LOCUS Exported 7421 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION AAV vector expressing AsCpf1 (RR variant). ACCESSION . VERSION . KEYWORDS hSyn1-AsCpf1(RR) SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 7421) AUTHORS Tekin, Halil, Sean Simmons, Beryl Cummings, Linyi Gao, Xian Adiconis, Cynthia C. Hession, Ayan Ghoshal, Danielle Dionne, Sourav R. Choudhury, Volkan Yesilyurt, Neville E. Sanjana, Xi Shi, Congyi TITLE Effects of 3D culturing conditions on the transcriptomic profile of stem-cell-derived neurons JOURNAL Nature Biomedical Engineering (Epub 09 April 2018) REFERENCE 2 (bases 1 to 7421) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..7421 /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 164..611 /label=hSyn promoter /note="human synapsin I promoter; confers neuron-specific expression (Kugler et al., 2003)" CDS 652..678 /codon_start=1 /product="HA (human influenza hemagglutinin) epitope tag" /label=HA /translation="YPYDVPDYA" CDS 682..702 /codon_start=1 /product="nuclear localization signal of SV40 (simian virus 40) large T antigen" /label=SV40 NLS /translation="PKKKRKV" CDS 712..4629 /codon_start=1 /product="CRISPR-associated protein from Acidaminococcus sp. BV3L6 (Zetsche et al., 2015)" /label=AsCpf1 /translation="TQFEGFTNLYQVSKTLRFELIPQGKTLKHIQEQGFIEEDKARNDH YKELKPIIDRIYKTYADQCLQLVQLDWENLSAAIDSYRKEKTEETRNALIEEQATYRNA IHDYFIGRTDNLTDAINKRHAEIYKGLFKAELFNGKVLKQLGTVTTTEHENALLRSFDK FTTYFSGFYENRKNVFSAEDISTAIPHRIVQDNFPKFKENCHIFTRLITAVPSLREHFE NVKKAIGIFVSTSIEEVFSFPFYNQLLTQTQIDLYNQLLGGISREAGTEKIKGLNEVLN LAIQKNDETAHIIASLPHRFIPLFKQILSDRNTLSFILEEFKSDEEVIQSFCKYKTLLR NENVLETAEALFNELNSIDLTHIFISHKKLETISSALCDHWDTLRNALYERRISELTGK ITKSAKEKVQRSLKHEDINLQEIISAAGKELSEAFKQKTSEILSHAHAALDQPLPTTLK KQEEKEILKSQLDSLLGLYHLLDWFAVDESNEVDPEFSARLTGIKLEMEPSLSFYNKAR NYATKKPYSVEKFKLNFQMPTLARGWDVNKEKNNGAILFVKNGLYYLGIMPKQKGRYKA LSFEPTEKTSEGFDKMYYDYFPDAAKMIPRCSTQLKAVTAHFQTHTTPILLSNNFIEPL EITKEIYDLNNPEKEPKKFQTAYAKKTGDQKGYREALCKWIDFTRDFLSKYTKTTSIDL SSLRPSSQYKDLGEYYAELNPLLYHISFQRIAEKEIMDAVETGKLYLFQIYNKDFAKGH HGKPNLHTLYWTGLFSPENLAKTSIKLNGQAELFYRPKSRMKRMAHRLGEKMLNKKLKD QKTPIPDTLYQELYDYVNHRLSHDLSDEARALLPNVITKEVSHEIIKDRRFTSDKFFFH VPITLNYQAANSPSKFNQRVNAYLKEHPETPIIGIDRGERNLIYITVIDSTGKILEQRS LNTIQQFDYQKKLDNREKERVAARQAWSVVGTIKDLKQGYLSQVIHEIVDLMIHYQAVV VLENLNFGFKSKRTGIAEKAVYQQFEKMLIDKLNCLVLKDYPAEKVGGVLNPYQLTDQF TSFAKMGTQSGFLFYVPAPYTSKIDPLTGFVDPFVWKTIKNHESRKHFLEGFDFLHYDV KTGDFILHFKMNRNLSFQRGLPGFMPAWDIVFEKNETQFDAKGTPFIAGKRIVPVIENH RFTGRYRDLYPANELIALLEEKGIVFRDGSNILPKLLENDDSHAIDTMVALIRSVLQMR NSNAATGEDYINSPVRDLNGVCFDSRFQNPEWPMDADANGAYHIALKGQLLLNHLKESK DLKLQNGISNQDWLAYIQELRN" regulatory 4349..4358 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" polyA_signal 4639..4687 /note="synthetic polyadenylation signal" repeat_region 4695..4835 /label=AAV2 ITR /note="inverted terminal repeat of adeno-associated virus serotype 2" repeat_region 4695..4824 /label=AAV2 ITR rep_origin 4910..5365 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(4997..5016) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 