LOCUS Exported 6423 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Channelrhodopsin localized to neuronal soma via a Kv2.1 targeting motif. ACCESSION . VERSION . KEYWORDS AAV-syn-ChR2-EYFP-Kv SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6423) AUTHORS Baker CA, Elyada YM, Parra A, Bolton MM TITLE Cellular resolution circuit mapping with temporal-focused excitation of soma-targeted channelrhodopsin. JOURNAL Elife. 2016 Aug 15;5. pii: e14193. doi: 10.7554/eLife.14193. PUBMED 27525487 REFERENCE 2 (bases 1 to 6423) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..6423 /organism="synthetic DNA construct" /mol_type="other DNA" regulatory 229..238 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 235..1161 /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 PSMLYLATGHRVQWLRYAEWLLTCPVILIRLSNLTGLSNDYSRRTMGLLVSDIGTIVWG ATSAMATGYVKVIFFCLGLCYGANTFFHAAKAYIEGYHTVPKGRCRQVVTGMAWLFFVS WGMFPILFILGPEGFGVLSVYGSTVGHTIIDLMSKNCWGLLGHYLRVLIHEHILIHGDI RKTTKLNIGGTEIEVETLVEDEAEAGAV" regulatory 1171..1180 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 1177..1893 /codon_start=1 /product="enhanced YFP" /label=EYFP /note="mammalian codon-optimized" /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KFICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL EFVTAAGITLGMDELYK" primer_bind complement(1222..1243) /label=EGFP-N /note="EGFP, reverse primer" primer_bind complement(1483..1502) /label=EXFP-R /note="For distinguishing EGFP variants, reverse primer" primer_bind 1830..1851 /label=EGFP-C /note="EGFP, forward primer" misc_feature 2119..2707 /label=WPRE /note="woodchuck hepatitis virus posttranscriptional regulatory element" primer_bind complement(2172..2192) /label=WPRE-R /note="WPRE, reverse primer" CDS complement(2590..2601) /codon_start=1 /product="Factor Xa recognition and cleavage site" /label=Factor Xa site /translation="IEGR" polyA_signal 2739..3215 /label=hGH poly(A) signal /note="human growth hormone polyadenylation signal" primer_bind complement(2938..2957) /label=hGH-PA-R /note="Human growth hormone terminator, reverse primer" repeat_region 3255..3395 /label=AAV2 ITR /note="inverted terminal repeat of adeno-associated virus serotype 2" repeat_region 3255..3384 /label=AAV2 ITR rep_origin 3470..3925 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(3557..3576) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 3767..3788 /label=F1ori-F /note="F1 origin, forward primer" primer_bind complement(3942..3961) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind 4061..4083 /label=pGEX 3' /note="pGEX vectors, reverse primer" primer_bind complement(4121..4139) /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" promoter 4207..4311 /gene="bla" /label=AmpR promoter CDS 4312..5172 /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(4530..4549) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 5343..5931 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 5832..5851 /label=pBR322ori-F /note="pBR322 origin, forward primer" repeat_region 5993..6122 /label=AAV2 ITR (alternate) /note="Functional equivalent of wild-type AAV2 ITR" promoter 6167..191 /label=hSyn promoter /note="human synapsin I promoter; confers neuron-specific expression (Kugler et al., 2003)" ORIGIN 1 gtcccccgca aactcccctt cccggccacc ttggtcgcgt ccgcgccgcc gccggcccag 61 ccggaccgca ccacgcgagg cgcgagatag gggggcacgg gcgcgaccat ctgcgctgcg 121 gcgccggcga ctcagcgctg cctcagtctg cggtgggcag cggaggagtc gtgtcgtgcc 181 tgagagcgca gtcgagaagg taccggatcc tctagagtcg actccggagc caccatggac 241 tatggcggcg ctttgtctgc cgtcggacgc gaacttttgt tcgttactaa