LOCUS Exported 7035 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION AAV expression of CaMKII-driven stabilized step function opsin (SSFO) for optogenetic activation.. ACCESSION . VERSION . KEYWORDS pAAV-CaMKIIa-hChR2(C128S/D156A)-EYFP SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 7035) AUTHORS Yizhar O, Fenno LE, Prigge M, Schneider F, Davidson TJ, O'Shea DJ, Sohal VS, Goshen I, Finkelstein J, Paz JT, Stehfest K, Fudim R, Ramakrishnan C, Huguenard JR, Hegemann P, Deisseroth K TITLE Neocortical excitation/inhibition balance in information processing and social dysfunction. JOURNAL Nature. 2011 Jul 27;477(7363):171-8. doi: 10.1038/nature10360. PUBMED 21796121 REFERENCE 2 (bases 1 to 7035) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..7035 /organism="synthetic DNA construct" /mol_type="other DNA" repeat_region 49..178 /label=AAV2 ITR (alternate) /note="Functional equivalent of wild-type AAV2 ITR" promoter 197..1485 /label=CaMKII promoter /note="promoter for the mouse calcium/calmodulin-dependent kinase II alpha subunit" regulatory 1518..1527 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 1524..2450 /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 PSMLYLATGHRVQWLRYAEWLLTSPVILIHLSNLTGLSNDYSRRTMGLLVSAIGTIVWG ATSAMATGYVKVIFFCLGLCYGANTFFHAAKAYIEGYHTVPKGRCRQVVTGMAWLFFVS WGMFPILFILGPEGFGVLSVYGSTVGHTIIDLMSKNCWGLLGHYLRVLIHEHILIHGDI RKTTKLNIGGTEIEVETLVEDEAEAGAV" regulatory 2460..2469 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 2466..3185 /codon_start=1 /product="enhanced YFP" /label=EYFP /note="mammalian codon-optimized" /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KFICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL EFVTAAGITLGMDELYK" primer_bind complement(2511..2532) /label=EGFP-N /note="EGFP, reverse primer" primer_bind complement(2772..2791) /label=EXFP-R /note="For distinguishing EGFP variants, reverse primer" primer_bind 3119..3140 /label=EGFP-C /note="EGFP, forward primer" misc_feature 3210..3798 /label=WPRE /note="woodchuck hepatitis virus posttranscriptional regulatory element" primer_bind complement(3263..3283) /label=WPRE-R /note="WPRE, reverse primer" CDS complement(3681..3692) /codon_start=1 /product="Factor Xa recognition and cleavage site" /label=Factor Xa site /translation="IEGR" polyA_signal 3830..4306 /label=hGH poly(A) signal /note="human growth hormone polyadenylation signal" primer_bind complement(4029..4048) /label=hGH-PA-R /note="Human growth hormone terminator, reverse primer" repeat_region 4346..4486 /label=AAV2 ITR /note="inverted terminal repeat of adeno-associated virus serotype 2" repeat_region 4346..4475 /label=AAV2 ITR rep_origin 4561..5016 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(4648..4667) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 4858..4879 /label=F1ori-F /note="F1 origin, forward primer" primer_bind complement(5033..5052) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind 5152..5174 /label=pGEX 3' /note="pGEX vectors, reverse primer" primer_bind complement(5212..5230) /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" promoter 5298..5402 /gene="bla" /label=AmpR promoter CDS 5403..6263 /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(5621..5640) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 6434..7022 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 6923..6942 /label=pBR322ori-F /note="pBR322 origin, forward primer" ORIGIN 1 cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgtcc tgcaggcagc 61 tgcgcgctcg ctcgctcact gaggccgccc gggcgtcggg cgacctttgg tcgcccggcc 121 tcagtgagcg agcgagcgcg cagagaggga gtggccaact ccatcactag gggttcctgc 181 ggccgcacgc gtttaacatt atggccttag gtcacttcat ctccatgggg ttcttcttct 241 gattttctag aaaatgagat gggggtgcag agagcttcct cagtgacctg cccagggtca 301 catcagaaat gtcagagcta gaacttgaac tcagattact aatcttaaat tccatgcctt 361 gggggcatgc aagtacgata tacagaagga