5207..5228 /label=F1ori-F /note="F1 origin, forward primer" primer_bind complement(5382..5401) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind 5501..5523 /label=pGEX 3' /note="pGEX vectors, reverse primer" primer_bind complement(5561..5579) /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" promoter 5647..5751 /gene="bla" /label=AmpR promoter CDS 5752..6612 /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(5970..5989) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 6783..7371 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 7272..7291 /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 gcggcctcta gactgcagag ggccctgcgt atgagtgcaa gtgggtttta 181 ggaccaggat gaggcggggt gggggtgcct acctgacgac cgaccccgac ccactggaca 241 agcacccaac ccccattccc caaattgcgc atcccctatc agagaggggg aggggaaaca 301 ggatgcggcg aggcgcgtgc gcactgccag cttcagcacc gcggacagtg ccttcgcccc 361 cgcctggcgg cgcgcgccac cgccgcctca gcactgaagg cgcgctgacg tcactcgccg 421 gtcccccgca aactcccctt cccggccacc ttggtcgcgt ccgcgccgcc gccggcccag 481 ccggaccgca ccacgcgagg cgcgagatag gggggcacgg gcgcgaccat ctgcgctgcg 541 gcgccggcga ctcagcgctg cctcagtctg cggtgggcag cggaggagtc gtgtcgtgcc 601 tgagagcgca gtcgagaagg taccggatcc ccgggtaccg gtgccaccat gtacccatac 661 gatgttccag attacgcttc gccgaagaaa aagcgcaagg tcgaagcgtc cacacagttc 721 gagggcttta ccaacctgta tcaggtgagc aagacactgc ggtttgagct gatcccacag 781 ggcaagaccc tgaagcacat ccaggagcag ggcttcatcg aggaggacaa ggcccgcaat 841 gatcactaca aggagctgaa gcccatcatc gatcggatct acaagaccta tgccgaccag 901 tgcctgcagc tggtgcagct ggattgggag aacctgagcg ccgccatcga ctcctataga 961 aaggagaaaa ccgaggagac aaggaacgcc ctgatcgagg agcaggccac atatcgcaat 1021 gccatccacg actacttcat cggccggaca gacaacctga ccgatgccat caataagaga 1081 cacgccgaga tctacaaggg cctgttcaag gccgagctgt ttaatggcaa ggtgctgaag 1141 cagctgggca ccgtgaccac aaccgagcac gagaacgccc tgctgcggag cttcgacaag 1201 tttacaacct acttctccgg cttttatgag aacaggaaga acgtgttcag cgccgaggat 1261 atcagcacag ccatcccaca ccgcatcgtg caggacaact tccccaagtt taaggagaat 1321 tgtcacatct tcacacgcct gatcaccgcc gtgcccagcc tgcgggagca ctttgagaac 1381 gtgaagaagg ccatcggcat cttcgtgagc acctccatcg aggaggtgtt ttccttccct 1441 ttttataacc agctgctgac acagacccag atcgacctgt ataaccagct gctgggagga 1501 atctctcggg aggcaggcac cgagaagatc aagggcctga acgaggtgct gaatctggcc 1561 atccagaaga atgatgagac agcccacatc atcgcctccc tgccacacag attcatcccc 1621 ctgtttaagc agatcctgtc cgataggaac accctgtctt tcatcctgga ggagtttaag 1681 agcgacgagg aagtgatcca gtccttctgc aagtacaaga cactgctgag aaacgagaac 1741 gtgctggaga cagccgaggc cctgtttaac gagctgaaca gcatcgacct gacacacatc 1801 ttcatcagcc acaagaagct ggagacaatc agcagcgccc tgtgcgacca ctgggataca 1861 ctgaggaatg ccctgtatga gcggagaatc tccgagctga caggcaagat caccaagtct 1921 gccaaggaga aggtgcagcg cagcctgaag cacgaggata tcaacctgca ggagatcatc 1981 tctgccgcag gcaaggagct gagcgaggcc ttcaagcaga aaaccagcga gatcctgtcc 2041 cacgcacacg ccgccctgga tcagccactg cctacaaccc tgaagaagca ggaggagaag 2101 gagatcctga agtctcagct ggacagcctg ctgggcctgt accacctgct ggactggttt 2161 gccgtggatg agtccaacga ggtggacccc