LOCUS Exported 1058 bp ds-DNA linear SYN 19-MAY-2021 DEFINITION optimized lenti sgRNA cloning backbone with MS2 loops at tetraloop and stemloop 2 and EF1a-puro resistance marker. Contains BsmBI sites for insertion of spacer sequences.. ACCESSION . VERSION . KEYWORDS lenti sgRNA(MS2)_puro optimized backbone SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 1058) AUTHORS Joung J, Konermann S, Gootenberg JS, Abudayyeh OO, Platt RJ, Brigham MD, Sanjana NE, Zhang F TITLE Genome-scale CRISPR-Cas9 knockout and transcriptional activation screening. JOURNAL Nat Protoc. 2017 Apr;12(4):828-863. doi: 10.1038/nprot.2017.016. Epub 2017 Mar 23. PUBMED 28333914 REFERENCE 2 (bases 1 to 1058) AUTHORS . TITLE Direct Submission JOURNAL Exported May 19, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..1058 /organism="synthetic DNA construct" /mol_type="other DNA" promoter 6..246 /label=U6 promoter /note="RNA polymerase III promoter for human U6 snRNA" primer_bind 6..26 /label=hU6-F /note="Human U6 promoter, forward primer" primer_bind 177..196 /label=LKO.1 5' /note="Human U6 promoter, forward primer" misc_RNA 292..310 /label=MS2 stem loop /note="stem loop that binds the bacteriophage MS2 coat protein" misc_RNA 362..380 /label=MS2 stem loop /note="stem loop that binds the bacteriophage MS2 coat protein" misc_feature 472..589 /label=cPPT/CTS /note="central polypurine tract and central termination sequence of HIV-1" promoter 747..958 /label=EF-1-alpha core promoter /note="core promoter for human elongation factor EF-1-alpha" ORIGIN 1 ccagagaggg cctatttccc atgattcctt catatttgca tatacgatac aaggctgtta 61 gagagataat tagaattaat ttgactgtaa acacaaagat attagtacaa aatacgtgac 121 gtagaaagta ataatttctt gggtagtttg cagttttaaa attatgtttt aaaatggact 181 atcatatgct taccgtaact tgaaagtatt tcgatttctt ggctttatat atcttgtgga 241 aaggacgaaa cacccgagac gggataccgt ctctgtttta gagctaggcc aacatgagga 301 tcacccatgt ctgcagggcc tagcaagtta aaataaggct agtccgttat caacttggcc 361 aacatgagga tcacccatgt ctgcagggcc aagtggcacc gagtcggtgc ttttttggat 421 ccaagcttgg cgtaactaga tcttgagaca aatggcagta ttcatccaca attttaaaag 481 aaaagggggg attggggggt acagtgcagg ggaaagaata gtagacataa tagcaacaga 541 catacaaact aaagaattac aaaaacaaat tacaaaaatt caaaattttc gggtttatta 601 cagggacagc agagatccac tttggcgccg gctcgagggg gcccgggtgc aaagatggat 661 aaagttttaa acagagagga atctttgcag ctaatggacc ttctaggtct tgaaaggagt 721 gggaattggc tccggtgccc gtcagtgggc agagcgcaca tcgcccacag tccccgagaa 781 gttgggggga ggggtcggca attgatccgg tgcctagaga aggtggcgcg gggtaaactg 841 ggaaagtgat gtcgtgtact ggctccgcct ttttcccgag ggtgggggag aaccgtatat 901 aagtgcagta gtcgccgtga acgttctttt tcgcaacggg tttgccgcca gaacacaggt 961 aagtgccgtg tgtggttccc gcgggcctgg cctctttacg ggttatggcc cttgcgtgcc 1021 ttgaattact tccacctggc tgcagtacgt gattcttg //