LOCUS Exported 2278 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION guide RNA (gRNA) expression vector used to create a gRNA to a specific sequence, uses U6 promoter. ACCESSION . VERSION . KEYWORDS MLM3636 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 2278) AUTHORS Joung JK TITLE Joung lab unpublished CRISPR plasmids JOURNAL Unpublished REFERENCE 2 (bases 1 to 2278) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..2278 /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" misc_RNA 350..425 /label=gRNA scaffold /note="guide RNA scaffold for the Streptococcus pyogenes CRISPR/Cas9 system" rep_origin complement(529..1117) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(609..628) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(1288..2148) /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 1911..1930 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(2149..2253) /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 gagacgatta atgcgtctcg ttttagagct 361 agaaatagca agttaaaata aggctagtcc gttatcaact tgaaaaagtg gcaccgagtc 421 ggtgcttttt ttaagcttgg gccgctcgag gtacctctct acatatgaca tgtgagcaaa 481 aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct 541 ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac 601 aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc 661 gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc 721 tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg 781 tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga 841 gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta acaggattag 901 cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta 961 cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag 1021 agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg 1081 caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac 1141 ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc 1201 aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag 1261 tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc 1321 agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac 1381 gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc 1441 accggctcca gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg 1501 tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag 1561 tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc 1621 acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac 1681 atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag 1741 aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac 1801 tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg 1861 agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc 1921 gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact 1981 ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg 2041 atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa 2101 tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt 2161 tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg 2221 tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacct //