LOCUS Exported 917 bp ds-DNA linear SYN 20-MAY-2021 DEFINITION Lentiviral CRISPR-Cas9 delivery for SpCas9 and sgRNA. Coexpresses tagRFP via P2A cleavage site. EFS Promoter driven. ACCESSION . VERSION . KEYWORDS pL-CRISPR.EFS.tRFP SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 917) AUTHORS Heckl D, Kowalczyk MS, Yudovich D, Belizaire R, Puram RV, McConkey ME, Thielke A, Aster JC, Regev A, Ebert BL TITLE Generation of mouse models of myeloid malignancy with combinatorial genetic lesions using CRISPR-Cas9 genome editing. JOURNAL Nat Biotechnol. 2014 Jun 22. doi: 10.1038/nbt.2951. PUBMED 24952903 REFERENCE 2 (bases 1 to 917) AUTHORS . TITLE Direct Submission JOURNAL Exported May 20, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..917 /organism="synthetic DNA construct" /mol_type="other DNA" CDS complement(41..754) /codon_start=1 /product="monomeric derivative of red fluorescent protein from Entacmaea quadricolor (Merzlyak et al., 2007)" /label=TagRFP /note="mammalian codon-optimized" /translation="MVSKGEELIKENMHMKLYMEGTVNNHHFKCTSEGEGKPYEGTQTM RIKVVEGGPLPFAFDILATSFMYGSRTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGG VLTATQDTSLQDGCLIYNVKIRGVNFPSNGPVMQKKTLGWEANTEMLYPADGGLEGRSD MALKLVGGGHLICNFKTTYRSKKPAKNLKMPGVYYVDHRLERIKEADKETYVEQHEVAV ARYCDLPSKLGHKLN" CDS complement(755..811) /codon_start=1 /product="2A peptide from porcine teschovirus-1 polyprotein" /label=P2A /note="Eukaryotic ribosomes fail to insert a peptide bond between the Gly and Pro residues, yielding separate polypeptides." /translation="ATNFSLLKQAGDVEENPGP" CDS complement(827..874) /codon_start=1 /product="bipartite nuclear localization signal from nucleoplasmin" /label=nucleoplasmin NLS /translation="KRPAATKKAGQAKKKK" ORIGIN 1 aaattttgta atccagaggt tgattgtcga cttaacgcgt tcaattaagt ttgtgcccca 61 gtttgctagg gaggtcgcag tatctggcca cagccacctc gtgctgctcg acgtaggtct 121 ctttgtcggc ctccttgatt ctttccagtc tgtggtccac atagtagacg ccgggcatct 181 tgaggttctt agcgggtttc ttggatctgt atgtggtctt gaagttgcag atcaggtggc 241 ccccgcccac gagcttcagg gccatgtcgc ttctgccttc caggccgccg tcagcggggt 301 acagcatctc ggtgttggcc tcccagccga gtgttttctt ctgcatcaca gggccgttgg 361 atgggaagtt cacccctctg atcttgacgt tgtagatgag gcagccgtcc tggaggctgg 421 tgtcctgggt agcggtcagc acgcccccgt cttcgtatgt ggtgactctc tcccatgtga 481 agccctcagg gaaggactgc ttaaagaagt cggggatgcc ctgggtgtgg ttgatgaagg 541 ttctgctgcc gtacatgaag ctggtagcca ggatgtcgaa ggcgaagggg agagggccgc 601 cctcgaccac cttgattctc atggtctggg tgccctcgta gggcttgcct tcgccctcgg 661 atgtgcactt gaagtggtgg ttgttcacgg tgccctccat atacagcttc atgtgcatgt 721 tctccttaat cagctcttcg cccttagaca ccatggggcc ggggttctcc tccacgtcgc 781 cggcctgctt cagcaggctg aagttggtgg cgccgctgcc gctagctttc tttttcttag 841 cttgaccagc tttcttagta gcagcaggac gcttgtcgcc tcccagctga gacaggtcga 901 tccgtgtctc gtacagg //