LOCUS       sequence-341974-        6898 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Cre-on/Flp-on ChR2-EYFP under the Synapsin promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6898)
  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 6898)
  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 Con/Fon hChR2(H134R)-EYFP
FEATURES             Location/Qualifiers
     source          1..6898
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    441..474
                     /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(570..821)
                     /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(612..633)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     961..980
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     protein_bind    complement(1202..1235)
                     /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(1286..1319)
                     /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     complement(1985..2006)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     protein_bind    2119..2152
                     /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    2203..2236
                     /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      2672..2681
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    complement(2688..2721)
                     /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    2830..3418
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2883..2903)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3301..3312)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3450..3926
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(3649..3668)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   3966..4106
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3966..4095
                     /label=AAV2 ITR
     rep_origin      4181..4636
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4268..4287)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4478..4499
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4653..4672)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4772..4794
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4832..4850)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4918..5022
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5023..5091,5092..5883)
                     /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(5241..5260)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6054..6642
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6543..6562
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   6704..6833
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     promoter        join(6878..6898,1..427)
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
ORIGIN
        1 caggatgagg cggggtgggg gtgcctacct gacgaccgac cccgacccac tggacaagca
       61 cccaaccccc attccccaaa ttgcgcatcc cctatcagag agggggaggg gaaacaggat
      121 gcggcgaggc gcgtgcgcac tgccagcttc agcaccgcgg acagtgcctt cgcccccgcc
      181 tggcggcgcg cgccaccgcc gcctcagcac tgaaggcgcg ctgacgtcac tcgccggtcc
      241 cccgcaaact ccccttcccg gccaccttgg tcgcgtccgc gccgccgccg gcccagccgg
      301 accgcaccac gcgaggcgcg agataggggg gcacgggcgc gaccatctgc gctgcggcgc
      361 cggcgactca gcgctgcctc agtctgcggt gggcagcgga ggagtcgtgt cgtgcctgag
      421 agcgcagtcg agaaggatcc gaagttccta ttcttcaaat agtataggaa cttcgcagag
      481 tagtagctgg attgtagctg ctattagcaa tctaaaacct cttagaagtt cctattcttc
      541 aaaaggtata ggaacttcgg cgcgccttac ttgtacagct cgtccatgcc gagagtgatc
      601 ccggcggcgg tcacgaactc cagcaggacc atgtgatcgc gcttctcgtt ggggtctttg
      661 ctcagggcgg actggtagct caggtagtgg ttgtcgggca gcagcacggg gccgtcgccg
      721 atgggggtgt tctgctggta gtggtcggcg agctgcacgc tgccgtcctc gatgttgtgg
