LOCUS Exported 3174 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Module C, Promoter: TaU6, Gene: Esp3I ccdb cassette for single gRNA spacer cloning (+ BaeI site for GT donor cloning), Terminator: Pol III. ACCESSION . VERSION . KEYWORDS pMOD_C2518 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 3174) AUTHORS Cermak T, Curtin SJ, Gil-Humanes J, Cegan R, Kono TJY, Konecna E, Belanto JJ, Starker CG, Mathre JW, Greenstein RL, Voytas DF TITLE A multi-purpose toolkit to enable advanced genome engineering in plants. JOURNAL Plant Cell. 2017 May 18. pii: tpc.00922.2016. doi: 10.1105/tpc.16.00922. PUBMED 28522548 REFERENCE 2 (bases 1 to 3174) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..3174 /organism="synthetic DNA construct" /mol_type="other DNA" rep_origin 80..668 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 569..588 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 822..839 /label=L4440 /note="L4440 vector, forward primer" CDS complement(1240..1545) /codon_start=1 /gene="ccdB" /product="CcdB, a bacterial toxin that poisons DNA gyrase" /label=ccdB /note="Plasmids containing the ccdB gene cannot be propagated in standard E. coli strains." /translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK VSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI" primer_bind complement(1342..1361) /label=ccdB-fwd /note="ccdB gene, forward primer" misc_RNA 1872..1947 /label=gRNA scaffold /note="guide RNA scaffold for the Streptococcus pyogenes CRISPR/Cas9 system" promoter 2095..2186 /gene="bla" /label=AmpR promoter ORIGIN 1 gccttgacgg caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag 61 aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa 121 caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta ctaactcttt 181 ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc 241 cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa 301 tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa 361 gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg tgctcacagc 421 ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa 481 gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa 541 caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg 601 ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc 661 tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg 721 ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg 781 agtgagctga taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg 841 aagcggaaga gcgcccacct gcaacgccgg gaccaagccc gttattctga cagttctggt 901 gctcaacaca tttatattta tcaaggagca cattgttact cactgctagg agggaatcga 961 actaggaata ttgatcagag gaactacgag agagctgaag ataactgccc tctagctctc 1021 actgatctgg gtcgcatagt gagatgcagc ccacgtgagt tcagcaacgg tctagcgctg 1081 ggcttttagg cccgcatgat cgggcttttg tcgggtggtc gacgtgttca cgattgggga 1141 gagcaacgca gcagttcctc ttagtttagt cccacctcgc ctgtccagca gagttctgac 1201 cggtttataa actcgcttgc tgcatcagac ttggagacgt tatattcccc agaacatcag 1261 gttaatggcg tttttgatgt cattttcgcg gtggctgaga tcagccactt cttccccgat 1321 aacggaaacc ggcacactgg ccatatcggt ggtcatcatg cgccagcttt catccccgat 1381 atgcaccacc gggtaaagtt cacgggagac tttatctgac agcagacgtg cactggccag 1441 ggggatcacc atccgtcgcc cgggcgtgtc aataatatca ctctgtacat ccacaaacag 1501 acgataacgg ctctctcttt tataggtgta aaccttaaac tgcatttcac cagcccctgt 1561 tctcgtcagc aaaagagccg ttcatttcaa taaaccgggc gacctcagcc atcccttcct 1621 gattttccgc tttccagcgt tcggcacgca gacgacgggc ttcattctgc atggttgtgc 1681 ttaccagacc ggagatattg acatcatata tgccttgagc aactgatagc tgtcgctgtc 1741 aactgtcact gtaatacgct gcttcatagc atacctcctt ttgacatact tcgggtatac 1801 atatcagtat atattcttat accgcaaaaa tcagcgcgca aatacgcata ctgttatctg 1861 gcttcgtctc agttttagag ctagaaatag caagttaaaa taaggctagt ccgttatcaa 1921 cttgaaaaag tggcaccgag tcggtgcttt ttttcgcgta gtcctcggta atatcgcaga 1981 acaaaagtac ctgatatcga gtgtacttca agtcacaccg gcgagtgact agcaggtgat 2041 gaaaagcctt agggtcaggt ggcacttttc ggggaaatgt gcgatgaacc cctatttgtt 2101 tatttttcta aatacattca aatatgtatc cgctaatgag acaataaccc tgataaatgc 2161 ttcaattata ttgaaaaagg aagagtatga gtatccaaca tttccgtgtc gcccttattc 2221 ccttttttgc ggccttctgc cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa 2281 aagatgctga agatcagttg ggtgcccgag tgggttacat tgaactggat ctcaacagcg 2341 gcaaaatctt agagagtttt cgccccgaag aacgcttccc aatgatgagc actttcaaag 2401 ttctgctatg tggcgcggtg ttatcccgta ttgatgctgg gcaagagcaa ctcggtcgcc 2461 gcatacacta ttctcagaat gacttggttg agtattcacc agtcacagaa aagcatctta 2521 cggatggcat gacagtaaga gaattatgta gtgctgccat aaccatgagt gataacactg 2581 cggccaactt acttctgaca acgattggag gaccgaagga gctaaccgct tttttgcaca 2641 acatggggga tcatgtaact cgccttgatc gttgggaacc ggagctgaat gaagccatac 2701 caaacgacga gcgtgacacc acgatgcctg tagcaatggc aacaactctc cgcaaactat 2761 taactggcga actacttact cttgcttcac gccagcaact cattgactgg atggaggcgg 2821 acaaagttgc aggaccactt ctgcgctcgg cacttccggc tggctggttt attgctgata 2881 aatctggagc cggtgagcgt gggtctcgcg gtatcattgc agcactgggg ccagatggta 2941 agccctcccg tatcgtagtt atctacacga cggggagtca ggcaactatg gatgaacgaa 3001 atagacagat cgctgagata ggtgcctcac tgattaagca ttggtaactg tcagacgaag 3061 ggaaataaat agtagcccgc ctgatgtgcg ggcttttttt tggtggtggt taccagtgtc 3121 gcttagctac cgtaggtcct cttaggcggt ccgattagcg acatagtcat caat //