tcctgtggtg 301 gtgaacgggt ccgtcctggt ccctgaggat caatgttact gtgccggatg gattgaatct 361 cgcggcacga acggcgctca gaccgcgtca aatgtcctgc agtggcttgc agcaggattc 421 agcattttgc tgctgatgtt ctatgcctac caaacctgga aatctacatg cggctgggag 481 gagatctatg tgtgcgccat tgaaatggtt aaggtgattc tcgagttctt ttttgagttt 541 aagaatccct ctatgctcta ccttgccaca ggacaccggg tgcagtggct gcgctatgca 601 gagtggctgc tcacttgtcc tgtcatcctt atccgcctga gcaacctcac cggcctgagc 661 aacgactaca gcaggagaac catgggactc cttgtctcag acatcgggac tatcgtgtgg 721 ggggctacca gcgccatggc aaccggctat gttaaagtca tcttcttttg tcttggattg 781 tgctatggcg cgaacacatt ttttcacgcc gccaaagcat atatcgaggg ttatcatact 841 gtgccaaagg gtcggtgccg ccaggtcgtg accggcatgg catggctgtt tttcgtgagc 901 tggggtatgt tcccaattct cttcattttg gggcccgaag gttttggcgt cctgagcgtc 961 tatggctcca ccgtaggtca cacgattatt gatctgatga gtaaaaattg ttgggggttg 1021 ttgggacact acctgcgcgt cctgatccac gagcacatat tgattcacgg agatatccgc 1081 aaaaccacca aactgaacat cggcggaacg gagatcgagg tcgagactct cgtcgaagac 1141 gaagccgagg ccggagccgt gccagcggcc gccaccatgg tgagcaaggg cgaggagctg 1201 ttcaccgggg tggtgcccat cctggtcgag ctggacggcg acgtaaacgg ccacaagttc 1261 agcgtgtccg gcgagggcga gggcgatgcc acctacggca agctgaccct gaagttcatc 1321 tgcaccaccg gcaagctgcc cgtgccctgg cccaccctcg tgaccacctt cggctacggc 1381 ctgcagtgct tcgcccgcta ccccgaccac atgaagcagc acgacttctt caagtccgcc 1441 atgcccgaag gctacgtcca ggagcgcacc atcttcttca aggacgacgg caactacaag 1501 acccgcgccg aggtgaagtt cgagggcgac accctggtga accgcatcga gctgaagggc 1561 atcgacttca aggaggacgg caacatcctg gggcacaagc tggagtacaa ctacaacagc 1621 cacaacgtct atatcatggc cgacaagcag aagaacggca tcaaggtgaa cttcaagatc 1681 cgccacaaca tcgaggacgg cagcgtgcag ctcgccgacc actaccagca gaacaccccc 1741 atcggcgacg gccccgtgct gctgcccgac aaccactacc tgagctacca gtccgccctg 1801 agcaaagacc ccaacgagaa gcgcgatcac atggtcctgc tggagttcgt gaccgccgcc 1861 gggatcactc tcggcatgga cgagctgtac aagcagtccc agcctattct gaacaccaaa 1921 gagatggccc cccagagtaa accccctgag gagctggaaa tgtcctcaat gccctcccct 1981 gtggcacctc ttcctgcaag aacagagggc gtgatcgaca tgaggtccat gagttctatt 2041 gactctttta tctcatgcgc caccgatttt ccagaggcta cgcggtttaa gtaagaattc 2101 gatatcaagc ttatcgataa tcaacctctg gattacaaaa tttgtgaaag attgactggt 2161 attcttaact atgttgctcc ttttacgcta tgtggatacg ctgctttaat gcctttgtat 2221 catgctattg cttcccgtat ggctttcatt ttctcctcct tgtataaatc ctggttgctg 2281 tctctttatg aggagttgtg gcccgttgtc aggcaacgtg gcgtggtgtg cactgtgttt 2341 gctgacgcaa cccccactgg ttggggcatt gccaccacct gtcagctcct ttccgggact 2401 ttcgctttcc ccctccctat tgccacggcg gaactcatcg ccgcctgcct tgcccgctgc 2461 tggacagggg ctcggctgtt gggcactgac aattccgtgg tgttgtcggg gaaatcatcg 2521 tcctttcctt ggctgctcgc ctatgttgcc acctggattc tgcgcgggac gtccttctgc 2581 tacgtccctt cggccctcaa tccagcggac cttccttccc gcggcctgct gccggctctg 2641 cggcctcttc cgcgtcttcg ccttcgccct cagacgagtc ggatctccct ttgggccgcc 2701 tccccgcatc gataccgagc gctgctcgag agatctacgg gtggcatccc tgtgacccct 2761 ccccagtgcc tctcctggcc ctggaagttg ccactccagt gcccaccagc cttgtcctaa 2821 taaaattaag ttgcatcatt ttgtctgact aggtgtcctt ctataatatt atggggtgga 2881 ggggggtggt atggagcaag gggcaagttg ggaagacaac ctgtagggcc tgcggggtct 2941 attgggaacc aagctggagt gcagtggcac aatcttggct cactgcaatc tccgcctcct 3001 gggttcaagc gattctcctg cctcagcctc ccgagttgtt gggattccag gcatgcatga 3061 ccaggctcag ctaatttttg tttttttggt agagacgggg tttcaccata ttggccaggc 3121 tggtctccaa ctcctaatct caggtgatct acccaccttg gcctcccaaa ttgctgggat 3181 tacaggcgtg aaccactgct cccttccctg tccttctgat tttgtaggta accacgtgcg 3241 gaccgagcgg ccgcaggaac ccctagtgat ggagttggcc actccctctc tgcgcgctcg 3301 ctcgctcact gaggccgggc