcgctggtag ccccccacac
     2161 gatagtcccg atgtctgaga caaggagtcc catggttctc ctgctgtagt cgttgctcag
     2221 gcctaaacac atgagaaata gagtcataag cacatgggtc acatacagga gatatgtata
     2281 taacattaat acaatttata aaaaaaaggg ggggcacagg taccataact tcgtataaag
     2341 tatcctatac gaagttattt gccttaaccc agaaattatc actgttattc tttagaatgg
     2401 tgcaaagaat aacttcgtat aaggtatact atacgaagtt atgctagcaa ccccgacact
     2461 taccggtgag gttgctcagg cggataagga tgacaggaca agtgagcagc cactctgcat
     2521 agcgcagcca ctgcacccgg tgtcctgtgg caaggtagag catagaggga ttcttaaact
     2581 caaaaaagaa ctcgagaatc accttaacca tttcaatggc gcacacatag atctcctccc
     2641 agccgcatgt agatttccag gtttggtagg catagaacat cagcagcaaa atgctgaatc
     2701 ctgctgcaag ccactgcagg acatttgacg cggtctgagc gccgttcgtg ccgcgagatt
     2761 caatccatcc ggcacagtaa cattgatcct cagggaccag gacggacccg ttcaccacca
     2821 caggattagt aacgaacaaa agttcgcgtc cgacggcaga caaagcgccg ccatagtcca
     2881 tggtggcgag ctcgaagttc ctatactatt tgaagaatag gaacttcttg ccttaaccca
     2941 gaaattatca ctgttattct ttagaatggt gcaaagagaa gttcctatac cttttgaaga
     3001 ataggaactt cgaattcgat atcaagctta tcgataatca acctctggat tacaaaattt
     3061 gtgaaagatt gactggtatt cttaactatg ttgctccttt tacgctatgt ggatacgctg
     3121 ctttaatgcc tttgtatcat gctattgctt cccgtatggc tttcattttc tcctccttgt
     3181 ataaatcctg gttgctgtct ctttatgagg agttgtggcc cgttgtcagg caacgtggcg
     3241 tggtgtgcac tgtgtttgct gacgcaaccc ccactggttg gggcattgcc accacctgtc
     3301 agctcctttc cgggactttc gctttccccc tccctattgc cacggcggaa ctcatcgccg
     3361 cctgccttgc ccgctgctgg acaggggctc ggctgttggg cactgacaat tccgtggtgt
     3421 tgtcggggaa atcatcgtcc tttccttggc tgctcgcctg tgttgccacc tggattctgc
     3481 gcgggacgtc cttctgctac gtcccttcgg ccctcaatcc agcggacctt ccttcccgcg
     3541 gcctgctgcc ggctctgcgg cctcttccgc gtcttcgcct tcgccctcag acgagtcgga
     3601 tctccctttg ggccgcctcc ccgcatcgat accgagcgct gctcgagaga tctacgggtg
     3661 gcatccctgt gacccctccc cagtgcctct cctggccctg gaagttgcca ctccagtgcc
     3721 caccagcctt gtcctaataa aattaagttg catcattttg tctgactagg tgtccttcta
     3781 taatattatg gggtggaggg gggtggtatg gagcaagggg caagttggga agacaacctg
     3841 tagggcctgc ggggtctatt gggaaccaag ctggagtgca gtggcacaat cttggctcac
     3901 tgcaatctcc gcctcctggg ttcaagcgat tctcctgcct cagcctcccg agttgttggg
     3961 attccaggca tgcatgacca ggctcagcta atttttgttt ttttggtaga gacggggttt
     4021 caccatattg gccaggctgg tctccaactc ctaatctcag gtgatctacc caccttggcc
     4081 tcccaaattg ctgggattac aggcgtgaac cactgctccc ttccctgtcc ttctgatttt
     4141 gtaggtaacc acgtgcggac cgagcggccg caggaacccc tagtgatgga gttggccact
     4201 ccctctctgc gcgctcgctc gctcactgag gccgggcgac caaaggtcgc ccgacgcccg
     4261 ggctttgccc gggcggcctc agtgagcgag cgagcgcgca gctgcctgca ggggcgcctg
     4321 atgcggtatt ttctccttac gcatctgtgc ggtatttcac accgcatacg tcaaagcaac
     4381 catagtacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg
     4441 tgaccgctac acttgccagc gccctagcgc ccgctccttt cgctttcttc ccttcctttc
     4501 tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg ggggctccct ttagggttcc
     4561 gatttagtgc tttacggcac ctcgacccca aaaaacttga tttgggtgat ggttcacgta
     4621 gtgggccatc gccctgatag acggtttttc gccctttgac gttggagtcc acgttcttta
     4681 atagtggact cttgttccaa actggaacaa cactcaaccc tatctcgggc tattcttttg
     4741 atttataagg gattttgccg atttcggcct attggttaaa aaatgagctg atttaacaaa
     4801 aatttaacgc gaattttaac aaaatattaa cgtttacaat tttatggtgc actctcagta
     4861 caatctgctc tgatgccgca tagttaagcc agccccgaca cccgccaaca cccgctgacg
     4921 cgccctgacg ggcttgtctg ctcccggcat ccgcttacag acaagctgtg accgtctccg
     4981 ggagctgcat gtgtcagagg ttttcaccgt catcaccgaa acgcgcgaga cgaaagggcc
     5041 tcgtgatacg cctattttta taggttaatg tcatgataat aatggtttct tagacgtcag
     5101 gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt
     5161 caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa
     5221 ggaagagtat gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt
     5281 gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt
     5341 tgggtgcacg agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt
     5401 ttcgccccga agaacgtttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg
     5461 tattatcccg tattgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga
     5521 atgacttggt tgagtactca ccagtcacag aaaagcatct tacggatggc atgacagtaa
     5581 gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga
     5641 caacgatcgg aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa
     5701 ctcgccttga tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca
     5761 ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact attaactggc gaactactta
     5821 ctctagcttc ccggcaacaa ttaatagact ggatggaggc ggataaagtt gcaggaccac
     5881 ttctgcgctc ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc
     5941 gtgggtctcg cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag
     6001 ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga
     6061 taggtgcctc actgattaag cattggtaac tgtcagacca agtttactca tatatacttt
     6121 agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata
     6181 atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag
     6241 aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa
     6301 caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt
     6361 ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgtcctt ctagtgtagc
     6421 cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa
     6481 tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa
     6541 gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc
     6601 ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa
     6661 gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa
     6721 caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg
     6781 ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc
     6841 tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg
     6901 ctcacatgt
//
