LOCUS Exported 2288 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Human expression plasmid for SaCas9 sgRNA(84nt in length) (need to clone in spacer into BsmBI sites): U6-BsmBIcassette-Sa-sgRNA(84). ACCESSION . VERSION . KEYWORDS BPK2660 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 2288) AUTHORS Kleinstiver BP, Prew MS, Tsai SQ, Nguyen NT, Topkar VV, Zheng Z, Joung JK TITLE Broadening the targeting range of Staphylococcus aureus CRISPR-Cas9 by modifying PAM recognition. JOURNAL Nat Biotechnol. 2015 Nov 2. doi: 10.1038/nbt.3404. PUBMED 26524662 REFERENCE 2 (bases 1 to 2288) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..2288 /organism="synthetic DNA construct" /mol_type="other DNA" promoter 82..322 /label=U6 promoter /note="RNA polymerase III promoter for human U6 snRNA" primer_bind 82..102 /label=hU6-F /note="Human U6 promoter, forward primer" primer_bind 253..272 /label=LKO.1 5' /note="Human U6 promoter, forward primer" rep_origin complement(539..1127) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(619..638) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(1298..2158) /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 1921..1940 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(2159..2263) /gene="bla" /label=AmpR promoter ORIGIN 1 gacgtcgcta gctgtacaaa aaagcaggct ttaaaggaac caattcagtc gactggatcc 61 ggtaccaagg tcgggcagga agagggccta tttcccatga ttccttcata tttgcatata 121 cgatacaagg ctgttagaga gataattaga attaatttga ctgtaaacac aaagatatta 181 gtacaaaata cgtgacgtag aaagtaataa tttcttgggt agtttgcagt tttaaaatta 241 tgttttaaaa tggactatca tatgcttacc gtaacttgaa agtatttcga tttcttggct 301 ttatatatct tgtggaaagg acgaaacacc cgagacgatt aatgcgtctc ggttttagta 361 ctctgtaatg aaaattacag aatctactaa aacaaggcaa aatgccgtgt ttatctcgtc 421 aacttgttgg cgagattttt ttaagcttgg gccgctcgag gtacctctct acatatgaca 481 tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt 541 tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc 601 gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct 661 ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg 721 tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca 781 agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact 841 atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta 901 acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta 961 actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct 1021 tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt 1081 tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga 1141 tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca 1201 tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat 1261 caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg 1321 cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt 1381 agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag 1441 acccacgctc accggctcca gatttatcag caataaacca gccagccgga agggccgagc 1501 gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag 1561 ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca 1621 tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa 1681 ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga 1741 tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata 1801 attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca 1861 agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg 1921 ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg 1981 ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg 2041 cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag 2101 gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac 2161 tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca 2221 tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag 2281 tgccacct //