gtgaactcat tagggcagat gaccaatgag 421 tttaggaaag aagagtccag ggcagggtac atctacacca cccgcccagc cctgggtgag 481 tccagccacg ttcacctcat tatagttgcc tctctccagt cctaccttga cgggaagcac 541 aagcagaaac tgggacagga gccccaggag accaaatctt catggtccct ctgggaggat 601 gggtggggag agctgtggca gaggcctcag gaggggccct gctgctcagt ggtgacagat 661 aggggtgaga aagcagacag agtcattccg tcagcattct gggtctgttt ggtacttctt 721 ctcacgctaa ggtggcggtg tgatatgcac aatggctaaa aagcagggag agctggaaag 781 aaacaaggac agagacagag gccaagtcaa ccagaccaat tcccagagga agcaaagaaa 841 ccattacaga gactacaagg gggaagggaa ggagagatga attagcttcc cctgtaaacc 901 ttagaaccca gctgttgcca gggcaacggg gcaatacctg tctcttcaga ggagatgaag 961 ttgccagggt aactacatcc tgtctttctc aaggaccatc ccagaatgtg gcacccacta 1021 gccgttacca tagcaactgc ctctttgccc cacttaatcc catcccgtct gttaaaaggg 1081 ccctatagtt ggaggtgggg gaggtaggaa gagcgatgat cacttgtgga ctaagtttgt 1141 tcgcatcccc ttctccaacc ccctcagtac atcaccctgg gggaacaggg tccacttgct 1201 cctgggccca cacagtcctg cagtattgtg tatataaggc cagggcaaag aggagcaggt 1261 tttaaagtga aaggcaggca ggtgttgggg aggcagttac cggggcaacg ggaacagggc 1321 gtttcggagg tggttgccat ggggacctgg atgctgacga aggctcgcga ggctgtgagc 1381 agccacagtg ccctgctcag aagccccaag ctcgtcagtc aagccggttc tccgtttgca 1441 ctcaggagca cgggcaggcg agtggcccct agttctgggg gcagctctag agcggtaccg 1501 gatccccggg taccggtgcc accatggact atggcggcgc tttgtctgcc gtcggacgcg 1561 aacttttgtt cgttactaat cctgtggtgg tgaacgggtc cgtcctggtc cctgaggatc 1621 aatgttactg tgccggatgg attgaatctc gcggcacgaa cggcgctcag accgcgtcaa 1681 atgtcctgca gtggcttgca gcaggattca gcattttgct gctgatgttc tatgcctacc 1741 aaacctggaa atctacatgc ggctgggagg agatctatgt gtgcgccatt gaaatggtta 1801 aggtgattct cgagttcttt tttgagttta agaatccctc tatgctctac cttgccacag 1861 gacaccgggt gcagtggctg cgctatgcag agtggctgct cacttctcct gtcatcctta 1921 tccacctgag caacctcacc ggcctgagca acgactacag caggagaacc atgggactcc 1981 ttgtctcagc catcgggact atcgtgtggg gggctaccag cgccatggca accggctatg 2041 ttaaagtcat cttcttttgt cttggattgt gctatggcgc gaacacattt tttcacgccg 2101 ccaaagcata tatcgagggt tatcatactg tgccaaaggg tcggtgccgc caggtcgtga 2161 ccggcatggc atggctgttt ttcgtgagct ggggtatgtt cccaattctc ttcattttgg 2221 ggcccgaagg ttttggcgtc ctgagcgtct atggctccac cgtaggtcac acgattattg 2281 atctgatgag taaaaattgt tgggggttgt tgggacacta cctgcgcgtc ctgatccacg 2341 agcacatatt gattcacgga gatatccgca aaaccaccaa actgaacatc ggcggaacgg 2401 agatcgaggt cgagactctc gtcgaagacg aagccgaggc cggagccgtg ccagcggccg 2461 ccaccatggt gagcaagggc gaggagctgt tcaccggggt ggtgcccatc ctggtcgagc 2521 tggacggcga cgtaaacggc cacaagttca gcgtgtccgg cgagggcgag ggcgatgcca 2581 cctacggcaa gctgaccctg aagttcatct gcaccaccgg caagctgccc gtgccctggc 2641 ccaccctcgt gaccaccttc ggctacggcc tgcagtgctt cgcccgctac cccgaccaca 2701 tgaagcagca cgacttcttc aagtccgcca tgcccgaagg ctacgtccag gagcgcacca 2761 tcttcttcaa ggacgacggc aactacaaga cccgcgccga ggtgaagttc gagggcgaca 2821 ccctggtgaa ccgcatcgag ctgaagggca tcgacttcaa ggaggacggc aacatcctgg 2881 ggcacaagct ggagtacaac tacaacagcc acaacgtcta tatcatggcc gacaagcaga 2941 agaacggcat caaggtgaac ttcaagatcc gccacaacat cgaggacggc agcgtgcagc 3001 tcgccgacca ctaccagcag aacaccccca tcggcgacgg ccccgtgctg ctgcccgaca 3061 accactacct gagctaccag tccgccctga gcaaagaccc caacgagaag cgcgatcaca 3121 tggtcctgct ggagttcgtg accgccgccg ggatcactct cggcatggac gagctgtaca 3181 agtaagaatt cgatatcaag cttatcgata atcaacctct ggattacaaa atttgtgaaa 3241 gattgactgg tattcttaac tatgttgctc cttttacgct atgtggatac gctgctttaa 3301 tgcctttgta tcatgctatt gcttcccgta tggctttcat tttctcctcc ttgtataaat 3361 cctggttgct gtctctttat gaggagttgt ggcccgttgt caggcaacgt ggcgtggtgt 3421 gcactgtgtt tgctgacgca acccccactg gttggggcat tgccaccacc tgtcagctcc 3481 tttccggaac tttcgctttc cccctcccta ttgccacggc ggaactcatc gccgcctgcc 3541 ttgcccgctg ctggacaggg gctcggctgt tgggcactga caattccgtg gtgttgtcgg 3601 ggaaatcatc gtcctttcct tggctgctcg cctatgttgc cacctggatt ctgcgcggga 3661 cgtccttctg