LOCUS Exported 6980 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION AAV expression of CaMKIIa-driven Arch 3.0 for optical inhibition.. ACCESSION . VERSION . KEYWORDS pAAV-CaMKIIa-eArch 3.0-EYFP SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6980) AUTHORS Mattis J, Tye KM, Ferenczi EA, Ramakrishnan C, O'Shea DJ, Prakash R, Gunaydin LA, Hyun M, Fenno LE, Gradinaru V, Yizhar O, Deisseroth K TITLE Principles for applying optogenetic tools derived from direct comparative analysis of microbial opsins. JOURNAL Nat Methods. 2011 Dec 18;9(2):159-72. doi: 10.1038/nmeth.1808. PUBMED 22179551 REFERENCE 2 (bases 1 to 6980) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..6980 /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 160..1448 /label=CaMKII promoter /note="promoter for the mouse calcium/calmodulin-dependent kinase II alpha subunit" regulatory 1469..1478 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 1475..2248 /codon_start=1 /product="light-driven proton pump archaerhodopsin-3 from Halorubrum sodomense (Chow et al., 2010)" /label=Arch /translation="MDPIALQAGYDLLGDGRPETLWLGIGTLLMLIGTFYFLVRGWGVT DKDAREYYAVTILVPGIASAAYLSMFFGIGLTEVTVGGEMLDIYYARYADWLFTTPLLL LDLALLAKVDRVTIGTLVGVDALMIVTGLIGALSHTAIARYSWWLFSTICMIVVLYFLA TSLRSAAKERGPEVASTFNTLTALVLVLWTAYPILWIIGTEGAGVVGLGIETLLFMVLD VTAKVGFGFILLRSRAILGDTEAPEPSAGADVSAAD" CDS 2342..3055 /codon_start=1 /product="enhanced YFP" /label=EYFP /note="mammalian codon-optimized" /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK FICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLLE FVTAAGITLGMDELYK" primer_bind complement(2384..2405) /label=EGFP-N /note="EGFP, reverse primer" primer_bind complement(2645..2664) /label=EXFP-R /note="For distinguishing EGFP variants, reverse primer" primer_bind 2992..3013 /label=EGFP-C /note="EGFP, forward primer" misc_feature 3107..3695 /label=WPRE /note="woodchuck hepatitis virus posttranscriptional regulatory element" primer_bind complement(3160..3180) /label=WPRE-R /note="WPRE, reverse primer" CDS complement(3578..3589) /codon_start=1 /product="Factor Xa recognition and cleavage site" /label=Factor Xa site /translation="IEGR" polyA_signal 3727..4203 /label=hGH poly(A) signal /note="human growth hormone polyadenylation signal" primer_bind complement(3926..3945) /label=hGH-PA-R /note="Human growth hormone terminator, reverse primer" repeat_region 4243..4383 /label=AAV2 ITR /note="inverted terminal repeat of adeno-associated virus serotype 2" repeat_region 4243..4372 /label=AAV2 ITR rep_origin 4458..4913 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(4545..4564) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 4755..4776 /label=F1ori-F /note="F1 origin, forward primer" primer_bind complement(4930..4949) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind 5049..5071 /label=pGEX 3' /note="pGEX vectors, reverse primer" primer_bind complement(5109..5127) /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" promoter 5195..5299 /gene="bla" /label=AmpR promoter CDS 5300..6160 /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(5518..5537) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 6331..6919 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 6820..6839 /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 cgcgtttaac attatggcct taggtcactt 181 catctccatg gggttcttct tctgattttc tagaaaatga gatgggggtg cagagagctt 241 cctcagtgac ctgcccaggg tcacatcaga aatgtcagag ctagaacttg aactcagatt 301 actaatctta aattccatgc cttgggggca tgcaagtacg atatacagaa ggagtgaact 361 cattagggca gatgaccaat gagtttagga aagaagagtc cagggcaggg tacatctaca 421 ccacccgccc agccctgggt gagtccagcc acgttcacct cattatagtt gcctctctcc 481 agtcctacct tgacgggaag cacaagcaga aactgggaca ggagccccag gagaccaaat 541 