LOCUS Exported 1423 bp ds-DNA linear SYN 17-MAY-2021 DEFINITION Catalytically inactive Cas9 from S. pyogenes with P2A-EGFP under the EF1a core promoter, and cloning backbone for sgRNA. Contains BsmBI sites for insertion of spacer sequences.. ACCESSION . VERSION . KEYWORDS Lenti-(BB)-EF1a-KRAB-dCas9-P2A-EGFP SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 1423) TITLE Ferrer Lab Plasmids REFERENCE 2 (bases 1 to 1423) AUTHORS . TITLE Direct Submission JOURNAL Exported May 17, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..1423 /organism="synthetic DNA construct" /mol_type="other DNA" CDS 294..320 /codon_start=1 /product="HA (human influenza hemagglutinin) epitope tag" /label=HA /translation="YPYDVPDYA" CDS 339..359 /codon_start=1 /product="nuclear localization signal of SV40 (simian virus 40) large T antigen" /label=SV40 NLS /translation="PKKKRKV" CDS 366..386 /codon_start=1 /product="nuclear localization signal of SV40 (simian virus 40) large T antigen" /label=SV40 NLS /translation="PKKKRKV" CDS 429..482 /codon_start=1 /product="2A peptide from Thosea asigna virus capsid protein" /label=T2A /note="Eukaryotic ribosomes fail to insert a peptide bond between the Gly and Pro residues, yielding separate polypeptides." /translation="EGRGSLLTCGDVEENPGP" CDS 483..998 /codon_start=1 /product="N-terminal fragment of mVenus for use in bimolecular fluorescence complementation (BiFC) (Kodama and Hu, 2010)" /label=VN173 /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV NFKIRHNIE" primer_bind complement(525..546) /label=EGFP-N /note="EGFP, reverse primer" primer_bind complement(786..805) /label=EXFP-R /note="For distinguishing EGFP variants, reverse primer" ORIGIN 1 nnnnnntnnn gnntntnnag agagtgatcc tggccgacgc taatctggac aaggtgctga 61 gcgcctacaa caagcacaga gacaagccta tcagagagca ggccgagaat atcatccacc 121 tgtttaccct gaccagtctg ggagcccctg ccgccttcaa gtactttgac accaccatcg 181 accggaagag gtacaccagc accaaagagg tgctggacgc caccctgatc caccagagca 241 tcaccggcct gtacgagaca cggatcgacc tgtctcagct gggaggcgac gcctatccct 301 atgacgtgcc cgattatgcc agcctgggca gcggctcccc caagaaaaaa cgcaaggtgg 361 aagatcctaa gaaaaagcgg aaagtggacg gcattggtag tgggagcaac ggcagcagcg 421 gggatcccga gggcagagga agtctgctaa catgcggtga cgtcgaggag aatcctggcc 481 cagtgagcaa gggcgaggag ctgttcaccg gggtggtgcc catcctggtc gagctggacg 541 gcgacgtaaa cggccacaag ttcagcgtgt ccggcgaggg cgagggcgat gccacctacg 601 gcaagctgac cctgaagttc atctgcacca ccggcaagct gcccgtgccc tggcccaccc 661 tcgtgaccac cctgacctac ggcgtgcagt gcttcagccg ctaccccgac cacatgaagc 721 agcacgactt cttcaagtcc gccatgcccg aaggctacgt ccaggagcgc accatcttct 781 tcaaggacga cggcaactac aagacccgcg ccgaggtgaa gttcgagggc gacaccctgg 841 tgaaccgcat cgagctgaag ggcatcgact tcaaggagga cggcaacatc ctggggcaca 901 agctggagta caactacaac agccacaacg tctatatcat ggccgacaag cagaagaacg 961 gcatcaaggt gaacttcaag atccgccaca acatcgagga cggcagcgtg cagctcgccg 1021 accactacca gcagaacacc cccatcggcg acggccccgt gctgctgccc gacaaccact 1081 acctgagcac ccagtccgcc ctgaacaaag accccaacga gaagcgcgat cacatggtcc 1141 tggctggaat tcgtgaccgc cgcccgggat nnctcncggc atggacaanc tgtacaagga 1201 attctaancg nnttaannga caatcaaccn tnggnattnc aaaattgtgg aagaaatgan 1261 gggggattct taacannntn gcccntttta cccnagtggn aaacccngct tnanagccct 1321 tgtttaagcc nnntggttcc cccaaggggt tnttttttct ccccntgaaa aaacccggnt 1381 ncggncctct ttnnnnnaan nntgggcccc ttncntgnaa agg //