LOCUS Exported 2766 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Backbone for cloning LwCas13a guides under tRNAval promoter. ACCESSION . VERSION . KEYWORDS pC017 - LwCas13a guide expression backbone with tRNAval promoter SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 2766) AUTHORS Abudayyeh OO, Gootenberg JS, Essletzbichler P, Han S, Joung J, Belanto JJ, Verdine V, Cox DBT, Kellner MJ, Regev A, Lander ES, Voytas DF, Ting AY, Zhang F TITLE RNA targeting with CRISPR-Cas13. JOURNAL Nature. 2017 Oct 4. doi: 10.1038/nature24049. PUBMED 28976959 REFERENCE 2 (bases 1 to 2766) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..2766 /organism="synthetic DNA construct" /mol_type="other DNA" rep_origin 159..747 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 648..667 /label=pBR322ori-F /note="pBR322 origin, forward primer" rep_origin 1052..1507 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(1139..1158) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 1349..1370 /label=F1ori-F /note="F1 origin, forward primer" primer_bind complement(1524..1543) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind 1643..1665 /label=pGEX 3' /note="pGEX vectors, reverse primer" primer_bind complement(1703..1721) /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" promoter 1789..1893 /gene="bla" /label=AmpR promoter CDS 1894..2754 /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(2112..2131) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" ORIGIN 1 gtttactcat atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag 61 gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac 121 tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc 181 gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat 241 caagagctac caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat 301 actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct 361 acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt 421 cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg 481 gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta 541 cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg 601 gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg 661 tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc 721 tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg 781 gccttttgct ggccttttgc tcagctagcc aggactagtc ttttaggtca aaaagaagaa 841 gctttgtaac cgttggtttc cgtagtgtag tggttatcac gttcgcctaa cacgcgaaag 901 gtccccggtt cgaaggattt agactacccc aaaaacgaag gggactaaaa cgggtcttcg 961 agaagacctt tttttttgaa ttctgatgcg gtattttctc cttacgcatc tgtgcggtat 1021 ttcacaccgc atacgtcaaa gcaaccatag tacgcgccct gtagcggcgc attaagcgcg 1081 gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgcctt agcgcccgct 1141 cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta 1201 aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa 1261 cttgatttgg gtgatggttc acgtagtggg ccatcgccct gatagacggt ttttcgccct 1321 ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg aacaacactc 1381 aactctatct cgggctattc ttttgattta taagggattt tgccgatttc ggtctattgg 1441 ttaaaaaatg agctgattta acaaaaattt aacgcgaatt ttaacaaaat attaacgttt 1501 acaattttat ggtgcactct cagtacaatc tgctctgatg ccgcatagtt aagccagccc 1561 cgacacccgc caacacccgc tgacgcgccc tgacgggctt gtctgctccc ggcatccgct 1621 tacagacaag ctgtgaccgt ctccgggagc tgcatgtgtc agaggttttc accgtcatca 1681 ccgaaacgcg cgagacgaaa gggcctcgtg atacgcctat ttttataggt taatgtcatg 1741 ataataatgg tttcttagac gtcaggtggc acttttcggg gaaatgtgcg cggaacccct 1801 atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga 1861 taaatgcttc aataatattg aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc 1921 cttattccct tttttgcggc attttgcctt cctgtttttg ctcacccaga aacgctggtg 1981 aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg gttacatcga actggatctc 2041 aacagcggta agatccttga gagttttcgc cccgaagaac gttttccaat gatgagcact 2101 tttaaagttc tgctatgtgg cgcggtatta tcccgtattg acgccgggca agagcaactc 2161 ggtcgccgca tacactattc tcagaatgac ttggttgagt actcaccagt cacagaaaag 2221 catcttacgg atggcatgac agtaagagaa ttatgcagtg ctgccataac catgagtgat 2281 aacactgcgg ccaacttact tctgacaacg atcggaggac cgaaggagct aaccgctttt 2341 ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa 2401 gccataccaa acgacgagcg tgacaccacg atgcctgtag caatggcaac aacgttgcgc 2461 aaactattaa ctggcgaact acttactcta gcttcccggc aacaattaat agactggatg 2521 gaggcggata aagttgcagg accacttctg cgctcggccc ttccggctgg ctggtttatt 2581 gctgataaat ctggagccgg tgagcgtgga agccgcggta tcattgcagc actggggcca 2641 gatggtaagc cctcccgtat cgtagttatc tacacgacgg ggagtcaggc aactatggat 2701 gaacgaaata gacagatcgc tgagataggt gcctcactga ttaagcattg gtaactgtca 2761 gaccaa //