      781 cggatcttga agttcacctt gatgccgttc ttctgcttgt cggccatgat atagacgttg
      841 tggctgttgt agttgtactc cagcttgtgc cccaggatgt tgccgtcctc cttgaagtcg
      901 atgcccttca gctcgatgcg gttcaccagg gtgtcgccct cgaacttcac ctcggcgcgg
      961 gtcttgtagt tgccgtcgtc cttgaagaag atggtgcgct cctggacgta gccttcgggc
     1021 atggcggact tgaagaagtc gtgctgcttc atgtggtcgg ggtagcgggc gaagcactgc
     1081 aggccgtagc cgaaggtggt cacgagggtg ggccagggca cgggcagctg tgggacgaca
     1141 tgacttaacc agttaaccat gttctctgaa catgagaagg gctggccact ctccagtcga
     1201 cataacttcg tataggatac tttatacgaa gttatgcaga atggtagctg gattgtagct
     1261 gctattagca atatgaaacc tcttaataac ttcgtatagt ataccttata cgaagttata
     1321 ctagttgtgc cccccctttt ttttataaat tgtattaatg ttatatacat atctcctgta
     1381 tgtgacccat gtgcttatga ctctatttct catgtgttta ggcctgagca acgactacag
     1441 caggagaacc atgggactcc ttgtctcaga catcgggact atcgtgtggg gggctaccag
     1501 cgccatggca accggctatg ttaaagtcat cttcttttgt cttggattgt gctatggcgc
     1561 gaacacattt tttcacgccg ccaaagcata tatcgagggt tatcatactg tgccaaaggg
     1621 tcggtgccgc caggtcgtga ccggcatggc atggctgttt ttcgtgagct ggggtatgtt
     1681 cccaattctc ttcattttgg ggcccgaagg ttttggcgtc ctgagcgtct atggctccac
     1741 cgtaggtcac acgattattg atctgatgag taaaaattgt tgggggttgt tgggacacta
     1801 cctgcgcgtc ctgatccacg agcacatatt gattcacgga gatatccgca aaaccaccaa
     1861 actgaacatc ggcggaacgg agatcgaggt cgagactctc gtcgaagacg aagccgaggc
     1921 cggagccgtg ccagcggccg ccgtgagcaa gggcgaggag ctgttcaccg gggtggtgcc
     1981 catcctggtc gagctggacg gcgacgtaaa cggccacaag ttcagcgtgt ccggcgaggg
     2041 cgagggcgat gccacctacg gcaagctgac cctgaagttc atctgcacca ccggcaaggt
     2101 gagtacagga ggggtaccat aacttcgtat aaagtatcct atacgaagtt atttgcctta
     2161 acccagaaat tatcactgtt attctttaga atggtgcaaa gaataacttc gtataaggta
     2221 tactatacga agttatgcta gcaaccccga cacttaccgg tgaggttgct caggcggata
     2281 aggatgacag gacaagtgag cagccactct gcatagcgca gccactgcac ccggtgtcct
     2341 gtggcaaggt agagcataga gggattctta aactcaaaaa agaactcgag aatcacctta
     2401 accatttcaa tggcgcacac atagatctcc tcccagccgc atgtagattt ccaggtttgg
     2461 taggcataga acatcagcag caaaatgctg aatcctgctg caagccactg caggacattt
     2521 gacgcggtct gagcgccgtt cgtgccgcga gattcaatcc atccggcaca gtaacattga
     2581 tcctcaggga ccaggacgga cccgttcacc accacaggat tagtaacgaa caaaagttcg
     2641 cgtccgacgg cagacaaagc gccgccatag tccatggtgg cgagctcgaa gttcctatac
     2701 tatttgaaga ataggaactt cttgccttaa cccagaaatt atcactgtta ttctttagaa
     2761 tggtgcaaag agaagttcct ataccttttg aagaatagga acttcgaatt cgatatcaag
     2821 cttatcgata atcaacctct ggattacaaa atttgtgaaa gattgactgg tattcttaac
     2881 tatgttgctc cttttacgct atgtggatac gctgctttaa tgcctttgta tcatgctatt
     2941 gcttcccgta tggctttcat tttctcctcc ttgtataaat cctggttgct gtctctttat
     3001 gaggagttgt ggcccgttgt caggcaacgt ggcgtggtgt gcactgtgtt tgctgacgca
     3061 acccccactg gttggggcat tgccaccacc tgtcagctcc tttccgggac tttcgctttc
     3121 cccctcccta ttgccacggc ggaactcatc gccgcctgcc ttgcccgctg ctggacaggg
     3181 gctcggctgt tgggcactga caattccgtg gtgttgtcgg ggaaatcatc gtcctttcct
     3241 tggctgctcg cctatgttgc cacctggatt ctgcgcggga cgtccttctg ctacgtccct
     3301 tcggccctca atccagcgga ccttccttcc cgcggcctgc tgccggctct gcggcctctt
     3361 ccgcgtcttc gccttcgccc tcagacgagt cggatctccc tttgggccgc ctccccgcat
     3421 cgataccgag cgctgctcga gagatctacg ggtggcatcc ctgtgacccc tccccagtgc
     3481 ctctcctggc cctggaagtt gccactccag tgcccaccag ccttgtccta ataaaattaa
     3541 gttgcatcat tttgtctgac taggtgtcct tctataatat tatggggtgg aggggggtgg
     3601 tatggagcaa ggggcaagtt gggaagacaa cctgtagggc ctgcggggtc tattgggaac
     3661 caagctggag tgcagtggca caatcttggc tcactgcaat ctccgcctcc tgggttcaag
     3721 cgattctcct gcctcagcct cccgagttgt tgggattcca ggcatgcatg accaggctca
