LOCUS Exported 992 bp ds-DNA linear SYN 17-MAY-2021 DEFINITION An empty gRNA expression vector, used to create a gRNA to a specific sequence.. ACCESSION . VERSION . KEYWORDS gRNA_Cloning Vector SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 992) AUTHORS Mali P, Yang L, Esvelt KM, Aach J, Guell M, Dicarlo JE, Norville JE, Church GM TITLE RNA-Guided Human Genome Engineering via Cas9. JOURNAL Science. 2013 Jan 3. PUBMED 23287722 REFERENCE 2 (bases 1 to 992) AUTHORS . TITLE Direct Submission JOURNAL Exported May 17, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..992 /organism="synthetic DNA construct" /mol_type="other DNA" primer_bind 115..135 /label=hU6-F /note="Human U6 promoter, forward primer" primer_bind 286..305 /label=LKO.1 5' /note="Human U6 promoter, forward primer" protein_bind complement(412..436) /gene="mutant version of attB" /label=attB2 /bound_moiety="BP Clonase(TM)" /note="recombination site for the Gateway(R) BP reaction" promoter complement(521..539) /label=SP6 promoter /note="promoter for bacteriophage SP6 RNA polymerase" primer_bind complement(522..539) /label=SP6 /note="SP6 promoter, forward primer" primer_bind complement(557..573) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(557..573) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(570..592) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 581..597 /label=lac operator /bound_moiety="lac repressor encoded by lacI" /note="The lac repressor binds to the lac operator to inhibit transcription in E. coli. This inhibition can be relieved by adding lactose or isopropyl-beta-D-thiogalactopyranoside (IPTG)." promoter complement(605..635) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 650..671 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(788..805) /label=L4440 /note="L4440 vector, forward primer" ORIGIN 1 ctcgagcggc cgccagtgtg atggatatct gcagaattcg ccctttgtac aaaaaagcag 61 gctttaaagg aaccaattca gtcgactgga tccggtacca aggtcgggca ggaagagggc 121 ctatttccca tgattccttc atatttgcat atacgataca aggctgttag agagataatt 181 agaattaatt tgactgtaaa cacaaagata ttagtacaaa atacgtgacg tagaaagtaa 241 taatttcttg ggtagtttgc agttttaaaa ttatgtttta aaatggacta tcatatgctt 301 accgtaactt gaaagtattt cgatttcttg gctttatata tcttaagtta aaataaggct 361 agtccgttat caacttgaaa aagtggcacc gagtcggtgc tttttttcta gacccagctt 421 tcttgtacaa agttggcatt aaagggcgaa ttccagcaca ctggcggccg ttactagtgg 481 atccgagctc ggtaccaagc ttgatgcata gcttgagtat tctatagtgt cacctaaata 541 gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc 601 cacacaacat acgagccgga agcataaagt gtaaagcctg gggtgcctaa tgagtgagct 661 aactcacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc 721 agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt 781 ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag 841 ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca 901 tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt 961 ccataggctc cgcccccctg acgagcatca ca //