LOCUS Exported 6897 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION AAV vector with hSynapsin promoter, Cre-OFF mCAR (for efficient CAV-2 infection) and Cre-ON ChETA (for optogenetic activation). ACCESSION . VERSION . KEYWORDS pAAV-hSyn-DIO {mCAR}off-{ChETA}on-WPRE SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6897) AUTHORS Li SJ, Vaughan A, Sturgill JF, Kepecs A TITLE A Viral Receptor Complementation Strategy to Overcome CAV-2 Tropism for Efficient Retrograde Targeting of Neurons. JOURNAL Neuron. 2018 Jun 6;98(5):905-917.e5. doi: 10.1016/j.neuron.2018.05.028. PUBMED 29879392 REFERENCE 2 (bases 1 to 6897) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..6897 /organism="synthetic DNA construct" /mol_type="other DNA" repeat_region 2..142 /label=AAV2 ITR /note="inverted terminal repeat of adeno-associated virus serotype 2" repeat_region 13..142 /label=AAV2 ITR promoter 187..634 /label=hSyn promoter /note="human synapsin I promoter; confers neuron-specific expression (Kugler et al., 2003)" protein_bind 672..705 /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)." protein_bind 756..789 /label=loxP /bound_moiety="Cre recombinase" /note="Cre-mediated recombination occurs in the 8-bp core sequence (GCATACAT)." regulatory 795..804 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 1857..1886 /codon_start=1 /product="Myc (human c-Myc proto-oncogene) epitope tag" /label=Myc /translation="EQKLISEEDL" CDS complement(1901..1927) /codon_start=1 /product="HA (human influenza hemagglutinin) epitope tag" /label=HA /translation="YPYDVPDYA" CDS complement(1946..2872) /codon_start=1 /product="H134R mutant of an N-terminal fragment of channelrhodopsin-2 from the alga Chlamydomonas reinhardtii (Nagel et al., 2005)" /label=ChR2(H134R) /note="blue light-activated cation channel" /translation="MDYGGALSAVGRELLFVTNPVVVNGSVLVPEDQCYCAGWIESRGT NGAQTASNVLQWLAAGFSILLLMFYAYQTWKSTCGWEEIYVCAIEMVKVILEFFFEFKN PSMLYLATGHRVQWLRYATWLLTCPVILIRLSNLTGLSNDYSRRTMGLLVSDIGTIVWG ATSAMATGYVKVIFFCLGLCYGANTFFHAAKAYIEGYHTVPKGRCRQVVTGMAWLFFVS WGMFPILFILGPEGFGVLSVYGSTVGHTIIDLMSKNCWGLLGHYLRVLIHEHILIHGDI RKTTKLNIGGTEIEVETLVEDEAEAGAV" regulatory 2869..2878 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" protein_bind complement(2883..2916) /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)." protein_bind complement(2967..3000) /label=loxP /bound_moiety="Cre recombinase" /note="Cre-mediated recombination occurs in the 8-bp core sequence (GCATACAT)." misc_feature 3025..3613 /label=WPRE /note="woodchuck hepatitis virus posttranscriptional regulatory element" primer_bind complement(3078..3098) /label=WPRE-R /note="WPRE, reverse primer" CDS complement(3496..3507) /codon_start=1 /product="Factor Xa recognition and cleavage site" /label=Factor Xa site /translation="IEGR" polyA_signal 3645..4121 /label=hGH poly(A) signal /note="human growth hormone polyadenylation signal" primer_bind complement(3844..3863) /label=hGH-PA-R /note="Human growth hormone terminator, reverse primer" repeat_region 4161..4301 /label=AAV2 ITR /note="inverted terminal repeat of adeno-associated virus serotype 2" repeat_region 4161..4290 /label=AAV2 ITR rep_origin 4376..4831 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(4463..4482) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 4673..4694 /label=F1ori-F /note="F1 origin, forward primer" primer_bind complement(4848..4867) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind 4967..4989 /label=pGEX 3' /note="pGEX vectors, reverse primer" primer_bind complement(5027..5045) /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" promoter 5113..5217 /gene="bla" /label=AmpR promoter CDS 5218..6078 /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(5436..5455) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 6249..6837 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 6738..6757 /label=pBR322ori-F /note="pBR322 origin, forward primer" ORIGIN 1 tcctgcaggc agctgcgcgc tcgctcgctc