LOCUS Exported 868 bp ds-DNA linear SYN 18-MAY-2021 DEFINITION Expresses the S. pyogenes sgRNA from the 7SK promoter. ACCESSION . VERSION . KEYWORDS ph7SK-gRNA SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 868) AUTHORS Kabadi AM, Ousterout DG, Hilton IB, Gersbach CA TITLE Multiplex CRISPR/Cas9-based genome engineering from a single lentiviral vector. JOURNAL Nucleic Acids Res. 2014 Aug 13. pii: gku749. PUBMED 25122746 REFERENCE 2 (bases 1 to 868) AUTHORS . TITLE Direct Submission JOURNAL Exported May 18, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..868 /organism="synthetic DNA construct" /mol_type="other DNA" misc_RNA 321..396 /label=gRNA scaffold /note="guide RNA scaffold for the Streptococcus pyogenes CRISPR/Cas9 system" primer_bind complement(452..472) /label=T3 /note="T3 promoter, forward primer" promoter complement(452..470) /label=T3 promoter /note="promoter for bacteriophage T3 RNA polymerase" primer_bind complement(491..507) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(491..507) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(504..526) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 515..531 /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(539..569) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 584..605 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(722..739) /label=L4440 /note="L4440 vector, forward primer" ORIGIN 1 gactcactat agggcgaatt gggtaccttt aattctagta ctatgcatcg tctcattgtc 61 tgcagtattt agcatgcccc acccatctgc aaggcattct ggatagtgtc aaaacagccg 121 gaaatcaagt ccgtttatct caaactttag cattttggga ataaatgata tttgctatgc 181 tggttaaatt agattttagt taaatttcct gctgaagctc tagtacgata agcaacttga 241 cctaagtgta aagttgagac ttccttcagg tttatatagc ttgtgcgccg cttgggtacc 301 tcgggtcttc gagaagacct gttttagagc tagaaatagc aagttaaaat aaggctagtc 361 cgttatcaac ttgaaaaagt ggcaccgagt cggtgctttt tttctcggac tgatgacctg 421 agacgccgcg gtggagctcc agcttttgtt ccctttagtg agggttaatt gcgcgcttgg 481 cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca 541 acatacgagc cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca 601 cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc 661 attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt 721 cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact 781 caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag 841 caaaaggcca gcaaaaggcc aggaaccg //