LOCUS       Exported                6898 bp ds-DNA     circular SYN 08-SEP-2021
DEFINITION  Cre-on/Flp-on ChR2-EYFP under the Synapsin promoter.
ACCESSION   .
VERSION     .
KEYWORDS    pAAV-hSyn Con/Fon hChR2(H134R)-EYFP
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 Sep 8, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6898
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     repeat_region   66..195
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     rep_origin      complement(257..845)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(337..356)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(1016..1876)
                     /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     1639..1658
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1877..1981)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     2049..2067
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(2105..2127)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     2227..2246
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(2263..2718)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2400..2421)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2612..2631
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     repeat_region   2793..2933
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2804..2933
                     /label=AAV2 ITR
     polyA_signal    2973..3449
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     3231..3250
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     misc_feature    3481..4069
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     CDS             3587..3598
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     3996..4016
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     regulatory      4218..4227
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     protein_bind    4663..4696
                     /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    4747..4780
                     /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     4893..4914
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     protein_bind    complement(5580..5613)
                     /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(5664..5697)
                     /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(5919..5938)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     CDS             6078..6329
                     /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     6266..6287
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     promoter        complement(6472..21)
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
ORIGIN
        1 gtcctaaaac ccacttgcac tcatacgcag ggccctctgc agtctagaca cacgcgtgcg
       61 gccgcaggaa cccctagtga tggagttggc cactccctct ctgcgcgctc gctcgctcac
      121 tgaggccggg cgaccaaagg tcgcccgacg cccgggcggc ctcagtgagc gagcgagcgc
      181 gcagctgcct gcaggacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg
      241 ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac
      301 gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg
      361 gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct
      421 ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg
      481 tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct
      541 gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac
      601 tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt
      661 tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc
      721 tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca
      781 ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat
      841 ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac
      901 gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt
      961 aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc
     1021 aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg
     1081 cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg
     1141 ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc
     1201 cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta
     1261 ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg
     1321 ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct
     1381 ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta
     1441 gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg
     1501 ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga
     1561 ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt
     1621 gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca
     1681 ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt
     1741 cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt
     1801 ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga
     1861 aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat cagggttatt
     1921 gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc
     1981 gcacatttcc ccgaaaagtg ccacctgacg tctaagaaac cattattatc atgacattaa
     2041 cctataaaaa taggcgtatc acgaggccct ttcgtctcgc gcgtttcggt gatgacggtg
     2101 aaaacctctg acacatgcag ctcccggaga cggtcacagc ttgtctgtaa gcggatgccg
     2161 ggagcagaca agcccgtcag ggcgcgtcag cgggtgttgg cgggtgtcgg ggctggctta
     2221 actatgcggc atcagagcag attgtactga gagtgcacca taaaattgta aacgttaata
     2281 ttttgttaaa attcgcgtta aatttttgtt aaatcagctc attttttaac caatagaccg
     2341 aaatcggcaa aatcccttat aaatcaaaag aatagcccga gatagagttg agtgttgttc
     2401 cagtttggaa caagagtcca ctattaaaga acgtggactc caacgtcaaa gggcgaaaaa
     2461 ccgtctatca gggcgatggc ccactacgtg aaccatcacc caaatcaagt tttttggggt
