LOCUS Exported 803 bp ds-DNA linear SYN 17-MAY-2021 DEFINITION Mammalian U6-driven sgRNA (SPm) targeting GTCCCCTCCACCCCACAGTG. ACCESSION . VERSION . KEYWORDS M-SP-sgRNA SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 803) AUTHORS Esvelt KM, Mali P, Braff JL, Moosburner M, Yaung SJ, Church GM TITLE Orthogonal Cas9 proteins for RNA-guided gene regulation and editing. JOURNAL Nat Methods. 2013 Sep 29. doi: 10.1038/nmeth.2681. PUBMED 24076762 REFERENCE 2 (bases 1 to 803) AUTHORS . TITLE Direct Submission JOURNAL Exported May 17, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..803 /organism="synthetic DNA construct" /mol_type="other DNA" protein_bind 92..116 /gene="mutant version of attB" /label=attB2 /bound_moiety="BP Clonase(TM)" /note="recombination site for the Gateway(R) BP reaction" misc_RNA complement(128..203) /label=gRNA scaffold /note="guide RNA scaffold for the Streptococcus pyogenes CRISPR/Cas9 system" promoter complement(232..472) /label=U6 promoter /note="RNA polymerase III promoter for human U6 snRNA" primer_bind complement(282..301) /label=LKO.1 5' /note="Human U6 promoter, forward primer" primer_bind complement(452..472) /label=hU6-F /note="Human U6 promoter, forward primer" primer_bind complement(611..630) /label=T7 /note="T7 promoter, forward primer" promoter complement(612..630) /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" primer_bind complement(637..654) /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind complement(637..653) /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" primer_bind complement(646..668) /label=M13/pUC Forward /note="In lacZ gene" ORIGIN 1 ctatagaata ctcaagctat gcatcaagct tggtaccgag ctcggatcca ctagtaacgg 61 ccgccagtgt gctggaattc gccctttaat gccaactttg tacaagaaag ctgggtctag 121 aaaaaaagca ccgactcggt gccacttttt caagttgata acggactagc cttattttaa 181 cttgctattt ctagctctaa aaccactgtg gggtggaggg gacggtgttt cgtcctttcc 241 acaagatata taaagccaag aaatcgaaat actttcaagt tacggtaagc atatgatagt 301 ccattttaaa acataatttt aaaactgcaa actacccaag aaattattac tttctacgtc 361 acgtattttg tactaatatc tttgtgttta cagtcaaatt aattctaatt atctctctaa 421 cagccttgta tcgtatatgc aaatatgaag gaatcatggg aaataggccc tcttcctgcc 481 cgaccttggt accggatcca gtcgactgaa ttggttcctt taaagcctgc ttttttgtac 541 aaagggcgaa ttctgcagat atccatcaca ctggcggccg ctcgagcatg catctagagg 601 gcccaattcg ccctatagtg agtcgtatta caattcactg gccgtcgttt tacaacgtcg 661 tgactgggaa aaccctggcg ttacccaact taatcgcctt gcagcacatc cccctttcgc 721 cagctggcgt aatagcgaag aggcccgcac cgatcgccct tcccaacagt tgcgcagcct 781 atacgtacgg cagtttaagg ttt //