ctacgtccct tcggccctca atccagcgga ccttccttcc cgcggcctgc 3721 tgccggctct gcggcctctt ccgcgtcttc gccttcgccc tcagacgagt cggatctccc 3781 tttgggccgc ctccccgcat cgataccgag cgctgctcga gagatctacg ggtggcatcc 3841 ctgtgacccc tccccagtgc ctctcctggc cctggaagtt gccactccag tgcccaccag 3901 ccttgtccta ataaaattaa gttgcatcat tttgtctgac taggtgtcct tctataatat 3961 tatggggtgg aggggggtgg tatggagcaa ggggcaagtt gggaagacaa cctgtagggc 4021 ctgcggggtc tattgggaac caagctggag tgcagtggca caatcttggc tcactgcaat 4081 ctccgcctcc tgggttcaag cgattctcct gcctcagcct cccgagttgt tgggattcca 4141 ggcatgcatg accaggctca gctaattttt gtttttttgg tagagacggg gtttcaccat 4201 attggccagg ctggtctcca actcctaatc tcaggtgatc tacccacctt ggcctcccaa 4261 attgctggga ttacaggcgt gaaccactgc tcccttccct gtccttctga ttttgtaggt 4321 aaccacgtgc ggaccgagcg gccgcaggaa cccctagtga tggagttggc cactccctct 4381 ctgcgcgctc gctcgctcac tgaggccggg cgaccaaagg tcgcccgacg cccgggcttt 4441 gcccgggcgg cctcagtgag cgagcgagcg cgcagctgcc tgcaggggcg cctgatgcgg 4501 tattttctcc ttacgcatct gtgcggtatt tcacaccgca tacgtcaaag caaccatagt 4561 acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg 4621 ctacacttgc cagcgcctta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca 4681 cgttcgccgg ctttccccgt caagctctaa atcgggggct ccctttaggg ttccgattta 4741 gtgctttacg gcacctcgac cccaaaaaac ttgatttggg tgatggttca cgtagtgggc 4801 catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg 4861 gactcttgtt ccaaactgga acaacactca actctatctc gggctattct tttgatttat 4921 aagggatttt gccgatttcg gtctattggt taaaaaatga gctgatttaa caaaaattta 4981 acgcgaattt taacaaaata ttaacgttta caattttatg gtgcactctc agtacaatct 5041 gctctgatgc cgcatagtta agccagcccc gacacccgcc aacacccgct gacgcgccct 5101 gacgggcttg tctgctcccg gcatccgctt acagacaagc tgtgaccgtc tccgggagct 5161 gcatgtgtca gaggttttca ccgtcatcac cgaaacgcgc gagacgaaag ggcctcgtga 5221 tacgcctatt tttataggtt aatgtcatga taataatggt ttcttagacg tcaggtggca 5281 cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata 5341 tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga 5401 gtatgagtat tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc 5461 ctgtttttgc tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg 5521 cacgagtggg ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc 5581 ccgaagaacg ttttccaatg atgagcactt ttaaagttct gctatgtggc gcggtattat 5641 cccgtattga cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact 5701 tggttgagta ctcaccagtc acagaaaagc atcttacgga tggcatgaca gtaagagaat 5761 tatgcagtgc tgccataacc atgagtgata acactgcggc caacttactt ctgacaacga 5821 tcggaggacc gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc 5881 ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt gacaccacga 5941 tgcctgtagc aatggcaaca acgttgcgca aactattaac tggcgaacta cttactctag 6001 cttcccggca acaattaata gactggatgg aggcggataa agttgcagga ccacttctgc 6061 gctcggccct tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtgggt 6121 ctcgcggtat cattgcagca ctggggccag atggtaagcc ctcccgtatc gtagttatct 6181 acacgacggg gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg 6241 cctcactgat taagcattgg taactgtcag accaagttta ctcatatata ctttagattg 6301 atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca 6361 tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga 6421 tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa 6481 aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga 6541 aggtaactgg cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt 6601 taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt 6661 taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat 6721 agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct 6781 tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca 6841 cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag 6901 agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc 6961 gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga 7021 aaaacgccag caacg //