LOCUS Exported 5236 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Optogenetics. ACCESSION . VERSION . KEYWORDS pAAV-hSyn-mScarlet SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 5236) 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 5236) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..5236 /organism="synthetic DNA construct" /mol_type="other DNA" repeat_region 1..130 /label=AAV2 ITR (alternate) /note="Functional equivalent of wild-type AAV2 ITR" promoter 175..622 /label=hSyn promoter /note="human synapsin I promoter; confers neuron-specific expression (Kugler et al., 2003)" regulatory 636..645 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 642..1337 /codon_start=1 /product="bright monomeric red fluorescent protein evolved from a synthetic template (Bindels et al., 2016)" /label=mScarlet /translation="MVSKGEAVIKEFMRFKVHMEGSMNGHEFEIEGEGEGRPYEGTQTA KLKVTKGGPLPFSWDILSPQFMYGSRAFTKHPADIPDYYKQSFPEGFKWERVMNFEDGG AVTVTQDTSLEDGTLIYKVKLRGTNFPPDGPVMQKKTMGWEASTERLYPEDGVLKGDIK MALRLKDGGRYLADFKTTYKAKKPVQMPGAYNVDRKLDITSHNEDYTVVEQYERSEGRH STGGMDELYK" primer_bind complement(775..793) /label=mCherry-R /note="mCherry, reverse primer" primer_bind 1043..1062 /label=mCherry-F /note="mCherry, forward primer" misc_feature 1363..1951 /label=WPRE /note="woodchuck hepatitis virus posttranscriptional regulatory element" primer_bind complement(1416..1436) /label=WPRE-R /note="WPRE, reverse primer" CDS complement(1834..1845) /codon_start=1 /product="Factor Xa recognition and cleavage site" /label=Factor Xa site /translation="IEGR" polyA_signal 1983..2459 /label=hGH poly(A) signal /note="human growth hormone polyadenylation signal" primer_bind complement(2182..2201) /label=hGH-PA-R /note="Human growth hormone terminator, reverse primer" repeat_region 2499..2639 /label=AAV2 ITR /note="inverted terminal repeat of adeno-associated virus serotype 2" repeat_region 2499..2628 /label=AAV2 ITR rep_origin 2714..3169 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(2801..2820) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 3011..3032 /label=F1ori-F /note="F1 origin, forward primer" primer_bind complement(3186..3205) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind 3305..3327 /label=pGEX 3' /note="pGEX vectors, reverse primer" primer_bind complement(3365..3383) /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" promoter 3451..3555 /gene="bla" /label=AmpR promoter CDS 3556..4416 /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(3774..3793) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 4587..5175 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 5076..5095 /label=pBR322ori-F /note="pBR322 origin, forward primer" ORIGIN 1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcgtcg ggcgaccttt 61 ggtcgcccgg cctcagtgag cgagcgagcg cgcagagagg gagtggccaa ctccatcact 121 aggggttcct gcggccgcac gcgtgtgtct agactgcaga gggccctgcg tatgagtgca 181 agtgggtttt aggaccagga tgaggcgggg tgggggtgcc tacctgacga ccgaccccga 241 cccactggac aagcacccaa cccccattcc ccaaattgcg catcccctat cagagagggg 301 gaggggaaac aggatgcggc gaggcgcgtg cgcactgcca gcttcagcac cgcggacagt 361 gccttcgccc ccgcctggcg gcgcgcgcca ccgccgcctc agcactgaag gcgcgctgac 421 gtcactcgcc ggtcccccgc aaactcccct tcccggccac cttggtcgcg tccgcgccgc 481 cgccggccca gccggaccgc accacgcgag gcgcgagata ggggggcacg ggcgcgacca 541 tctgcgctgc ggcgccggcg actcagcgct gcctcagtct gcggtgggca gcggaggagt 601 cgtgtcgtgc ctgagagcgc agtcgagaag gatccgccac catggtgagc aagggcgagg 661 cagtgatcaa ggagttcatg cggttcaagg tgcacatgga gggctccatg aacggccacg 721 agttcgagat cgagggcgag ggcgagggcc gcccctacga gggcacccag accgccaagc 781 tgaaggtgac caagggtggc cccctgccct tctcctggga catcctgtcc cctcagttca 841 tgtacggctc cagggccttc accaagcacc ccgccgacat ccccgactac tataagcagt 901 ccttccccga gggcttcaag tgggagcgcg tgatgaactt cgaggacggc ggcgccgtga 961 ccgtgaccca ggacacctcc ctggaggacg gcaccctgat ctacaaggtg aagctccgcg 1021 gcaccaactt ccctcctgac ggccccgtaa tgcagaagaa gacaatgggc tgggaagcgt 1081 ccaccgagcg gttgtacccc gaggacggcg tgctgaaggg cgacattaag atggccctgc 1141 gcctgaagga cggcgggcgc tacctggcgg acttcaagac cacctacaag gccaagaagc 1201 ccgtgcagat gcccggcgcc tacaacgtgg accgcaagtt ggacatcacc tcccacaacg 1261 aggactacac cgtggtggaa cagtacgaac gctccgaggg ccgccactcc accggcggca 1321 tggacgagct gtacaagtga attcgatatc aagcttatcg ataatcaacc tctggattac 1381 aaaatttgtg aaagattgac tggtattctt aactatgttg ctccttttac