actgaggccg cccgggcaaa gcccgggcgt 61 cgggcgacct ttggtcgccc ggcctcagtg agcgagcgag cgcgcagaga gggagtggcc 121 aactccatca ctaggggttc ctgcggccgc acgcgtgtgt ctagactgca gagggccctg 181 cgtatgagtg caagtgggtt ttaggaccag gatgaggcgg ggtgggggtg cctacctgac 241 gaccgacccc gacccactgg acaagcaccc aacccccatt ccccaaattg cgcatcccct 301 atcagagagg gggaggggaa acaggatgcg gcgaggcgcg tgcgcactgc cagcttcagc 361 accgcggaca gtgccttcgc ccccgcctgg cggcgcgcgc caccgccgcc tcagcactga 421 aggcgcgctg acgtcactcg ccggtccccc gcaaactccc cttcccggcc accttggtcg 481 cgtccgcgcc gccgccggcc cagccggacc gcaccacgcg aggcgcgaga taggggggca 541 cgggcgcgac catctgcgct gcggcgccgg cgactcagcg ctgcctcagt ctgcggtggg 601 cagcggagga gtcgtgtcgt gcctgagagc gcagtcgaga aggtaccgga tcctctagag 661 tcgactccgg aataacttcg tataggatac tttatacgaa gttatgcaga atggtagctg 721 gattgtagct gctattagca atatgaaacc tcttaataac ttcgtatagc atacattata 781 cgaagttatg gcgcgccacc atggctcgcc tactgtgctt cgtgctcttg tgcgggatcg 841 cggatttcac cagtggtttg agcatcacta cacccgaaca gaggatcgaa aaagccaaag 901 gggaaactgc gtatctacca tgcaagttta ctctcagtcc cgaagaccag ggaccactgg 961 acattgaatg gctgatatcc ccgtctgata accagatagt ggatcaagtg atcattttgt 1021 attctggaga caaaatttat gataactact atccggatct gaaaggacgg gtacatttta 1081 cgagtaacga tgtcaagtct ggcgacgcat ctataaatgt gaccaacctg cagctgtcgg 1141 acattggcac ttaccagtgc aaagtgaaga aagcccctgg ggttgcaaat aagaaattcc 1201 tgctgaccgt tcttgttaag ccttcaggta caagatgctt cgtggatgga tcggaagaga 1261 ttggaaatga cttcaagcta aaatgtgaac ccaaggaagg ctcccttcca ctacagtttg 1321 aatggcagaa actgtcggac tcccagacaa tgcctacgcc atggctggca gaaatgacgt 1381 caccagttat atctgtgaag aacgccagtt ctgagtattc tgggacatac agctgcacgg 1441 ttcaaaacag agtgggctct gaccagtgta tgctgcgact agacgttgtc ccaccctcca 1501 accgagccgg aacgatcgcg ggcgccgtca tagggacgct gctggccctt gtgctcatcg 1561 gggccatcct cttctgctgt cacaggaaac gcagagaaga gaagtacgag aaggaagttc 1621 atcatgatat cagggaagat gtgcctcctc caaagagtcg gacatccact gccaggagct 1681 atattggcag caaccattcc tccctgggat ccatgtcccc ctctaacatg gaggggtatt 1741 ccaagacgca gtataaccaa gtccccagtg aagactttga acgtgcgcct cagagcccga 1801 ctctggcacc cgctaagttc aagtacgctt acaagaccga tggcattaca gtggtagagc 1861 agaagctgat cagcgaggag gacctgtaat taattaatta ggcgtagtcg ggcacgtcgt 1921 aggggtacat ggtggcggcc gctggcacgg ctccggcctc ggcttcgtct tcgacgagag 1981 tctcgacctc gatctccgtt ccgccgatgt tcagtttggt ggttttgcgg atatctccgt 2041 gaatcaatat gtgctcgtgg atcaggacgc gcaggtagtg tcccaacaac ccccaacaat 2101 ttttactcat cagatcaata atcgtgtgac ctacggtgga gccatagacg ctcaggacgc 2161 caaaaccttc gggccccaaa atgaagagaa ttgggaacat accccagctc acgaaaaaca 2221 gccatgccat gccggtcacg acctggcggc accgaccctt tggcacagta tgataaccct 2281 cgatatatgc tttggcggcg tgaaaaaatg tgttcgcgcc atagcacaat ccaagacaaa 2341 agaagatgac tttaacatag ccggttgcca tggcgctggt agccccccac acgatagtcc 2401 cgatgtctga gacaaggagt cccatggttc tcctgctgta gtcgttgctc aggccggtga 2461 ggttgctcag gcggataagg atgacaggac aagtgagcag ccaggttgca tagcgcagcc 2521 actgcacccg gtgtcctgtg gcaaggtaga gcatagaggg attcttaaac tcaaaaaaga 2581 actcgagaat caccttaacc atttcaatgg cgcacacata gatctcctcc cagccgcatg 2641 tagatttcca ggtttggtag gcatagaaca tcagcagcaa aatgctgaat cctgctgcaa 2701 gccactgcag gacatttgac gcggtctgag cgccgttcgt gccgcgagat tcaatccatc 2761 cggcacagta acattgatcc tcagggacca ggacggaccc gttcaccacc acaggattag 2821 taacgaacaa aagttcgcgt ccgacggcag acaaagcgcc gccatagtcc atggtggcta 2881 gcataacttc gtataaagta tcctatacga agttatttgc cttaacccag aaattatcac 2941 tgttattctt tagaatggtg caaagaataa cttcgtataa tgtatgctat acgaagttat 3001 gaattcgata tcaagcttat cgataatcaa cctctggatt acaaaatttg tgaaagattg 3061 actggtattc ttaactatgt tgctcctttt acgctatgtg gatacgctgc tttaatgcct 3121 ttgtatcatg ctattgcttc ccgtatggct ttcattttct cctccttgta taaatcctgg 3181 ttgctgtctc tttatgagga gttgtggccc gttgtcaggc aacgtggcgt ggtgtgcact 3241 gtgtttgctg acgcaacccc cactggttgg ggcattgcca ccacctgtca gctcctttcc 3301 gggactttcg ctttccccct ccctattgcc acggcggaac tcatcgccgc ctgccttgcc 3361 cgctgctgga caggggctcg gctgttgggc