gagttctctg cccggctgac cggcatcaag 2221 ctggagatgg agccttctct gagcttctac aacaaggcca gaaattatgc caccaagaag 2281 ccctactccg tggagaagtt caagctgaac tttcagatgc ctacactggc ccggggctgg 2341 gacgtgaata aggagaagaa caatggcgcc atcctgtttg tgaagaacgg cctgtactat 2401 ctgggcatca tgccaaagca gaagggcagg tataaggccc tcagcttcga gcccacagag 2461 aaaaccagcg agggctttga taagatgtac tatgactact ttccggatgc cgccaagatg 2521 atcccacggt gcagcaccca gctgaaggcg gtgaccgccc actttcagac ccacacaacc 2581 cccatcctgc tgtccaacaa tttcatcgag cctctggaga tcacaaagga gatctacgac 2641 ctgaacaatc ctgagaagga gccaaagaag tttcagacag cgtacgccaa gaaaaccggc 2701 gaccagaagg gctacagaga ggccctgtgc aagtggatcg acttcacaag ggattttctg 2761 tccaagtata ccaagacaac ctctatcgat ctgtctagcc tgcggccatc ctctcagtat 2821 aaggacctgg gcgagtacta tgccgagctg aatcccctgc tgtaccacat cagcttccag 2881 agaatcgccg agaaggagat catggatgcc gtggagacag gcaagctgta cctgttccag 2941 atctataaca aggactttgc caagggccac cacggcaagc ctaatctgca cacactgtat 3001 tggaccggtc tgttttctcc agagaacctg gccaagacaa gcatcaagct gaatggccag 3061 gccgagctgt tctaccgccc taagtccagg atgaagagga tggcacaccg gctgggagag 3121 aagatgctga acaagaagct gaaggatcag aaaaccccaa tccccgacac cctgtaccag 3181 gagctgtacg actatgtgaa tcacagactg tcccacgacc tgtctgatga ggccagggcc 3241 ctgctgccca acgtgatcac caaggaggtg tctcacgaga tcatcaagga taggcgcttt 3301 accagcgaca agttcttttt ccacgtgcct atcacactga actatcaggc cgccaattcc 3361 ccatctaagt tcaaccagag ggtgaatgcc tacctgaagg agcaccccga gacacctatc 3421 atcggcatcg atcggggcga gagaaacctg atctatatca cagtgatcga ctccaccggc 3481 aagatcctgg agcagcggag cctgaacacc atccagcagt ttgattacca gaagaagctg 3541 gacaacaggg agaaggagag ggtggcagca aggcaggcct ggtctgtggt gggcacaatc 3601 aaggatctga agcagggcta tctgagccag gtcatccacg agatcgtgga cctgatgatc 3661 cactaccagg ccgtggtggt gctggagaac ctgaatttcg gctttaagag caagaggacc 3721 ggcatcgccg agaaggccgt gtaccagcag ttcgagaaga tgctgatcga taagctgaat 3781 tgcctggtgc tgaaggacta tccagcagag aaagtgggag gcgtgctgaa cccataccag 3841 ctgacagacc agttcacctc ctttgccaag atgggcaccc agtctggctt cctgttttac 3901 gtgcctgccc catatacatc taagatcgat cccctgaccg gcttcgtgga ccccttcgtg 3961 tggaaaacca tcaagaatca cgagagccgc aagcacttcc tggagggctt cgactttctg 4021 cactacgacg tgaaaaccgg cgacttcatc ctgcacttta agatgaacag aaatctgtcc 4081 ttccagaggg gcctgcccgg ctttatgcct gcatgggata tcgtgttcga gaagaacgag 4141 acacagtttg acgccaaggg cacccctttc atcgccggca agagaatcgt gccagtgatc 4201 gagaatcaca gattcaccgg cagataccgg gacctgtatc ctgccaacga gctgatcgcc 4261 ctgctggagg agaagggcat cgtgttcagg gatggctcca acatcctgcc aaagctgctg 4321 gagaatgacg attctcacgc catcgacacc atggtggccc tgatccgcag cgtgctgcag 4381 atgcggaact ccaatgccgc cacaggcgag gactatatca acagccccgt gcgcgatctg 4441 aatggcgtgt gcttcgactc ccggtttcag aacccagagt ggcccatgga cgccgatgcc 4501 aatggcgcct accacatcgc cctgaagggc cagctgctgc tgaatcacct gaaggagagc 4561 aaggatctga agctgcagaa cggcatctcc aatcaggact ggctggccta catccaggag 4621 ctgcgcaact aagaattcaa taaaagatct ttattttcat tagatctgtg tgttggtttt 4681 ttgtgtgcgg ccgcaggaac ccctagtgat ggagttggcc actccctctc tgcgcgctcg 4741 ctcgctcact gaggccgggc