     2521 cgaggtgccg taaagcacta aatcggaacc ctaaagggag cccccgattt agagcttgac
     2581 ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa agcgaaagga gcgggcgcta
     2641 aggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac cacacccgcc gcgcttaatg
     2701 cgccgctaca gggcgcgtac tatggttgct ttgacgtatg cggtgtgaaa taccgcacag
     2761 atgcgtaagg agaaaatacc gcatcaggcg cccctgcagg cagctgcgcg ctcgctcgct
     2821 cactgaggcc gcccgggcaa agcccgggcg tcgggcgacc tttggtcgcc cggcctcagt
     2881 gagcgagcga gcgcgcagag agggagtggc caactccatc actaggggtt cctgcggccg
     2941 ctcggtccgc acgtggttac ctacaaaatc agaaggacag ggaagggagc agtggttcac
     3001 gcctgtaatc ccagcaattt gggaggccaa ggtgggtaga tcacctgaga ttaggagttg
     3061 gagaccagcc tggccaatat ggtgaaaccc cgtctctacc aaaaaaacaa aaattagctg
     3121 agcctggtca tgcatgcctg gaatcccaac aactcgggag gctgaggcag gagaatcgct
     3181 tgaacccagg aggcggagat tgcagtgagc caagattgtg ccactgcact ccagcttggt
     3241 tcccaataga ccccgcaggc cctacaggtt gtcttcccaa cttgcccctt gctccatacc
     3301 acccccctcc accccataat attatagaag gacacctagt cagacaaaat gatgcaactt
     3361 aattttatta ggacaaggct ggtgggcact ggagtggcaa cttccagggc caggagaggc
     3421 actggggagg ggtcacaggg atgccacccg tagatctctc gagcagcgct cggtatcgat
     3481 gcggggaggc ggcccaaagg gagatccgac tcgtctgagg gcgaaggcga agacgcggaa
     3541 gaggccgcag agccggcagc aggccgcggg aaggaaggtc cgctggattg agggccgaag
     3601 ggacgtagca gaaggacgtc ccgcgcagaa tccaggtggc aacataggcg agcagccaag
     3661 gaaaggacga tgatttcccc gacaacacca cggaattgtc agtgcccaac agccgagccc
     3721 ctgtccagca gcgggcaagg caggcggcga tgagttccgc cgtggcaata gggaggggga
     3781 aagcgaaagt cccggaaagg agctgacagg tggtggcaat gccccaacca gtgggggttg
     3841 cgtcagcaaa cacagtgcac accacgccac gttgcctgac aacgggccac aactcctcat
     3901 aaagagacag caaccaggat ttatacaagg aggagaaaat gaaagccata cgggaagcaa
     3961 tagcatgata caaaggcatt aaagcagcgt atccacatag cgtaaaagga gcaacatagt
     4021 taagaatacc agtcaatctt tcacaaattt tgtaatccag aggttgatta tcgataagct
     4081 tgatatcgaa ttcgaagttc ctattcttca aaaggtatag gaacttctct ttgcaccatt
     4141 ctaaagaata acagtgataa tttctgggtt aaggcaagaa gttcctattc ttcaaatagt
     4201 ataggaactt cgagctcgcc accatggact atggcggcgc tttgtctgcc gtcggacgcg
     4261 aacttttgtt cgttactaat cctgtggtgg tgaacgggtc cgtcctggtc cctgaggatc
     4321 aatgttactg tgccggatgg attgaatctc gcggcacgaa cggcgctcag accgcgtcaa
     4381 atgtcctgca gtggcttgca gcaggattca gcattttgct gctgatgttc tatgcctacc
     4441 aaacctggaa atctacatgc ggctgggagg agatctatgt gtgcgccatt gaaatggtta
     4501 aggtgattct cgagttcttt tttgagttta agaatccctc tatgctctac cttgccacag
     4561 gacaccgggt gcagtggctg cgctatgcag agtggctgct cacttgtcct gtcatcctta
     4621 tccgcctgag caacctcacc ggtaagtgtc ggggttgcta gcataacttc gtatagtata
     4681 ccttatacga agttattctt tgcaccattc taaagaataa cagtgataat ttctgggtta
     4741 aggcaaataa cttcgtatag gatactttat acgaagttat ggtacccctc ctgtactcac
     4801 cttgccggtg gtgcagatga acttcagggt cagcttgccg taggtggcat cgccctcgcc
     4861 ctcgccggac acgctgaact tgtggccgtt tacgtcgccg tccagctcga ccaggatggg
     4921 caccaccccg gtgaacagct cctcgccctt gctcacggcg gccgctggca cggctccggc
     4981 ctcggcttcg tcttcgacga gagtctcgac ctcgatctcc gttccgccga tgttcagttt
     5041 ggtggttttg cggatatctc cgtgaatcaa tatgtgctcg tggatcagga cgcgcaggta
     5101 gtgtcccaac aacccccaac aatttttact catcagatca ataatcgtgt gacctacggt
     5161 ggagccatag acgctcagga cgccaaaacc ttcgggcccc aaaatgaaga gaattgggaa
     5221 cataccccag ctcacgaaaa acagccatgc catgccggtc acgacctggc ggcaccgacc
     5281 ctttggcaca gtatgataac cctcgatata tgctttggcg gcgtgaaaaa atgtgttcgc
     5341 gccatagcac aatccaagac aaaagaagat gactttaaca tagccggttg ccatggcgct
     5401 ggtagccccc cacacgatag tcccgatgtc tgagacaagg agtcccatgg ttctcctgct
     5461 gtagtcgttg ctcaggccta aacacatgag aaatagagtc ataagcacat gggtcacata
     5521 caggagatat gtatataaca ttaatacaat ttataaaaaa aagggggggc acaactagta
     5581 taacttcgta taaggtatac tatacgaagt tattaagagg tttcatattg ctaatagcag
     5641 ctacaatcca gctaccattc tgcataactt cgtataaagt atcctatacg aagttatgtc
     5701 gactggagag tggccagccc ttctcatgtt cagagaacat ggttaactgg ttaagtcatg
     5761 tcgtcccaca gctgcccgtg ccctggccca ccctcgtgac caccttcggc tacggcctgc
     5821 agtgcttcgc ccgctacccc gaccacatga agcagcacga cttcttcaag tccgccatgc
     5881 ccgaaggcta cgtccaggag cgcaccatct tcttcaagga cgacggcaac tacaagaccc
     5941 gcgccgaggt gaagttcgag ggcgacaccc tggtgaaccg catcgagctg aagggcatcg
     6001 acttcaagga ggacggcaac atcctggggc acaagctgga gtacaactac aacagccaca
     6061 acgtctatat catggccgac aagcagaaga acggcatcaa ggtgaacttc aagatccgcc
     6121 acaacatcga ggacggcagc gtgcagctcg ccgaccacta ccagcagaac acccccatcg
     6181 gcgacggccc cgtgctgctg cccgacaacc actacctgag ctaccagtcc gccctgagca
     6241 aagaccccaa cgagaagcgc gatcacatgg tcctgctgga gttcgtgacc gccgccggga
     6301 tcactctcgg catggacgag ctgtacaagt aaggcgcgcc gaagttccta taccttttga
     6361 agaataggaa cttctaagag gttttagatt gctaatagca gctacaatcc agctactact
     6421 ctgcgaagtt cctatactat ttgaagaata ggaacttcgg atccttctcg actgcgctct
     6481 caggcacgac acgactcctc cgctgcccac cgcagactga ggcagcgctg agtcgccggc
     6541 gccgcagcgc agatggtcgc gcccgtgccc ccctatctcg cgcctcgcgt ggtgcggtcc
     6601 ggctgggccg gcggcggcgc ggacgcgacc aaggtggccg ggaaggggag tttgcggggg
     6661 accggcgagt gacgtcagcg cgccttcagt gctgaggcgg cggtggcgcg cgccgccagg
     6721 cgggggcgaa ggcactgtcc gcggtgctga agctggcagt gcgcacgcgc ctcgccgcat
     6781 cctgtttccc ctccccctct ctgatagggg atgcgcaatt tggggaatgg gggttgggtg
     6841 cttgtccagt gggtcggggt cggtcgtcag gtaggcaccc ccaccccgcc tcatcctg
//
