LOCUS Exported 3286 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION Golden Gate Level 0 cassette encoding the U6 promoter. ACCESSION . VERSION . KEYWORDS pCR8/GW:U6 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 3286) AUTHORS Hopes A, Nekrasov V, Kamoun S, Mock T TITLE Editing of the urease gene by CRISPR-Cas in the diatom Thalassiosira pseudonana JOURNAL Plant Methods 2016, 12:49 REFERENCE 2 (bases 1 to 3286) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..3286 /organism="synthetic DNA construct" /mol_type="other DNA" rep_origin complement(63..651) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(143..162) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(744..1535) /codon_start=1 /gene="aadA" /product="aminoglycoside adenylyltransferase (Murphy, 1985)" /label=SmR /note="confers resistance to spectinomycin and streptomycin" /translation="MREAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK" primer_bind 1966..1982 /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind 1966..1982 /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind 1987..2006 /label=T7 /note="T7 promoter, forward primer" promoter 1987..2005 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" protein_bind 2023..2122 /gene="mutant version of attL" /label=attL2 /bound_moiety="LR Clonase(TM)" /note="recombination site for the Gateway(R) LR reaction" protein_bind complement(2619..2718) /gene="mutant version of attL" /label=attL1 /bound_moiety="LR Clonase(TM)" /note="recombination site for the Gateway(R) LR reaction" primer_bind complement(2734..2751) /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind complement(2734..2750) /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" primer_bind complement(2743..2765) /label=M13/pUC Forward /note="In lacZ gene" terminator 2814..2900 /gene="Escherichia coli rrnB" /label=rrnB T1 terminator /note="transcription terminator T1 from the E. coli rrnB gene" terminator 2992..3019 /label=rrnB T2 terminator /note="transcription terminator T2 from the E. coli rrnB gene" primer_bind complement(3178..3195) /label=L4440 /note="L4440 vector, forward primer" ORIGIN 1 aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg 61 tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg 121 tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg 181 cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga 241 agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc 301 tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt 361 aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact 421 ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg 481 cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt 541 accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt 601 ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct 661 ttgatctttt ctacggggtc tgacgctcag tggaacgacg cgtaactcac gttaagggat 721 tttggtcatg ggtggctcga gggttatttg ccgactacct tggtgatctc gcctttcacg 781 tagtggacaa attcttccaa ctgatctgcg cgcgaggcca agcgatcttc ttcttgtcca 841 agataagcct gtctagcttc aagtatgacg ggctgatact gggccggcag gcgctccatt 901 gcccagtcgg cagcgacatc cttcggcgcg attttgccgg ttactgcgct gtaccaaatg 961 cgggacaacg taagcactac atttcgctca tcgccagccc agtcgggcgg cgagttccat 1021 agcgttaagg tttcatttag cgcctcaaat agatcctgtt caggaaccgg atcaaagagt 1081 tcctccgccg ctggacctac caaggcaacg ctatgttctc ttgcttttgt cagcaagata 1141 gccagatcaa tgtcgatcgt ggctggctcg aagatacctg caagaatgtc attgcgctgc 1201 cattctccaa attgcagttc gcgcttagct ggataacgcc acggaatgat gtcgtcgtgc 1261 acaacaatgg tgacttctac agcgcggaga atctcgctct ctccagggga agccgaagtt 1321 tccaaaaggt cgttgatcaa agctcgccgc gttgtttcat caagccttac ggtcaccgta 1381 accagcaaat caatatcact gtgtggcttc aggccgccat ccactgcgga gccgtacaaa 1441 tgtacggcca gcaacgtcgg ttcgagatgg cgctcgatga cgccaactac ctctgatagt 1501 tgagtcgata cttcggcgat caccgcttcc ctcataatgt ttaactttgt tttagggcga 1561 ctgccctgct gcgtaacatc gttgctgctc cataacatca aacatcgacc cacggcgtaa 1621 cgcgcttgct gcttggatgc ccgaggcata gactgtaccc caaaaaaaca gtcataacaa 1681 gccatgaaaa ccgccactgc gccgttacca ccgctgcgtt cggtcaaggt tctggaccag 1741 ttgcgtgagc gcatacgcta cttgcattac agcttacgaa ccgaacaggc ttatgtccac 1801 tgggttcgtg ccttcatccg tttccacggt gtgcgtcacc cggcaacctt gggtagcagc 1861 gaagtcgagg catttctgtc ctggctggtc tagaggaggc atgatgatat atttttatct 1921 tgtgcaatgt aacatcagag attttgagac acgggccaga gctgccagga aacagctatg 1981 accatgtaat acgactcact ataggggata tcagctggat ggcaaataat gattttattt 2041 tgactgatag tgacctgttc gttgcaacaa attgatgagc aattattttt tataatgcca 2101 actttgtaca agaaagctgg gtcgaattcg cccttcggtc tcaggagctt catcaagaga 2161 gcaaccaaca aagaggaggc gaacacggtg cacagcaccg tctgcatact tccgatttgc 2221 aattcgcaac atatgataca tcatccatct ctctgagaaa actgggcact gttagaatct 2281 gggatttatc aacagccacc aactatctac tcaacgcaat tggggcactt gataatgtga 2341 caatcctcct tgctcctttc cttgtcgcct caaacaacgt cgggaactgc acagaaacat 2401 cgctacttat tcacgcctca cctctctttc ctcctttact cacttattcg cgcctcacct 2461 ctctttcctc ctttactcaa gtcctctgct tgccgggttt attcttccgt ttgtgttgtt 2521 gcggtctgat aggatttgtt tggagacgaa cctttttttt tgattgcttt ataaattaac 2581 gttttgccga aattgttgag acctaagggc gaattcggag cctgcttttt tgtacaaagt 2641 tggcattata aaaaagcatt gctcatcaat ttgttgcaac gaacaggtca ctatcagtca 2701 aaataaaatc attatttggg gcccgagctt aagactggcc gtcgttttac aacgtcgtga 2761 ctgggaaaac atccatgcta gcgttaacgc gagagtaggg aactgccagg catcaaataa 2821 aacgaaaggc tcagtcggaa gactgggcct ttcgttttat ctgttgtttg tcggtgaacg 2881 ctctcctgag taggacaaat ccgccgggag cggatttgaa cgttgtgaag caacggcccg 2941 gagggtggcg ggcaggacgc ccgccataaa ctgccaggca tcaaactaag cagaaggcca 3001 tcctgacgga tggccttttt gcgtttctac aaactcttcc tggctagcgg tacgcgtatt 3061 aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta 3121 atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc 3181 gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa 3241 ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaag //