LOCUS Exported 999 bp ds-DNA linear SYN 20-MAY-2021 DEFINITION for e. coli expression, Golden Gate Compatible TALEN incorporated with C Terminal 6xHIS Tag, Codon Optimized FokI. ACCESSION . VERSION . KEYWORDS pET-Sangamo-HIS SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 999) TITLE Bao Lab Golden Gate plasmids JOURNAL Unpublished REFERENCE 2 (bases 1 to 999) AUTHORS . TITLE Direct Submission JOURNAL Exported May 20, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..999 /organism="synthetic DNA construct" /mol_type="other DNA" promoter complement(3..79) /gene="lacI" /label=lacI promoter primer_bind 285..304 /label=pBRrevBam /note="pBR322 vectors, tet region, downstream of BamHI, reverse primer" primer_bind 388..407 /label=T7 /note="T7 promoter, forward primer" promoter 388..406 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" protein_bind 407..431 /label=lac operator /bound_moiety="lac repressor encoded by lacI" /note="The lac repressor binds to the lac operator to inhibit transcription in E. coli. This inhibition can be relieved by adding lactose or isopropyl-beta-D-thiogalactopyranoside (IPTG)." RBS 446..468 /note="efficient ribosome binding site from bacteriophage T7 gene 10 (Olins and Rangwala, 1989)" CDS 481..546 /codon_start=1 /product="three tandem FLAG(R) epitope tags, followed by an enterokinase cleavage site" /label=3xFLAG /translation="DYKDHDGDYKDHDIDYKDDDDK" CDS 553..573 /codon_start=1 /product="nuclear localization signal of SV40 (simian virus 40) large T antigen" /label=SV40 NLS /translation="PKKKRKV" ORIGIN 1 cattcaccac cctgaattga ctctcttccg ggcgctatca tgccataccg cgaaaggttt 61 tgcgccattc gatggtgtcc gggatctcga cgctctccct tatgcgactc ctgcattagg 121 aagcagccca gtagtaggtt gaggccgttg agcaccgccg ccgcaaggaa tggtgcatgc 181 aaggagatgg cgcccaacag tcccccggcc acggggcctg ccaccatacc cacgccgaaa 241 caagcgctca tgagcccgaa gtggcgagcc cgatcttccc catcggtgat gtcggcgata 301 taggcgccag caaccgcacc tgtggcgccg gtgatgccgg ccacgatgcg tccggcgtag 361 aggatcgaga tctcgatccc gcgaaattaa tacgactcac tataggggaa ttgtgagcgg 421 ataacaattc ccctctagaa ataattttgt ttaactttaa gaaggagata taccatggcg 481 gactacaaag accatgacgg tgattataaa gatcatgaca tcgattacaa ggatgacgat 541 gacaagatgg cccccaagaa gaagaggaag gtgggcattc accgcggggt acctatggtg 601 gacttgagga cactcggtta ttcgcaacag caacaggaga aaatcaagcc taaggtcagg 661 agcaccgtcg cgcaacacca cgaggcgctt gtggggcatg gcttcactca tgcgcatatt 721 gtcgcgcttt cacagcaccc tgcggcgctt gggacggtgg ctgtcaaata ccaagatatg 781 attgcggccc tgcccgaagc cacgcacgag gcaattgtag gggtcggtaa acagtggtcg 841 ggagcgcgag cacttgaggc gctgctgact gtggcgggtg agcttagggg gcctccgctc 901 cagctcgaca ccgggcagct gctgaagatc gcgaagagag ggggagtaac agcggtagag 961 gcagtgcacg cctggcgcaa tgcgctcacc ggtgccccc //