actgacaatt ccgtggtgtt gtcggggaaa 3421 tcatcgtcct ttccttggct gctcgcctat gttgccacct ggattctgcg cgggacgtcc 3481 ttctgctacg tcccttcggc cctcaatcca gcggaccttc cttcccgcgg cctgctgccg 3541 gctctgcggc ctcttccgcg tcttcgcctt cgccctcaga cgagtcggat ctccctttgg 3601 gccgcctccc cgcatcgata ccgagcgctg ctcgagagat ctacgggtgg catccctgtg 3661 acccctcccc agtgcctctc ctggccctgg aagttgccac tccagtgccc accagccttg 3721 tcctaataaa attaagttgc atcattttgt ctgactaggt gtccttctat aatattatgg 3781 ggtggagggg ggtggtatgg agcaaggggc aagttgggaa gacaacctgt agggcctgcg 3841 gggtctattg ggaaccaagc tggagtgcag tggcacaatc ttggctcact gcaatctccg 3901 cctcctgggt tcaagcgatt ctcctgcctc agcctcccga gttgttggga ttccaggcat 3961 gcatgaccag gctcagctaa tttttgtttt tttggtagag acggggtttc accatattgg 4021 ccaggctggt ctccaactcc taatctcagg tgatctaccc accttggcct cccaaattgc 4081 tgggattaca ggcgtgaacc actgctccct tccctgtcct tctgattttg taggtaacca 4141 cgtgcggacc gagcggccgc aggaacccct agtgatggag ttggccactc cctctctgcg 4201 cgctcgctcg ctcactgagg ccgggcgacc aaaggtcgcc cgacgcccgg gctttgcccg 4261 ggcggcctca gtgagcgagc gagcgcgcag ctgcctgcag gggcgcctga tgcggtattt 4321 tctccttacg catctgtgcg gtatttcaca ccgcatacgt caaagcaacc atagtacgcg 4381 ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca 4441 cttgccagcg ccctagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc 4501 gccggctttc cccgtcaagc tctaaatcgg gggctccctt tagggttccg atttagtgct 4561 ttacggcacc tcgaccccaa aaaacttgat ttgggtgatg gttcacgtag tgggccatcg 4621 ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa tagtggactc 4681 ttgttccaaa ctggaacaac actcaaccct atctcgggct attcttttga tttataaggg 4741 attttgccga tttcggccta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg 4801 aattttaaca aaatattaac gtttacaatt ttatggtgca ctctcagtac aatctgctct 4861 gatgccgcat agttaagcca gccccgacac ccgccaacac ccgctgacgc gccctgacgg 4921 gcttgtctgc tcccggcatc cgcttacaga caagctgtga ccgtctccgg gagctgcatg 4981 tgtcagaggt tttcaccgtc atcaccgaaa cgcgcgagac gaaagggcct cgtgatacgc 5041 ctatttttat aggttaatgt catgataata atggtttctt agacgtcagg tggcactttt 5101 cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat 5161 ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtatg 5221 agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt 5281 tttgctcacc cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcacga 5341 gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa 5401 gaacgttttc caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt 5461 attgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt 5521 gagtactcac cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc 5581 agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac aacgatcgga 5641 ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat 5701 cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct 5761 gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc 5821 cggcaacaat taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg 5881 gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc 5941 ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg 6001 acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca 6061 ctgattaagc attggtaact gtcagaccaa gtttactcat atatacttta gattgattta 6121 aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc 6181 aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa 6241 ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca 6301 ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta 6361 actggcttca gcagagcgca gataccaaat actgtccttc tagtgtagcc gtagttaggc 6421 caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca 6481 gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta 6541 ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag 6601 cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt 6661 cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc 6721 acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac 6781 ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac 6841 gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc tcacatg //