gctatgtgga 1441 tacgctgctt taatgccttt gtatcatgct attgcttccc gtatggcttt cattttctcc 1501 tccttgtata aatcctggtt gctgtctctt tatgaggagt tgtggcccgt tgtcaggcaa 1561 cgtggcgtgg tgtgcactgt gtttgctgac gcaaccccca ctggttgggg cattgccacc 1621 acctgtcagc tcctttccgg gactttcgct ttccccctcc ctattgccac ggcggaactc 1681 atcgccgcct gccttgcccg ctgctggaca ggggctcggc tgttgggcac tgacaattcc 1741 gtggtgttgt cggggaaatc atcgtccttt ccttggctgc tcgcctatgt tgccacctgg 1801 attctgcgcg ggacgtcctt ctgctacgtc ccttcggccc tcaatccagc ggaccttcct 1861 tcccgcggcc tgctgccggc tctgcggcct cttccgcgtc ttcgccttcg ccctcagacg 1921 agtcggatct ccctttgggc cgcctccccg catcgatacc gagcgctgct cgagagatct 1981 acgggtggca tccctgtgac ccctccccag tgcctctcct ggccctggaa gttgccactc 2041 cagtgcccac cagccttgtc ctaataaaat taagttgcat cattttgtct gactaggtgt 2101 ccttctataa tattatgggg tggagggggg tggtatggag caaggggcaa gttgggaaga 2161 caacctgtag ggcctgcggg gtctattggg aaccaagctg gagtgcagtg gcacaatctt 2221 ggctcactgc aatctccgcc tcctgggttc aagcgattct cctgcctcag cctcccgagt 2281 tgttgggatt ccaggcatgc atgaccaggc tcagctaatt tttgtttttt tggtagagac 2341 ggggtttcac catattggcc aggctggtct ccaactccta atctcaggtg atctacccac 2401 cttggcctcc caaattgctg ggattacagg cgtgaaccac tgctcccttc cctgtccttc 2461 tgattttgta ggtaaccacg tgcggaccga gcggccgcag gaacccctag tgatggagtt 2521 ggccactccc tctctgcgcg ctcgctcgct cactgaggcc gggcgaccaa aggtcgcccg 2581 acgcccgggc tttgcccggg cggcctcagt gagcgagcga gcgcgcagct gcctgcaggg 2641 gcgcctgatg cggtattttc tccttacgca tctgtgcggt atttcacacc gcatacgtca 2701 aagcaaccat agtacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg 2761 cgcagcgtga ccgctacact tgccagcgcc ttagcgcccg ctcctttcgc tttcttccct 2821 tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta 2881 gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgattt gggtgatggt 2941 tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg 3001 ttctttaata gtggactctt gttccaaact ggaacaacac tcaactctat ctcgggctat 3061 tcttttgatt tataagggat tttgccgatt tcggtctatt ggttaaaaaa tgagctgatt 3121 taacaaaaat ttaacgcgaa ttttaacaaa atattaacgt ttacaatttt atggtgcact 3181 ctcagtacaa tctgctctga tgccgcatag ttaagccagc cccgacaccc gccaacaccc 3241 gctgacgcgc cctgacgggc ttgtctgctc ccggcatccg cttacagaca agctgtgacc 3301 gtctccggga gctgcatgtg tcagaggttt tcaccgtcat caccgaaacg cgcgagacga 3361 aagggcctcg tgatacgcct atttttatag gttaatgtca tgataataat ggtttcttag 3421 acgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa 3481 atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat 3541 tgaaaaagga agagtatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg 3601 gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa 3661 gatcagttgg gtgcacgagt gggttacatc gaactggatc tcaacagcgg taagatcctt 3721 gagagttttc gccccgaaga acgttttcca atgatgagca cttttaaagt tctgctatgt 3781 ggcgcggtat tatcccgtat tgacgccggg caagagcaac tcggtcgccg catacactat 3841 tctcagaatg acttggttga gtactcacca gtcacagaaa agcatcttac ggatggcatg 3901 acagtaagag aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta 3961 cttctgacaa cgatcggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat 4021 catgtaactc gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag 4081 cgtgacacca cgatgcctgt agcaatggca acaacgttgc gcaaactatt aactggcgaa 4141 ctacttactc tagcttcccg gcaacaatta atagactgga tggaggcgga taaagttgca 4201 ggaccacttc tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc 4261 ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt 4321 atcgtagtta tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc 4381 gctgagatag gtgcctcact gattaagcat tggtaactgt cagaccaagt ttactcatat 4441 atactttaga ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt 4501 tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac 4561 cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc 4621 ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca 4681 actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta 4741 gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct 4801 ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg 4861 gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc 4921 acacagccca gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta 4981 tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg 5041 gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt 5101 cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg 5161 cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg 5221 ccttttgctc acatgt //