     3781 gctaattttt gtttttttgg tagagacggg gtttcaccat attggccagg ctggtctcca
     3841 actcctaatc tcaggtgatc tacccacctt ggcctcccaa attgctggga ttacaggcgt
     3901 gaaccactgc tcccttccct gtccttctga ttttgtaggt aaccacgtgc ggaccgagcg
     3961 gccgcaggaa cccctagtga tggagttggc cactccctct ctgcgcgctc gctcgctcac
     4021 tgaggccggg cgaccaaagg tcgcccgacg cccgggcttt gcccgggcgg cctcagtgag
     4081 cgagcgagcg cgcagctgcc tgcaggggcg cctgatgcgg tattttctcc ttacgcatct
     4141 gtgcggtatt tcacaccgca tacgtcaaag caaccatagt acgcgccctg tagcggcgca
     4201 ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcctta
     4261 gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt
     4321 caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg gcacctcgac
     4381 cccaaaaaac ttgatttggg tgatggttca cgtagtgggc catcgccctg atagacggtt
     4441 tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga
     4501 acaacactca actctatctc gggctattct tttgatttat aagggatttt gccgatttcg
     4561 gtctattggt taaaaaatga gctgatttaa caaaaattta acgcgaattt taacaaaata
     4621 ttaacgttta caattttatg gtgcactctc agtacaatct gctctgatgc cgcatagtta
     4681 agccagcccc gacacccgcc aacacccgct gacgcgccct gacgggcttg tctgctcccg
     4741 gcatccgctt acagacaagc tgtgaccgtc tccgggagct gcatgtgtca gaggttttca
     4801 ccgtcatcac cgaaacgcgc gagacgaaag ggcctcgtga tacgcctatt tttataggtt
     4861 aatgtcatga taataatggt ttcttagacg tcaggtggca cttttcgggg aaatgtgcgc
     4921 ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa
     4981 taaccctgat aaatgcttca ataatattga aaaaggaaga gtatgagtat tcaacatttc
     5041 cgtgtcgccc ttattccctt ttttgcggca ttttgccttc ctgtttttgc tcacccagaa
     5101 acgctggtga aagtaaaaga tgctgaagat cagttgggtg cacgagtggg ttacatcgaa
     5161 ctggatctca acagcggtaa gatccttgag agttttcgcc ccgaagaacg ttttccaatg
     5221 atgagcactt ttaaagttct gctatgtggc gcggtattat cccgtattga cgccgggcaa
     5281 gagcaactcg gtcgccgcat acactattct cagaatgact tggttgagta ctcaccagtc
     5341 acagaaaagc atcttacgga tggcatgaca gtaagagaat tatgcagtgc tgccataacc
     5401 atgagtgata acactgcggc caacttactt ctgacaacga tcggaggacc gaaggagcta
     5461 accgcttttt tgcacaacat gggggatcat gtaactcgcc ttgatcgttg ggaaccggag
     5521 ctgaatgaag ccataccaaa cgacgagcgt gacaccacga tgcctgtagc aatggcaaca
     5581 acgttgcgca aactattaac tggcgaacta cttactctag cttcccggca acaattaata
     5641 gactggatgg aggcggataa agttgcagga ccacttctgc gctcggccct tccggctggc
     5701 tggtttattg ctgataaatc tggagccggt gagcgtgggt ctcgcggtat cattgcagca
     5761 ctggggccag atggtaagcc ctcccgtatc gtagttatct acacgacggg gagtcaggca
     5821 actatggatg aacgaaatag acagatcgct gagataggtg cctcactgat taagcattgg
     5881 taactgtcag accaagttta ctcatatata ctttagattg atttaaaact tcatttttaa
     5941 tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt
     6001 gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat
     6061 cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg
     6121 gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga
     6181 gcgcagatac caaatactgt tcttctagtg tagccgtagt taggccacca cttcaagaac
     6241 tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt
     6301 ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag
     6361 cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc
     6421 gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag
     6481 gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca
     6541 gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt
     6601 cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc
     6661 tttttacggt tcctggcctt ttgctggcct tttgctcaca tgtcctgcag gcagctgcgc
     6721 gctcgctcgc tcactgaggc cgcccgggcg tcgggcgacc tttggtcgcc cggcctcagt
     6781 gagcgagcga gcgcgcagag agggagtggc caactccatc actaggggtt cctgcggccg
     6841 cacgcgtgtg tctagactgc agagggccct gcgtatgagt gcaagtgggt tttaggac
//
