LOCUS Exported 884 bp ds-DNA linear SYN 18-MAY-2021 DEFINITION U6 driven sgRNA cloning vector where guide sequences are inserted between BfuAI sites. ACCESSION . VERSION . KEYWORDS pLKO.1-puro U6 sgRNA BfuAI stuffer SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 884) AUTHORS Kearns NA, Genga RM, Enuameh MS, Garber M, Wolfe SA, Maehr R TITLE Cas9 effector-mediated regulation of transcription and differentiation in human pluripotent stem cells. JOURNAL Development. 2014 Jan;141(1):219-23. doi: 10.1242/dev.103341. PUBMED 24346702 REFERENCE 2 (bases 1 to 884) AUTHORS . TITLE Direct Submission JOURNAL Exported May 18, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..884 /organism="synthetic DNA construct" /mol_type="other DNA" primer_bind 125..144 /label=LKO.1 5' /note="Human U6 promoter, forward primer" misc_RNA 236..311 /label=gRNA scaffold /note="guide RNA scaffold for the Streptococcus pyogenes CRISPR/Cas9 system" misc_feature 360..477 /label=cPPT/CTS /note="central polypurine tract and central termination sequence of HIV-1" ORIGIN 1 ggctgttaga gagataattg gaattaattt gactgtaaac acaaagatat tagtacaaaa 61 tacgtgacgt agaaagtaat aatttcttgg gtagtttgca gttttaaaat tatgttttaa 121 aatggactat catatgctta ccgtaacttg aaagtatttc gatttcttgg ctttatatat 181 cttgtggaaa ggacgaaaca ccggtgtgca ggtgagtgat caagaacctg ctgacgtttt 241 agagctagaa atagcaagtt aaaataaggc tagtccgtta tcaacttgaa aaagtggcac 301 cgagtcggtg ctttttttga attctcgacc tcgagacaaa tggcagtatt catccacaat 361 tttaaaagaa aaggggggat tggggggtac agtgcagggg aaagaatagt agacataata 421 gcaacagaca tacaaactaa agaattacaa aaacaaatta caaaaattca aaattttcgg 481 gtttattaca gggacagcag agatccactt tggccgcggc tcgagggggt tggggttgcg 541 ccttttccaa ggcagccctg ggtttgcgca gggacgcggc tgctctgggc gtggttccgg 601 gaaacgcagc ggcgccgacc ctgggtctcg cacattcttc acgtccgttc gcagcgtcac 661 ccggatcttc gccgctaccc ttgtgggccc cccggcgacg cttcctgctc cgcccctaag 721 tcgggaaggt tccttgcggt tcgcggcgtg ccggacgtga caaacggaag ccgcacgtct 781 cactagtacc ctcgcagacg gacagcgcca gggagcaatg gcagcgcgcc gaccgcgatg 841 ggctgtggcc aatagcggct gctcagcagg gcgcgccgag agca //