LOCUS Exported 6267 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Optogenetics. ACCESSION . VERSION . KEYWORDS pAAV-hSyn-ChRmine-mScarlet-WPRE SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6267) AUTHORS Marshel JH, Kim YS, Machado TA, Quirin S, Benson B, Kadmon J, Raja C, Chibukhchyan A, Ramakrishnan C, Inoue M, Shane JC, McKnight DJ, Yoshizawa S, Kato HE, Ganguli S, Deisseroth K TITLE Cortical layer-specific critical dynamics triggering perception. JOURNAL Science. 2019 Aug 9;365(6453). pii: science.aaw5202. doi: 10.1126/science.aaw5202. Epub 2019 Jul 18. PUBMED 31320556 REFERENCE 2 (bases 1 to 6267) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..6267 /organism="synthetic DNA construct" /mol_type="other DNA" repeat_region 6..135 /label=AAV2 ITR (alternate) /note="Functional equivalent of wild-type AAV2 ITR" promoter 180..627 /label=hSyn promoter /note="human synapsin I promoter; confers neuron-specific expression (Kugler et al., 2003)" regulatory 653..662 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 1655..2347 /codon_start=1 /product="bright monomeric red fluorescent protein evolved from a synthetic template (Bindels et al., 2016)" /label=mScarlet /translation="VSKGEAVIKEFMRFKVHMEGSMNGHEFEIEGEGEGRPYEGTQTAK LKVTKGGPLPFSWDILSPQFMYGSRAFTKHPADIPDYYKQSFPEGFKWERVMNFEDGGA VTVTQDTSLEDGTLIYKVKLRGTNFPPDGPVMQKKTMGWEASTERLYPEDGVLKGDIKM ALRLKDGGRYLADFKTTYKAKKPVQMPGAYNVDRKLDITSHNEDYTVVEQYERSEGRHS TGGMDELYK" primer_bind complement(1785..1803) /label=mCherry-R /note="mCherry, reverse primer" primer_bind 2053..2072 /label=mCherry-F /note="mCherry, forward primer" misc_feature 2399..2987 /label=WPRE /note="woodchuck hepatitis virus posttranscriptional regulatory element" primer_bind complement(2452..2472) /label=WPRE-R /note="WPRE, reverse primer" CDS complement(2870..2881) /codon_start=1 /product="Factor Xa recognition and cleavage site" /label=Factor Xa site /translation="IEGR" polyA_signal 3019..3495 /label=hGH poly(A) signal /note="human growth hormone polyadenylation signal" primer_bind complement(3218..3237) /label=hGH-PA-R /note="Human growth hormone terminator, reverse primer" repeat_region 3535..3675 /label=AAV2 ITR /note="inverted terminal repeat of adeno-associated virus serotype 2" repeat_region 3535..3664 /label=AAV2 ITR rep_origin 3750..4205 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(3837..3856) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 4047..4068 /label=F1ori-F /note="F1 origin, forward primer" primer_bind complement(4222..4241) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind 4341..4363 /label=pGEX 3' /note="pGEX vectors, reverse primer" primer_bind complement(4401..4419) /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" promoter 4487..4591 /gene="bla" /label=AmpR promoter CDS 4592..5452 /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(4810..4829) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 5623..6211 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 6112..6131 /label=pBR322ori-F /note="pBR322 origin, forward primer" ORIGIN 1 catgtcctgc aggcagctgc gcgctcgctc gctcactgag gccgcccggg cgtcgggcga 61 cctttggtcg cccggcctca gtgagcgagc gagcgcgcag agagggagtg gccaactcca 121 tcactagggg ttcctgcggc cgcacgcgtg tgtctagact gcagagggcc ctgcgtatga 181 gtgcaagtgg gttttaggac caggatgagg cggggtgggg gtgcctacct gacgaccgac 241 cccgacccac tggacaagca cccaaccccc attccccaaa ttgcgcatcc cctatcagag 301 agggggaggg gaaacaggat gcggcgaggc