LOCUS Exported 2433 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION A crRNA expression plasmid specific to the rpsL allele.. ACCESSION . VERSION . KEYWORDS pCRISPR::rpsL SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 2433) AUTHORS Jiang W, Bikard D, Cox D, Zhang F, Marraffini LA TITLE RNA-guided editing of bacterial genomes using CRISPR-Cas systems. JOURNAL Nat Biotechnol. 2013 Jan 29. doi: 10.1038/nbt.2508. PUBMED 23360965 REFERENCE 2 (bases 1 to 2433) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..2433 /organism="synthetic DNA construct" /mol_type="other DNA" primer_bind complement(19..37) /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" CDS 216..1010 /codon_start=1 /gene="aph(3')-II (or nptII)" /product="aminoglycoside phosphotransferase from Tn5" /label=NeoR/KanR /note="confers resistance to neomycin, kanamycin, and G418 (Geneticin(R))" /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF" primer_bind complement(270..289) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 880..899 /label=Neo-F /note="Neomycin resistance gene, forward primer" terminator 1041..1135 /label=lambda t0 terminator /note="transcription terminator from phage lambda" rep_origin 1223..1811 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 1712..1731 /label=pBR322ori-F /note="pBR322 origin, forward primer" terminator 1976..2062 /gene="Escherichia coli rrnB" /label=rrnB T1 terminator /note="transcription terminator T1 from the E. coli rrnB gene" misc_feature 2092..2223 /label=crRNA leader /note="crRNA leader sequence for the Streptococcus pyogenes CRISPR/Cas system" repeat_region 2224..2259 /label=DR /note="direct repeat for the Streptococcus pyogenes CRISPR/Cas system" repeat_region 2290..2325 /label=DR /note="direct repeat for the Streptococcus pyogenes CRISPR/Cas system" promoter complement(2358..2431) /label=PLtetO-1 promoter /note="modified phage lambda PL promoter with tet operator sites (Lutz and Bujard, 1997)" protein_bind 2388..2406 /gene="tetO" /label=tet operator /bound_moiety="tetracycline repressor TetR" /note="bacterial operator O2 for the tetR and tetA genes" protein_bind 2413..2431 /gene="tetO" /label=tet operator /bound_moiety="tetracycline repressor TetR" /note="bacterial operator O2 for the tetR and tetA genes" ORIGIN 1 cgaggtgaag acgaaagggc ctcgtgatac gcctattttt ataggttaat gtcatgataa 61 taatggtttc ttagacgtcg gaattgccag ctggggcgcc ctctggtaag gttgggaagc 121 cctgcaaagt aaactggatg gctttcttgc cgccaaggat ctgatggcgc aggggatcaa 181 gatctgatca agagacagga tgaggatcgt ttcgcatgat tgaacaagat ggattgcacg 241 caggttctcc ggccgcttgg gtggagaggc tattcggcta tgactgggca caacagacaa 301 tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg gttctttttg 361 tcaagaccga cctgtccggt gccctgaatg aactgcagga cgaggcagcg cggctatcgt 421 ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact gaagcgggaa 481 gggactggct gctattgggc gaagtgccgg ggcaggatct cctgtcatct caccttgctc 541 ctgccgagaa agtatccatc atggctgatg caatgcggcg gctgcatacg cttgatccgg 601 ctacctgccc attcgaccac caagcgaaac atcgcatcga gcgagcacgt actcggatgg 661 aagccggtct tgtcgatcag gatgatctgg acgaagagca tcaggggctc gcgccagccg 721 aactgttcgc caggctcaag gcgcgcatgc ccgacggcga ggatctcgtc gtgacccatg 781 gcgatgcctg cttgccgaat atcatggtgg aaaatggccg cttttctgga ttcatcgact 841 gtggccggct gggtgtggcg gaccgctatc aggacatagc gttggctacc cgtgatattg 901 ctgaagagct tggcggcgaa tgggctgacc gcttcctcgt gctttacggt atcgccgctc 961 ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga gcgggactct 1021 ggggttcgag agctcgcttg gactcctgtt gatagatcca gtaatgacct cagaactcca 1081 tctggatttg ttcagaacgc tcggttgccg ccgggcgttt tttattggtg agaatccaag 1141 cactagtcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg 1201 tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc 1261 aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc 1321 tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtc cttctagtgt 1381 agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc 1441 taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact 1501 caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac 1561 agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag 1621 aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg 1681 gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg 1741 tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga 1801 gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt 1861 ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt attaccgcct 1921 ttgagtgagc tgataccgct cgccgcagcc gaacgcccta ggtctagggc ggcggatttg 1981 tcctactcag gagagcgttc accgacaaac aacagataaa acgaaaggcc cagtctttcg 2041 actgagcctt tcgttttatt tgatgcctct agcacgcgta ccatgggatc ctatttctta 2101 ataactaaaa atatggtata atactcttaa taaatgcagt aatacagggg cttttcaaga 2161 ctgaagtcta gctgagacaa atagtgcgat tacgaaattt tttagacaaa aatagtctac 2221 gaggttttag agctatgctg ttttgaatgg tcccaaaact actttacgca gcgcggagtt 2281 cggttttttg ttttagagct atgctgtttt gaatggtccc aaaacttcag cacactgaga 2341 cttgttgagt tgaattcggt cagtgcgtcc tgctgatgtg ctcagtatct ctatcactga 2401 tagggatgtc aatctctatc actgataggg act //