LOCUS Exported 1040 bp ds-DNA linear SYN 23-MAY-2021 DEFINITION tRNA and scaffold for the assembly of GBoligomers for the first position (positon [D1_n-1]) of a polycistronic tRNA-gRNA regulated by the U6-26 or U6-1 promoter (2-part multiplexing). ACCESSION . VERSION . KEYWORDS tRNA-gRNA position [D1_n-1] (GB1208) SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 1040) AUTHORS Vazquez-Vilar M, Bernabe-Orts JM, Fernandez-Del-Carmen A, Ziarsolo P, Blanca J, Granell A, Orzaez D TITLE A modular toolbox for gRNA-Cas9 genome engineering in plants based on the GoldenBraid standard. JOURNAL Plant Methods. 2016 Feb 1;12:10. doi: 10.1186/s13007-016-0101-2. eCollection 2016. PUBMED 26839579 REFERENCE 2 (bases 1 to 1040) AUTHORS . TITLE Direct Submission JOURNAL Exported May 23, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..1040 /organism="synthetic DNA construct" /mol_type="other DNA" regulatory 4..13 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" regulatory 8..17 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" promoter complement(316..334) /label=SP6 promoter /note="promoter for bacteriophage SP6 RNA polymerase" primer_bind complement(317..334) /label=SP6 /note="SP6 promoter, forward primer" primer_bind complement(352..368) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(352..368) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(365..387) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 376..392 /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)." promoter complement(400..430) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 445..466 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(583..600) /label=L4440 /note="L4440 vector, forward primer" primer_bind complement(834..853) /label=pBR322ori-F /note="pBR322 origin, forward primer" ORIGIN 1 ccggccgcca tggcggccgc gggaattcga ttcccgtctc gctcattgcc gactcggtgc 61 cactttttca agttgataac ggactagcct tattttaact tgctatttct agctctaaaa 121 ctgagacggc gcgcgccgtc tctgcaccag ccgggaatcg aacccgggtc tgtaccgtgg 181 cagggtacta ttctaccact agaccactgg tgctttgttc aatcgagcga gacgggatca 241 ctagtgaatt cgcggccgcc tgcaggtcga ccatatggga gagctcccaa cgcgttggat 301 gcatagcttg agtattctat agtgtcacct aaatagcttg gcgtaatcat ggtcatagct 361 gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat 421 aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc 481 actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg 541 cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct 601 gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt 661 atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc 721 caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga 781 gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata 841 ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac 901 cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg 961 taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc 1021 cgttcagccc gaccgctgcg //