gcgtgcgcac tgccagcttc agcaccgcgg 361 acagtgcctt cgcccccgcc tggcggcgcg cgccaccgcc gcctcagcac tgaaggcgcg 421 ctgacgtcac tcgccggtcc cccgcaaact ccccttcccg gccaccttgg tcgcgtccgc 481 gccgccgccg gcccagccgg accgcaccac gcgaggcgcg agataggggg gcacgggcgc 541 gaccatctgc gctgcggcgc cggcgactca gcgctgcctc agtctgcggt gggcagcgga 601 ggagtcgtgt cgtgcctgag agcgcagtcg agaaggatcc ccgggtaccg gtgccaccat 661 ggcacacgca ccaggcaccg accagatgtt ctacgtgggc acaatggacg gctggtatct 721 ggataccaag ctgaactccg tggccatcgg cgcccactgg tcttgcttta tcgtgctgac 781 aatcaccaca ttctacctgg gctatgagtc ctggacatct cggggcccta gcaagagaac 841 ctccttttac gccggctatc aggaggagca gaacctggcc ctgtttgtga atttctttgc 901 catgctgagc tacttcggca agatcgtggc cgacacactg ggccacaact tcggcgatgt 961 gggccccttc atcatcggct tcggcaatta caggtatgcc gactacatgc tgacctgccc 1021 aatgctggtg tacgatctgc tgtatcagct gagggccccc tatcgcgtgt cttgtagcgc 1081 catcatcttt gccatcctga tgtctggcgt gctggccgag ttctacgcag agggcgaccc 1141 taggctgagg aatggcgcct acgcctggta tggctttggc tgtttctggt ttatcttcgc 1201 ctactctatc gtgatgagca tcgtggccaa gcagtatagc cggctggccc agctggccca 1261 ggatacaggc gccgagcaca gcctgcacgt gctgaagttc gccgtgttta ccttctccat 1321 gctgtggatt ctgtttcccc tggtgtgggc catctgccct agaggcttcg gctggatcga 1381 cgataactgg acagaggtgg cccactgcgt gtgcgacatc gtggccaagt cctgttacgg 1441 ctttgccctg gcccggttca gaaagaccta tgatgaggag ctgtttcggc tgctggagca 1501 gctgggacac gacgaggatg agttccagaa gctggagctg gatatgaggc tgagcagcaa 1561 tggcgagcgc ctgcggagac tgtctgcggc cgccaagagc aggatcacca gcgagggcga 1621 gtacatcccc ctggaccaga tcgacatcaa cgtggtgagc aagggcgagg cagtgatcaa 1681 ggagttcatg cggttcaagg tgcacatgga gggctccatg aacggccacg agttcgagat 1741 cgagggcgag ggcgagggcc gcccctacga gggcacccag accgccaagc tgaaggtgac 1801 caagggtggc cccctgccct tctcctggga catcctgtcc cctcagttca tgtacggctc 1861 cagggccttc accaagcacc ccgccgacat ccccgactac tataagcagt ccttccccga 1921 gggcttcaag tgggagcgcg tgatgaactt cgaggacggc ggcgccgtga ccgtgaccca 1981 ggacacctcc ctggaggacg gcaccctgat ctacaaggtg aagctccgcg gcaccaactt 2041 ccctcctgac ggccccgtaa tgcagaagaa gacaatgggc tgggaagcgt ccaccgagcg 2101 gttgtacccc gaggacggcg tgctgaaggg cgacattaag atggccctgc gcctgaagga 2161 cggcgggcgc tacctggcgg acttcaagac cacctacaag gccaagaagc ccgtgcagat 2221 gcccggcgcc tacaacgtgg accgcaagtt ggacatcacc tcccacaacg aggactacac 2281 cgtggtggaa cagtacgaac gctccgaggg ccgccactcc accggcggca tggacgagct 2341 gtacaagttc tgctacgaga acgaggtgta atgagaattc gatatcaagc ttatcgataa 2401 tcaacctctg gattacaaaa tttgtgaaag attgactggt attcttaact atgttgctcc 2461 ttttacgcta tgtggatacg ctgctttaat gcctttgtat catgctattg cttcccgtat 2521 ggctttcatt ttctcctcct tgtataaatc ctggttgctg tctctttatg aggagttgtg 2581 gcccgttgtc aggcaacgtg gcgtggtgtg cactgtgttt gctgacgcaa cccccactgg 2641 ttggggcatt gccaccacct gtcagctcct ttccgggact ttcgctttcc ccctccctat 2701 tgccacggcg gaactcatcg ccgcctgcct tgcccgctgc tggacagggg ctcggctgtt 2761 gggcactgac aattccgtgg tgttgtcggg gaaatcatcg tcctttcctt ggctgctcgc 2821 ctatgttgcc acctggattc tgcgcgggac gtccttctgc tacgtccctt cggccctcaa 2881 tccagcggac cttccttccc gcggcctgct gccggctctg cggcctcttc cgcgtcttcg 2941 ccttcgccct cagacgagtc ggatctccct ttgggccgcc tccccgcatc gataccgagc 3001 gctgctcgag agatctacgg gtggcatccc tgtgacccct ccccagtgcc tctcctggcc 3061 ctggaagttg ccactccagt gcccaccagc cttgtcctaa taaaattaag ttgcatcatt 3121 ttgtctgact aggtgtcctt ctataatatt atggggtgga ggggggtggt atggagcaag 3181 gggcaagttg ggaagacaac ctgtagggcc tgcggggtct attgggaacc aagctggagt 3241 gcagtggcac