cttcatggtc cctctgggag gatgggtggg gagagctgtg gcagaggcct caggaggggc 601 cctgctgctc agtggtgaca gataggggtg agaaagcaga cagagtcatt ccgtcagcat 661 tctgggtctg tttggtactt cttctcacgc taaggtggcg gtgtgatatg cacaatggct 721 aaaaagcagg gagagctgga aagaaacaag gacagagaca gaggccaagt caaccagacc 781 aattcccaga ggaagcaaag aaaccattac agagactaca agggggaagg gaaggagaga 841 tgaattagct tcccctgtaa accttagaac ccagctgttg ccagggcaac ggggcaatac 901 ctgtctcttc agaggagatg aagttgccag ggtaactaca tcctgtcttt ctcaaggacc 961 atcccagaat gtggcaccca ctagccgtta ccatagcaac tgcctctttg ccccacttaa 1021 tcccatcccg tctgttaaaa gggccctata gttggaggtg ggggaggtag gaagagcgat 1081 gatcacttgt ggactaagtt tgttcgcatc cccttctcca accccctcag tacatcaccc 1141 tgggggaaca gggtccactt gctcctgggc ccacacagtc ctgcagtatt gtgtatataa 1201 ggccagggca aagaggagca ggttttaaag tgaaaggcag gcaggtgttg gggaggcagt 1261 taccggggca acgggaacag ggcgtttcgg aggtggttgc catggggacc tggatgctga 1321 cgaaggctcg cgaggctgtg agcagccaca gtgccctgct cagaagcccc aagctcgtca 1381 gtcaagccgg ttctccgttt gcactcagga gcacgggcag gcgagtggcc cctagttctg 1441 ggggcagctc tagagcggta ccggatccgc caccatggac cccatcgctc tgcaggctgg 1501 ttacgacctg ctgggtgacg gcagacctga aactctgtgg ctgggcatcg gcactctgct 1561 gatgctgatt ggaaccttct actttctggt ccgcggatgg ggagtcaccg ataaggatgc 1621 ccgggaatat tacgctgtga ctatcctggt gcccggaatc gcatccgccg catatctgtc 1681 tatgttcttt ggtatcgggc ttactgaggt gaccgtcggg ggcgaaatgt tggatatcta 1741 ttatgccagg tacgccgact ggctgtttac caccccactt ctgctgctgg atctggccct 1801 tctcgctaag gtggatcggg tgaccatcgg caccctggtg ggtgtggacg ccctgatgat 1861 cgtcactggc ctcatcggag ccttgagcca cacggccata gccagataca gttggtggtt 1921 gttctctaca atttgcatga tagtggtgct ctattttctg gctacatccc tgcgatctgc 1981 tgcaaaggag cggggccccg aggtggcatc tacctttaac accctgacag ctctggtctt 2041 ggtgctgtgg accgcttacc ctatcctgtg gatcataggc actgagggcg ctggcgtggt 2101 gggcctgggc atcgaaactc tgctgtttat ggtgttggac gtgactgcca aggtcggctt 2161 tggctttatc ctgttgagat cccgggctat tctgggcgac accgaggcac cagaacccag 2221 tgccggtgcc gatgtcagtg ccgccgaccg accggtagta gcagtgagca aggcggccgc 2281 caagagcagg atcaccagcg agggcgagta catccccctg gaccagatcg acatcaacgt 2341 ggtgagcaag ggcgaggagc tgttcaccgg ggtggtgccc atcctggtcg agctggacgg 2401 cgacgtaaac ggccacaagt tcagcgtgtc cggcgagggc gagggcgatg ccacctacgg 2461 caagctgacc ctgaagttca tctgcaccac cggcaagctg cccgtgccct ggcccaccct 2521 cgtgaccacc ttcggctacg gcctgcagtg cttcgcccgc taccccgacc acatgaagca 2581 gcacgacttc ttcaagtccg ccatgcccga aggctacgtc caggagcgca ccatcttctt 2641 caaggacgac ggcaactaca agacccgcgc cgaggtgaag ttcgagggcg acaccctggt 2701 gaaccgcatc gagctgaagg gcatcgactt caaggaggac ggcaacatcc tggggcacaa 2761 gctggagtac aactacaaca gccacaacgt ctatatcatg gccgacaagc agaagaacgg 2821 catcaaggtg aacttcaaga tccgccacaa catcgaggac ggcagcgtgc agctcgccga 2881 ccactaccag cagaacaccc ccatcggcga cggccccgtg ctgctgcccg acaaccacta 2941 cctgagctac cagtccgccc tgagcaaaga ccccaacgag aagcgcgatc acatggtcct 3001 gctggagttc gtgaccgccg ccgggatcac tctcggcatg gacgagctgt acaagttctg 3061 ctacgagaac gaggtgtaat gagaattcga tatcaagctt atcgataatc aacctctgga 3121 ttacaaaatt tgtgaaagat tgactggtat tcttaactat gttgctcctt ttacgctatg 3181 tggatacgct gctttaatgc ctttgtatca tgctattgct tcccgtatgg ctttcatttt 3241 ctcctccttg tataaatcct ggttgctgtc tctttatgag gagttgtggc ccgttgtcag 3301 gcaacgtggc gtggtgtgca ctgtgtttgc tgacgcaacc cccactggtt ggggcattgc 3361 caccacctgt cagctccttt ccgggacttt cgctttcccc ctccctattg ccacggcgga 3421 actcatcgcc gcctgccttg cccgctgctg gacaggggct cggctgttgg gcactgacaa 3481 ttccgtggtg ttgtcgggga aatcatcgtc ctttccttgg ctgctcgcct gtgttgccac 3541 ctggattctg cgcgggacgt ccttctgcta cgtcccttcg gccctcaatc cagcggacct 3601 tccttcccgc ggcctgctgc cggctctgcg gcctcttccg cgtcttcgcc ttcgccctca 3661 gacgagtcgg atctcccttt gggccgcctc cccgcatcga taccgagcgc tgctcgagag 3721 