LOCUS Exported 2707 bp ds-DNA circular SYN 19-JAN-2022 DEFINITION A crRNA expression plasmid for targeting a specific sequence.. ACCESSION . VERSION . KEYWORDS pCRISPR SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 2707) 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 2707) AUTHORS . TITLE Direct Submission JOURNAL Exported Jan 19, 2022 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..2707 /organism="synthetic DNA construct" /mol_type="other DNA" primer_bind complement(15..33) /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" CDS 212..1006 /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(266..285) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 876..895 /label=Neo-F /note="Neomycin resistance gene, forward primer" terminator 1037..1131 /label=lambda t0 terminator /note="transcription terminator from phage lambda" rep_origin 1219..1807 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 1708..1727 /label=pBR322ori-F /note="pBR322 origin, forward primer" terminator 1972..2058 /gene="Escherichia coli rrnB" /label=rrnB T1 terminator /note="transcription terminator T1 from the E. coli rrnB gene" misc_feature 2088..2219 /label=crRNA leader /note="crRNA leader sequence for the Streptococcus pyogenes CRISPR/Cas system" repeat_region 2220..2255 /label=DR /note="direct repeat for the Streptococcus pyogenes CRISPR/Cas system" repeat_region 2286..2321 /label=DR /note="direct repeat for the Streptococcus pyogenes CRISPR/Cas system" misc_feature 2362..2493 /label=crRNA leader /note="crRNA leader sequence for the Streptococcus pyogenes CRISPR/Cas system" repeat_region 2494..2529 /label=DR /note="direct repeat for the Streptococcus pyogenes CRISPR/Cas system" repeat_region 2560..2595 /label=DR /note="direct repeat for the Streptococcus pyogenes CRISPR/Cas system" promoter complement(2628..2701) /label=PLtetO-1 promoter /note="modified phage lambda PL promoter with tet operator sites (Lutz and Bujard, 1997)" protein_bind 2658..2676 /gene="tetO" /label=tet operator /bound_moiety="tetracycline repressor TetR" /note="bacterial operator O2 for the tetR and tetA genes" protein_bind 2683..2701 /gene="tetO" /label=tet operator /bound_moiety="tetracycline repressor TetR" /note="bacterial operator O2 for the tetR and tetA genes" ORIGIN 1 gtgaagacga aagggcctcg tgatacgcct atttttatag gttaatgtca tgataataat 61 ggtttcttag acgtcggaat tgccagctgg ggcgccctct ggtaaggttg ggaagccctg 121 caaagtaaac tggatggctt tcttgccgcc aaggatctga tggcgcaggg gatcaagatc 181 tgatcaagag acaggatgag gatcgtttcg catgattgaa caagatggat tgcacgcagg 241 ttctccggcc gcttgggtgg agaggctatt cggctatgac tgggcacaac agacaatcgg 301 ctgctctgat gccgccgtgt tccggctgtc agcgcagggg cgcccggttc tttttgtcaa 361 gaccgacctg tccggtgccc tgaatgaact gcaggacgag gcagcgcggc tatcgtggct 421 ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga 481 ctggctgcta ttgggcgaag tgccggggca ggatctcctg tcatctcacc ttgctcctgc 541 cgagaaagta tccatcatgg ctgatgcaat gcggcggctg catacgcttg atccggctac 601 ctgcccattc gaccaccaag cgaaacatcg catcgagcga gcacgtactc ggatggaagc 661 cggtcttgtc gatcaggatg atctggacga agagcatcag gggctcgcgc cagccgaact 721 gttcgccagg ctcaaggcgc gcatgcccga cggcgaggat ctcgtcgtga cccatggcga 781 tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt tctggattca tcgactgtgg 841 ccggctgggt gtggcggacc gctatcagga catagcgttg gctacccgtg atattgctga 901 agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga 961 ttcgcagcgc atcgccttct atcgccttct tgacgagttc ttctgagcgg gactctgggg 1021 ttcgagagct cgcttggact cctgttgata gatccagtaa tgacctcaga actccatctg 1081 gatttgttca gaacgctcgg ttgccgccgg gcgtttttta ttggtgagaa tccaagcact 1141 agtcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga 1201 aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac 1261 aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt 1321 tccgaaggta actggcttca gcagagcgca gataccaaat actgtccttc tagtgtagcc 1381 gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat 1441 cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag 1501 acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc 1561 cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag 1621 cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac 1681 aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg 1741 gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct 1801 atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc 1861 tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgcctttga 1921 gtgagctgat accgctcgcc gcagccgaac gccctaggtc tagggcggcg gatttgtcct 1981 actcaggaga gcgttcaccg acaaacaaca gataaaacga aaggcccagt ctttcgactg 2041 agcctttcgt tttatttgat gcctctagca cgcgtaccat gggatcctat ttcttaataa 2101 ctaaaaatat ggtataatac tcttaataaa tgcagtaata caggggcttt tcaagactga 2161 agtctagctg agacaaatag tgcgattacg aaatttttta gacaaaaata gtctacgagg 2221 ttttagagct atgctgtttt gaatggtccc aaaactgaga ccagtctcgg aagctcaaag 2281 gtctcgtttt agagctatgc tgttttgaat ggtcccaaaa cttcagcaca ctgagacttg 2341 ttgagttgaa ttcatggatc ctatttctta ataactaaaa atatggtata atactcttaa 2401 taaatgcagt aatacagggg cttttcaaga ctgaagtcta gctgagacaa atagtgcgat 2461 tacgaaattt tttagacaaa aatagtctac gaggttttag agctatgctg ttttgaatgg 2521 tcccaaaact gagaccagtc tcggaagctc aaaggtctcg ttttagagct atgctgtttt 2581 gaatggtccc aaaacttcag cacactgaga cttgttgagt tgaattcggt cagtgcgtcc 2641 tgctgatgtg ctcagtatct ctatcactga tagggatgtc aatctctatc actgataggg 2701 actcgag //