gaccaaaggt cgcccgacgc ccgggctttg cccgggcggc 3361 ctcagtgagc gagcgagcgc gcagctgcct gcaggggcgc ctgatgcggt attttctcct 3421 tacgcatctg tgcggtattt cacaccgcat acgtcaaagc aaccatagta cgcgccctgt 3481 agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc 3541 agcgccttag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc 3601 tttccccgtc aagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg 3661 cacctcgacc ccaaaaaact tgatttgggt gatggttcac gtagtgggcc atcgccctga 3721 tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc 3781 caaactggaa caacactcaa ctctatctcg ggctattctt ttgatttata agggattttg 3841 ccgatttcgg tctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaatttt 3901 aacaaaatat taacgtttac aattttatgg tgcactctca gtacaatctg ctctgatgcc 3961 gcatagttaa gccagccccg acacccgcca acacccgctg acgcgccctg acgggcttgt 4021 ctgctcccgg catccgctta cagacaagct gtgaccgtct ccgggagctg catgtgtcag 4081 aggttttcac cgtcatcacc gaaacgcgcg agacgaaagg gcctcgtgat acgcctattt 4141 ttataggtta atgtcatgat aataatggtt tcttagacgt caggtggcac ttttcgggga 4201 aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc 4261 atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag tatgagtatt 4321 caacatttcc gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct 4381 cacccagaaa cgctggtgaa agtaaaagat gctgaagatc agttgggtgc acgagtgggt 4441 tacatcgaac tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgt 4501 tttccaatga tgagcacttt taaagttctg ctatgtggcg cggtattatc ccgtattgac 4561 gccgggcaag agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtac 4621 tcaccagtca cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct 4681 gccataacca tgagtgataa cactgcggcc aacttacttc tgacaacgat cggaggaccg 4741 aaggagctaa ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg 4801 gaaccggagc tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgtagca 4861 atggcaacaa cgttgcgcaa actattaact ggcgaactac ttactctagc ttcccggcaa 4921 caattaatag actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt 4981 ccggctggct ggtttattgc tgataaatct ggagccggtg agcgtgggtc tcgcggtatc 5041 attgcagcac tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg 5101 agtcaggcaa ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt 5161 aagcattggt aactgtcaga ccaagtttac tcatatatac tttagattga tttaaaactt 5221 catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc 5281 ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct 5341 tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta 5401 ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc 5461 ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt aggccaccac 5521 ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct 5581 gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat 5641 aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg 5701 acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa 5761 gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg 5821 gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga 5881 cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc 5941 aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat gtcctgcagg 6001 cagctgcgcg ctcgctcgct cactgaggcc gcccgggcgt cgggcgacct ttggtcgccc 6061 ggcctcagtg agcgagcgag cgcgcagaga gggagtggcc aactccatca ctaggggttc 6121 ctgcggccgc acgcgtgtgt ctagactgca gagggccctg cgtatgagtg caagtgggtt 6181 ttaggaccag gatgaggcgg ggtgggggtg cctacctgac gaccgacccc gacccactgg 6241 acaagcaccc aacccccatt ccccaaattg cgcatcccct atcagagagg gggaggggaa 6301 acaggatgcg gcgaggcgcg tgcgcactgc cagcttcagc accgcggaca gtgccttcgc 6361 ccccgcctgg cggcgcgcgc caccgccgcc tcagcactga aggcgcgctg acgtcactcg 6421 ccg //