LOCUS Exported 2484 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION gRNA for cleavage at unc-109(n499) locus in C elegans. ACCESSION . VERSION . KEYWORDS pJA59 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 2484) AUTHORS Arribere JA, Bell RT, Fu BX, Artiles KL, Hartman PS, Fire AZ TITLE Efficient Marker-Free Recovery of Custom Genetic Modifications with CRISPR/Cas9 in Caenorhabditis elegans. JOURNAL Genetics. 2014 Aug 26. pii: genetics.114.169730. PUBMED 25161212 REFERENCE 2 (bases 1 to 2484) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..2484 /organism="synthetic DNA construct" /mol_type="other DNA" primer_bind 5..22 /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind 6..22 /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" promoter 197..546 /label=CeU6 promoter /note="promoter for the C. elegans R07E5.16 U6 snRNA gene (Dickinson et al., 2013)" misc_RNA 567..642 /label=gRNA scaffold /note="guide RNA scaffold for the Streptococcus pyogenes CRISPR/Cas9 system" primer_bind complement(785..801) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(785..801) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" terminator 849..880 /label=tonB terminator /note="bidirectional E. coli tonB-P14 transcription terminator" promoter 881..983 /label=cat promoter /note="promoter of the E. coli cat gene encoding chloramphenicol acetyltransferase" CDS 984..1799 /codon_start=1 /gene="aph(3')-Ia" /product="aminoglycoside phosphotransferase" /label=KanR /note="confers resistance to kanamycin in bacteria or G418 (Geneticin(R)) in eukaryotes" /translation="MSHIQRETSCLKPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNGDRVFRLAQAQSRMNNGLVGA SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF" primer_bind complement(1057..1076) /label=Kan-R /note="Kanamycin resistance gene, reverse primer" terminator 1823..1850 /label=T7Te terminator /note="phage T7 early transcription terminator" rep_origin complement(1862..2449) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(1942..1961) /label=pBR322ori-F /note="pBR322 origin, forward primer" ORIGIN 1 cccgtgtaaa acgacggcca gtttatctag tcagcttgat tctagctgat cgtggaccgg 61 aaggtgagcc agtgagttga ttgcagtcca gttacgctgg agtctgaggc tcgtcctgaa 121 tgatatgcga ccgccggagg gttgcgtttg agacgggcga cagatccagt cgcgctgctc 181 tcgtcgatcc aagcttcaaa aaaaactagc aataaaggaa taaaaaactg tacaccttaa 241 aggcgcacac tctgttttgc aaattttatt tttagttgtg aattttctgc tgagacttga 301 aaatagcaac tttagtacta ctataatttg tcaacctttt caaaaaaagc atgcaatttt 361 tgagaaactc ttataaaagc tattattaaa aaaacacctt ttttccaaaa ttattccaca 421 aaaaatatgt tatgaaatgc ctacaccctc tcacacacac tctttatact actctgtcaa 481 actcacgaga tgtctgccgc ctcttgtgtt gcccctatat aaacacctcc tattgcgaga 541 tgtcttggaa ctcgtgtcaa aacaacgttt tagagctaga aatagcaagt taaaataagg 601 ctagtccgtt atcaacttga aaaagtggca ccgagtcggt gctttttgtg aaatttgcta 661 gcggtgcgag cggatcgagc agtgtcgatc actactggac cgcgagctgt gctgcgaccc 721 gtgatcttac ggcattatac gtatgatcgg tccacgatca gctagattat ctagtcagct 781 tgatgtcata gctgtttcct gaggctcaat actgaccatt taaatcatac ctgacctcca 841 tagcagaaag tcaaaagcct ccgaccggag gcttttgact tgatcggcac gtaagaggtt 901 ccaactttca ccataatgaa ataagatcac taccgggcgt attttttgag ttatcgagat 961 tttcaggagc taaggaagct aaaatgagcc atattcaacg ggaaacgtct tgcttgaagc 1021 cgcgattaaa ttccaacatg gatgctgatt tatatgggta taaatgggct cgcgataatg 1081 tcgggcaatc aggtgcgaca atctatcgat tgtatgggaa gcccgatgcg ccagagttgt 1141 ttctgaaaca tggcaaaggt agcgttgcca atgatgttac agatgagatg gtcaggctaa 1201 actggctgac ggaatttatg cctcttccga ccatcaagca ttttatccgt actcctgatg 1261 atgcatggtt actcaccact gcgatcccag ggaaaacagc attccaggta ttagaagaat 1321 atcctgattc aggtgaaaat attgttgatg cgctggcagt gttcctgcgc cggttgcatt 1381 cgattcctgt ttgtaattgt ccttttaacg gcgatcgcgt atttcgtctc gctcaggcgc 1441 aatcacgaat gaataacggt ttggttggtg cgagtgattt tgatgacgag cgtaatggct 1501 ggcctgttga acaagtctgg aaagaaatgc ataaactctt gccattctca ccggattcag 1561 tcgtcactca tggtgatttc tcacttgata accttatttt tgacgagggg aaattaatag 1621 gttgtattga tgttggacga gtcggaatcg cagaccgata ccaggatctt gccatcctat 1681 ggaactgcct cggtgagttt tctccttcat tacagaaacg gctttttcaa aaatatggta 1741 ttgataatcc tgatatgaat aaattgcagt ttcacttgat gctcgatgag tttttctaat 1801 gaggacctaa atgtaatcac ctggctcacc ttcgggtggg cctttctgcg ttgctggcgt 1861 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgatgctca agtcagaggt 1921 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc 1981 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa 2041 gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct 2101 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta 2161 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg 2221 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc 2281 ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta 2341 cctcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg 2401 tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt 2461 gattttctac cgaagaaagg ccca //