LOCUS Exported 2960 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION to drive the sgRNA expression under a U6 promoter. ACCESSION . VERSION . KEYWORDS sgRNA with U6 promoter SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 2960) AUTHORS Chen C, Fenk LA, de Bono M TITLE Efficient genome editing in Caenorhabditis elegans by CRISPR-targeted homologous recombination. JOURNAL Nucleic Acids Res. 2013 Sep 5. PUBMED 24013562 REFERENCE 2 (bases 1 to 2960) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..2960 /organism="synthetic DNA construct" /mol_type="other DNA" primer_bind 278..300 /label=M13/pUC Forward /note="In lacZ gene" primer_bind 292..309 /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind 293..309 /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" primer_bind 320..339 /label=T7 /note="T7 promoter, forward primer" promoter 320..338 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" misc_RNA 699..774 /label=gRNA scaffold /note="guide RNA scaffold for the Streptococcus pyogenes CRISPR/Cas9 system" primer_bind complement(1103..1120) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(1213..1801) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(1293..1312) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(1972..2832) /codon_start=1 /gene="bla" /product="beta-lactamase" /label=AmpR /note="confers resistance to ampicillin, carbenicillin, and related antibiotics" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS LIKHW" primer_bind 2595..2614 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(2833..2937) /gene="bla" /label=AmpR promoter ORIGIN 1 ctaaattgta agcgttaata ttttgttaaa attcgcgtta aatttttgtt aaatcagctc 61 attttttaac caataggccg aaatcggcaa aatcccttat aaatcaaaag aatagaccga 121 gatagggttg agtggccgct acagggcgct cccattcgcc attcaggctg cgcaactgtt 181 gggaagggcg tttcggtgcg ggcctcttcg ctattacgcc agctggcgaa agggggatgt 241 gctgcaaggc gattaagttg ggtaacgcca gggttttccc agtcacgacg ttgtaaaacg 301 acggccagtg agcgcgacgt aatacgactc actatagggc gaattggcgg aaggccgtca 361 aggccacgtg tcttgtccag gcgcgccacc caaatgtttt tataatttgg caaattacca 421 aaaatgtgac tggaagcatt aacaaaattt tgaacaatat tcgaggtttc aaaatcatga 481 tattttagat attttggact gttttgagtc aaattgttaa tgacagcatt ttaagagctc 541 ccaacacata gtgtttccaa tgttataccc aatcaataat agcaagtcaa taaactacct 601 ctacactatt tctggtaata ggcgaaacct ctacactgtc agtcactttg taaggtgtgc 661 ctatgtattt ccataatatt tcatacaaat tgaagcttgt tttagagcta gaaatagcaa 721 gttaaaataa ggctagtccg ttatcaactt gaaaaagtgg caccgagtcg gtgctttttt 781 tgactaattt ctggtattat tgtttagaat tgtaagaaaa ttttacattc cagaaaatgt 841 atccaaaaaa cttacgcaaa ctcggagcgt atagttgcat gaagtgctgg atccgaaata 901 cgaacgacaa attgttgata ctttgctttc aaactctcgt aataaggcat cacaaagttc 961 tcaattttat taattaatgg agcacaagac tggcctcatg ggccttccgc tcactgcccg 1021 ctttccagtc gggaaacctg tcgtgccagc tgcattaaca tggtcatagc tgtttccttg 1081 cgtattgggc gctctccgct tcctcgctca ctgactcgct gcgctcggtc gttcgggtaa 1141 agcctggggt gcctaatgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc 1201 gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc 1261 aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag 1321 ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct 1381 cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta 1441 ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc 1501 cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc 1561 agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt 1621 gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct 1681 gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc 1741 tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca 1801 agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta 1861 agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa 1921 atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg 1981 cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg 2041 actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc 2101 aatgataccg cgagaaccac gctcaccggc tccagattta tcagcaataa accagccagc 2161 cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa 2221 ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc 2281 cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg 2341 ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc 2401 cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat 2461 ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg 2521 tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc 2581 ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg 2641 aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat 2701 gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg 2761 gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg 2821 ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct 2881 catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac 2941 atttccccga aaagtgccac //