LOCUS Exported 959 bp ds-DNA linear SYN 21-MAY-2021 DEFINITION Expresses gRNA from hybrid human U6/T7 promoter for both cellular expression and in vitro transcription from same promoter (1 G at initiation). ACCESSION . VERSION . KEYWORDS pU6T7G SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 959) AUTHORS Romanienko PJ, Giacalone J, Ingenito J, Wang Y, Isaka M, Johnson T, You Y, Mark WH TITLE A Vector with a Single Promoter for In Vitro Transcription and Mammalian Cell Expression of CRISPR gRNAs. JOURNAL PLoS One. 2016 Feb 5;11(2):e0148362. doi: 10.1371/journal.pone.0148362. eCollection 2016. PUBMED 26849369 REFERENCE 2 (bases 1 to 959) AUTHORS . TITLE Direct Submission JOURNAL Exported May 21, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..959 /organism="synthetic DNA construct" /mol_type="other DNA" primer_bind 41..60 /label=pENTR-R /note="pENTR vectors, reverse primer" primer_bind complement(191..213) /label=pGEX 3' /note="pGEX vectors, reverse primer" primer_bind 313..332 /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind 526..548 /label=M13/pUC Forward /note="In lacZ gene" primer_bind 608..628 /label=hU6-F /note="Human U6 promoter, forward primer" primer_bind 779..798 /label=LKO.1 5' /note="Human U6 promoter, forward primer" primer_bind 840..859 /label=T7 /note="T7 promoter, forward primer" promoter 840..858 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" misc_RNA 879..954 /label=gRNA scaffold /note="guide RNA scaffold for the Streptococcus pyogenes CRISPR/Cas9 system" ORIGIN 1 tgttggaatt taatcgcggc ctcgacgttt cccgttgaat atggctcata acaccccttg 61 tattactgtt tatgtaagca gacagtttta ttgttcatga tgatatattt ttatcttgtg 121 caatgtaaca tcagagattt tgagacacgg gccagagctg catcgcgcgt ttcggtgatg 181 acggtgaaaa cctctgacac atgcagctcc cggagacggt cacagcttgt ctgtaagcgg 241 atgccgggag cagacaagcc cgtcagggcg cgtcagcggg tgttggcggg tgtcggggct 301 ggcttaacta tgcggcatca gagcagattg tactgagagt gcaccatatg cggtgtgaaa 361 taccgcacag atgcgtaagg agaaaatacc gcatcaggcg ccattcgcca ttcaggctgc 421 gcaactgttg ggaagggcga tcggtgcggg cctcttcgct attacgccag ctggcgaaag 481 ggggatgtgc tgcaaggcga ttaagttggg taacgccagg gttttcccag tcacgacgtt 541 gtaaaacgac ggccagagaa ttcgagctcg gtacctcgcg aatacatcta gaaaggtcgg 601 gcaggaagag ggcctatttc ccatgattcc ttcatatttg catatacgat acaaggctgt 661 tagagagata attagaatta atttgactgt aaacacaaag atattagtac aaaatacgtg 721 acgtagaaag taataatttc ttgggtagtt tgcagtttta aaattatgtt ttaaaatgga 781 ctatcatatg cttaccgtaa cttgaaagta tttcgatttc ttggctttat atatcttgct 841 aatacgactc actataggga gaccgagaga gggtctcagt tttagagcta gaaatagcaa 901 gttaaaataa ggctagtccg ttatcaactt gaaaaagtgg caccgagtcg gtgcttttt //