gaccaaaggt cgcccgacgc ccgggctttg cccgggcggc 4801 ctcagtgagc gagcgagcgc gcagctgcct gcaggggcgc ctgatgcggt attttctcct 4861 tacgcatctg tgcggtattt cacaccgcat acgtcaaagc aaccatagta cgcgccctgt 4921 agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc 4981 agcgccttag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc 5041 tttccccgtc aagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg 5101 cacctcgacc ccaaaaaact tgatttgggt gatggttcac gtagtgggcc atcgccctga 5161 tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc 5221 caaactggaa caacactcaa ctctatctcg ggctattctt ttgatttata agggattttg 5281 ccgatttcgg tctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaatttt 5341 aacaaaatat taacgtttac aattttatgg tgcactctca gtacaatctg ctctgatgcc 5401 gcatagttaa gccagccccg acacccgcca acacccgctg acgcgccctg acgggcttgt 5461 ctgctcccgg catccgctta cagacaagct gtgaccgtct ccgggagctg catgtgtcag 5521 aggttttcac cgtcatcacc gaaacgcgcg agacgaaagg gcctcgtgat acgcctattt 5581 ttataggtta atgtcatgat aataatggtt tcttagacgt caggtggcac ttttcgggga 5641 aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc 5701 atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag tatgagtatt 5761 caacatttcc gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct 5821 cacccagaaa cgctggtgaa agtaaaagat gctgaagatc agttgggtgc acgagtgggt 5881 tacatcgaac tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgt 5941 tttccaatga tgagcacttt taaagttctg ctatgtggcg cggtattatc ccgtattgac 6001 gccgggcaag agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtac 6061 tcaccagtca cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct 6121 gccataacca tgagtgataa cactgcggcc aacttacttc tgacaacgat cggaggaccg 6181 aaggagctaa ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg 6241 gaaccggagc tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgtagca 6301 atggcaacaa cgttgcgcaa actattaact ggcgaactac ttactctagc ttcccggcaa 6361 caattaatag actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt 6421 ccggctggct ggtttattgc tgataaatct ggagccggtg agcgtggaag ccgcggtatc 6481 attgcagcac tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg 6541 agtcaggcaa ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt 6601 aagcattggt aactgtcaga ccaagtttac tcatatatac tttagattga tttaaaactt 6661 catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc 6721 ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct 6781 tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta 6841 ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc 6901 ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt aggccaccac 6961 ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct 7021 gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat 7081 aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg 7141 acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa 7201 gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg 7261 gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga 7321 cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc 7381 aacgcggcct ttttacggtt cctggccttt tgctcacatg t //