aatcttggct cactgcaatc tccgcctcct gggttcaagc gattctcctg 3301 cctcagcctc ccgagttgtt gggattccag gcatgcatga ccaggctcag ctaatttttg 3361 tttttttggt agagacgggg tttcaccata ttggccaggc tggtctccaa ctcctaatct 3421 caggtgatct acccaccttg gcctcccaaa ttgctgggat tacaggcgtg aaccactgct 3481 cccttccctg tccttctgat tttgtaggta accacgtgcg gaccgagcgg ccgcaggaac 3541 ccctagtgat ggagttggcc actccctctc tgcgcgctcg ctcgctcact gaggccgggc 3601 gaccaaaggt cgcccgacgc ccgggctttg cccgggcggc ctcagtgagc gagcgagcgc 3661 gcagctgcct gcaggggcgc ctgatgcggt attttctcct tacgcatctg tgcggtattt 3721 cacaccgcat acgtcaaagc aaccatagta cgcgccctgt agcggcgcat taagcgcggc 3781 gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccttag cgcccgctcc 3841 tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa 3901 tcgggggctc cctttagggt tccgatttag tgctttacgg cacctcgacc ccaaaaaact 3961 tgatttgggt gatggttcac gtagtgggcc atcgccctga tagacggttt ttcgcccttt 4021 gacgttggag tccacgttct ttaatagtgg actcttgttc caaactggaa caacactcaa 4081 ctctatctcg ggctattctt ttgatttata agggattttg ccgatttcgg tctattggtt 4141 aaaaaatgag ctgatttaac aaaaatttaa cgcgaatttt aacaaaatat taacgtttac 4201 aattttatgg tgcactctca gtacaatctg ctctgatgcc gcatagttaa gccagccccg 4261 acacccgcca acacccgctg acgcgccctg acgggcttgt ctgctcccgg catccgctta 4321 cagacaagct gtgaccgtct ccgggagctg catgtgtcag aggttttcac cgtcatcacc 4381 gaaacgcgcg agacgaaagg gcctcgtgat acgcctattt ttataggtta atgtcatgat 4441 aataatggtt tcttagacgt caggtggcac ttttcgggga aatgtgcgcg gaacccctat 4501 ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata 4561 aatgcttcaa taatattgaa aaaggaagag tatgagtatt caacatttcc gtgtcgccct 4621 tattcccttt tttgcggcat tttgccttcc tgtttttgct cacccagaaa cgctggtgaa 4681 agtaaaagat gctgaagatc agttgggtgc acgagtgggt tacatcgaac tggatctcaa 4741 cagcggtaag atccttgaga gttttcgccc cgaagaacgt tttccaatga tgagcacttt 4801 taaagttctg ctatgtggcg cggtattatc ccgtattgac gccgggcaag agcaactcgg 4861 tcgccgcata cactattctc agaatgactt ggttgagtac tcaccagtca cagaaaagca 4921 tcttacggat ggcatgacag taagagaatt atgcagtgct gccataacca tgagtgataa 4981 cactgcggcc aacttacttc tgacaacgat cggaggaccg aaggagctaa ccgctttttt 5041 gcacaacatg ggggatcatg taactcgcct tgatcgttgg gaaccggagc tgaatgaagc 5101 cataccaaac gacgagcgtg acaccacgat gcctgtagca atggcaacaa cgttgcgcaa 5161 actattaact ggcgaactac ttactctagc ttcccggcaa caattaatag actggatgga 5221 ggcggataaa gttgcaggac cacttctgcg ctcggccctt ccggctggct ggtttattgc 5281 tgataaatct ggagccggtg agcgtgggtc tcgcggtatc attgcagcac tggggccaga 5341 tggtaagccc tcccgtatcg tagttatcta cacgacgggg agtcaggcaa ctatggatga 5401 acgaaataga cagatcgctg agataggtgc ctcactgatt aagcattggt aactgtcaga 5461 ccaagtttac tcatatatac tttagattga tttaaaactt catttttaat ttaaaaggat 5521 ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt 5581 ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct 5641 gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc 5701 ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc 5761 aaatactgtt cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc 5821 gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc 5881 gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg 5941 aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata 6001 cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta 6061 tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc 6121 ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg 6181 atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt 6241 cctggccttt tgctggcctt ttgctca //