atctacgggt ggcatccctg tgacccctcc ccagtgcctc tcctggccct ggaagttgcc 3781 actccagtgc ccaccagcct tgtcctaata aaattaagtt gcatcatttt gtctgactag 3841 gtgtccttct ataatattat ggggtggagg ggggtggtat ggagcaaggg gcaagttggg 3901 aagacaacct gtagggcctg cggggtctat tgggaaccaa gctggagtgc agtggcacaa 3961 tcttggctca ctgcaatctc cgcctcctgg gttcaagcga ttctcctgcc tcagcctccc 4021 gagttgttgg gattccaggc atgcatgacc aggctcagct aatttttgtt tttttggtag 4081 agacggggtt tcaccatatt ggccaggctg gtctccaact cctaatctca ggtgatctac 4141 ccaccttggc ctcccaaatt gctgggatta caggcgtgaa ccactgctcc cttccctgtc 4201 cttctgattt tgtaggtaac cacgtgcgga ccgagcggcc gcaggaaccc ctagtgatgg 4261 agttggccac tccctctctg cgcgctcgct cgctcactga ggccgggcga ccaaaggtcg 4321 cccgacgccc gggctttgcc cgggcggcct cagtgagcga gcgagcgcgc agctgcctgc 4381 aggggcgcct gatgcggtat tttctcctta cgcatctgtg cggtatttca caccgcatac 4441 gtcaaagcaa ccatagtacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt 4501 tacgcgcagc gtgaccgcta cacttgccag cgccctagcg cccgctcctt tcgctttctt 4561 cccttccttt ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc gggggctccc 4621 tttagggttc cgatttagtg ctttacggca cctcgacccc aaaaaacttg atttgggtga 4681 tggttcacgt agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc 4741 cacgttcttt aatagtggac tcttgttcca aactggaaca acactcaacc ctatctcggg 4801 ctattctttt gatttataag ggattttgcc gatttcggcc tattggttaa aaaatgagct 4861 gatttaacaa aaatttaacg cgaattttaa caaaatatta acgtttacaa ttttatggtg 4921 cactctcagt acaatctgct ctgatgccgc atagttaagc cagccccgac acccgccaac 4981 acccgctgac gcgccctgac gggcttgtct gctcccggca tccgcttaca gacaagctgt 5041 gaccgtctcc gggagctgca tgtgtcagag gttttcaccg tcatcaccga aacgcgcgag 5101 acgaaagggc ctcgtgatac gcctattttt ataggttaat gtcatgataa taatggtttc 5161 ttagacgtca ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt 5221 ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata 5281 atattgaaaa aggaagagta tgagtattca acatttccgt gtcgccctta ttcccttttt 5341 tgcggcattt tgccttcctg tttttgctca cccagaaacg ctggtgaaag taaaagatgc 5401 tgaagatcag ttgggtgcac gagtgggtta catcgaactg gatctcaaca gcggtaagat 5461 ccttgagagt tttcgccccg aagaacgttt tccaatgatg agcactttta aagttctgct 5521 atgtggcgcg gtattatccc gtattgacgc cgggcaagag caactcggtc gccgcataca 5581 ctattctcag aatgacttgg ttgagtactc accagtcaca gaaaagcatc ttacggatgg 5641 catgacagta agagaattat gcagtgctgc cataaccatg agtgataaca ctgcggccaa 5701 cttacttctg acaacgatcg gaggaccgaa ggagctaacc gcttttttgc acaacatggg 5761 ggatcatgta actcgccttg atcgttggga accggagctg aatgaagcca taccaaacga 5821 cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg 5881 cgaactactt actctagctt cccggcaaca attaatagac tggatggagg cggataaagt 5941 tgcaggacca cttctgcgct cggcccttcc ggctggctgg tttattgctg ataaatctgg 6001 agccggtgag cgtgggtctc gcggtatcat tgcagcactg gggccagatg gtaagccctc 6061 ccgtatcgta gttatctaca cgacggggag tcaggcaact atggatgaac gaaatagaca 6121 gatcgctgag ataggtgcct cactgattaa gcattggtaa ctgtcagacc aagtttactc 6181 atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat 6241 cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc 6301 agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg 6361 ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct 6421 accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa atactgtcct 6481 tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct 6541 cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg 6601 gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc 6661 gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga 6721 gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg 6781 cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta 6841 tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg 6901 ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg 6961 ctggcctttt gctcacatgt //