LOCUS Exported 6909 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Cre-on/Flp-off ChR2-EYFP under the Synapsin promoter. ACCESSION . VERSION . KEYWORDS pAAV-hSyn Con/Foff hChR2(H134R)-EYFP 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 May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..6909 /organism="synthetic DNA construct" /mol_type="other DNA" 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)" regulatory 773..782 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" protein_bind 1218..1251 /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 1302..1335 /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 1448..1469 /label=EGFP-N /note="EGFP, reverse primer" protein_bind complement(2135..2168) /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(2219..2252) /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(2474..2493) /label=EXFP-R /note="For distinguishing EGFP variants, reverse primer" CDS 2633..2884 /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 2821..2842 /label=EGFP-C /note="EGFP, forward primer" 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 5229..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 ccgccaccat 781 ggactatggc ggcgctttgt ctgccgtcgg acgcgaactt ttgttcgtta ctaatcctgt 841 ggtggtgaac gggtccgtcc tggtccctga ggatcaatgt tactgtgccg gatggattga 901 atctcgcggc acgaacggcg ctcagaccgc gtcaaatgtc ctgcagtggc ttgcagcagg 961 attcagcatt ttgctgctga tgttctatgc ctaccaaacc tggaaatcta catgcggctg 1021 ggaggagatc tatgtgtgcg ccattgaaat ggttaaggtg attctcgagt tcttttttga 1081 gtttaagaat ccctctatgc tctaccttgc cacaggacac cgggtgcagt ggctgcgcta 1141 tgcagagtgg ctgctcactt gtcctgtcat ccttatccgc ctgagcaacc tcaccggtaa 1201 gtgtcggggt tgctagcata acttcgtata gtatacctta tacgaagtta ttctttgcac 1261 cattctaaag aataacagtg ataatttctg ggttaaggca aataacttcg tataggatac 1321 tttatacgaa gttatggtac ccctcctgta ctcaccttgc cggtggtgca gatgaacttc 1381 agggtcagct tgccgtaggt ggcatcgccc tcgccctcgc cggacacgct gaacttgtgg 1441 ccgtttacgt cgccgtccag ctcgaccagg atgggcacca ccccggtgaa cagctcctcg 1501 cccttgctca cggcggccgc tggcacggct ccggcctcgg cttcgtcttc gacgagagtc 1561 tcgacctcga tctccgttcc gccgatgttc agtttggtgg ttttgcggat atctccgtga 1621 atcaatatgt gctcgtggat caggacgcgc aggtagtgtc ccaacaaccc ccaacaattt 1681 ttactcatca gatcaataat cgtgtgacct acggtggagc catagacgct caggacgcca 1741 aaaccttcgg gccccaaaat gaagagaatt gggaacatac cccagctcac gaaaaacagc 1801 catgccatgc cggtcacgac ctggcggcac cgaccctttg gcacagtatg ataaccctcg 1861 atatatgctt tggcggcgtg aaaaaatgtg ttcgcgccat agcacaatcc aagacaaaag 1921 aagatgactt taacatagcc ggttgccatg gcgctggtag ccccccacac gatagtcccg 1981 atgtctgaga caaggagtcc catggttctc ctgctgtagt cgttgctcag gcctaaacac 2041 atgagaaata gagtcataag cacatgggtc acatacagga gatatgtata taacattaat 2101 acaatttata aaaaaaaggg ggggcacaac tagtataact tcgtataagg tatactatac 2161 gaagttatta agaggtttca tattgctaat agcagctaca atccagctac cattctgcat 2221 aacttcgtat aaagtatcct atacgaagtt atgtcgactg gagagtggcc agcccttctc 2281 atgttcagag aacatggtta actggttaag tcatgtcgtc ccacagctgc ccgtgccctg 2341 gcccaccctc gtgaccacct tcggctacgg cctgcagtgc ttcgcccgct accccgacca 2401 catgaagcag cacgacttct tcaagtccgc catgcccgaa ggctacgtcc aggagcgcac 2461 catcttcttc aaggacgacg gcaactacaa gacccgcgcc gaggtgaagt tcgagggcga 2521 caccctggtg aaccgcatcg agctgaaggg catcgacttc aaggaggacg gcaacatcct 2581 ggggcacaag ctggagtaca actacaacag ccacaacgtc tatatcatgg ccgacaagca 2641 gaagaacggc atcaaggtga acttcaagat ccgccacaac atcgaggacg gcagcgtgca 2701 gctcgccgac cactaccagc agaacacccc catcggcgac ggccccgtgc tgctgcccga 2761 caaccactac ctgagctacc agtccgccct gagcaaagac cccaacgaga agcgcgatca 2821 catggtcctg ctggagttcg tgaccgccgc cgggatcact ctcggcatgg acgagctgta 2881 caagtaagag 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 //