LOCUS Exported 329 bp ds-DNA linear SYN 17-MAY-2021 DEFINITION Expresses Cas9 nuclease and gRNA. ACCESSION . VERSION . KEYWORDS pX330A-1x7 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 329) AUTHORS Sakuma T, Nishikawa A, Kume S, Chayama K, Yamamoto T TITLE Multiplex genome engineering in human cells using all-in-one CRISPR/Cas9 vector system. JOURNAL Sci Rep. 2014 Jun 23;4:5400. doi: 10.1038/srep05400. PUBMED 24954249 REFERENCE 2 (bases 1 to 329) AUTHORS . TITLE Direct Submission JOURNAL Exported May 17, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..329 /organism="synthetic DNA construct" /mol_type="other DNA" primer_bind 2..19 /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind 3..19 /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" primer_bind 29..48 /label=T7 /note="T7 promoter, forward primer" promoter 29..47 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" primer_bind complement(95..115) /label=T3 /note="T3 promoter, forward primer" promoter complement(95..113) /label=T3 promoter /note="promoter for bacteriophage T3 RNA polymerase" primer_bind complement(134..150) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(134..150) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(147..169) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 158..174 /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(182..212) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 227..248 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." ORIGIN 1 ttgtaaaacg acggccagtg agcgcgcgta atacgactca ctatagggcg aattgggtac 61 cgggcccccc ctcgaggtcc tccagctttt gttcccttta gtgagggtta attgcgcgct 121 tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac 181 acaacatacg agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac 241 tcacattaat tgcgttgcgc tcactgcccg ctttccaccg gtggtctcta ggtacccgtt 301 acataactta cggtaaatgg cccgcctgg //