LOCUS Exported 8304 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS LIIbeta F 1-2 RNAi SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 8304) AUTHORS Binder A, Lambert J, Morbitzer R, Popp C, Ott T, Lahaye T, Parniske M TITLE A modular plasmid assembly kit for multigene expression, gene silencing and silencing rescue in plants. JOURNAL PLoS One. 2014 Feb 13;9(2):e88218. doi: 10.1371/journal.pone.0088218. eCollection 2014. PUBMED 24551083 REFERENCE 2 (bases 1 to 8304) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..8304 /organism="synthetic DNA construct" /mol_type="other DNA" CDS complement(108..737) /codon_start=1 /product="stability protein from the Pseudomonas plasmid pVS1 (Heeb et al., 2000)" /label=pVS1 StaA /translation="MKVIAVLNQKGGSGKTTIATHLARALQLAGADVLLVDSDPQGSAR DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI" misc_feature 1001..1025 /label=LB T-DNA repeat /note="left border repeat from nopaline C58 T-DNA" promoter 2390..2420 /label=lac UV5 promoter /note="E. coli lac promoter with an ""up"" mutation" CDS 2474..3133 /codon_start=1 /gene="cat" /product="chloramphenicol acetyltransferase" /label=CmR /note="confers resistance to chloramphenicol" /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA" primer_bind complement(2540..2559) /label=CAT-R /note="Chloramphenicol resistance gene, reverse primer" CDS 3475..3780 /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 VPRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI" primer_bind 3659..3678 /label=ccdB-fwd /note="ccdB gene, forward primer" misc_feature 4122..4146 /label=RB T-DNA repeat /note="right border repeat from nopaline C58 T-DNA" rep_origin complement(4269..4857) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(4349..4368) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS 5348..6139 /codon_start=1 /gene="aadA" /product="aminoglycoside adenylyltransferase (Murphy, 1985)" /label=SmR /note="confers resistance to spectinomycin and streptomycin" /translation="MGEAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK" rep_origin 6650..6844 /label=pVS1 oriV /note="origin of replication for the Pseudomonas plasmid pVS1 (Heeb et al., 2000)" CDS complement(6910..7983) /codon_start=1 /product="replication protein from the Pseudomonas plasmid pVS1 (Heeb et al., 2000)" /label=pVS1 RepA /translation="MSGRKPSGPVQIGAALGDDLVEKLKAAQAAQRQRIEAEARPGESW QAAADRIRKESRQPPAAGASSIRKPPKGDEQPDFFVPMLYDVGTRDSRSIMDVAVFRLS KRDRRAGEVIRYELPDGHVEVSAGPAGMASVWDYDLVLMAVSHLTESMNRYREGKGDKP GRVFRPHVADVLKFCRRADGGKQKDDLVETCIRLNTTHVAMQRTKKAKNGRLVTVSEGE ALISRYKIVKSETGRPEYIEIELADWMYREITEGKNPDVLTVHPDYFLIDPGIGRFLYR LARRAAGKAEARWLFKTIYERSGSAGEFKKFCFTVRKLIGSNDLPEYDLKEEAGQAGPI LVMRYRNLIEGEASAGS" ORIGIN 1 cgtgtctgcc aggctggcca acgttgcagc cttgctgctg cgtgcgctcg gacggccggc 61 acttagcgtg tttgtgcttt tgctcatttt ctctttacct cattaactca aatgagtttt 121 gatttaattt cagcggccag cgcctggacc tcgcgggcag cgtcgccctc gggttctgat 181 tcaagaacgg ttgtgccggc ggcggcagtg cctgggtagc tcacgcgctg cgtgatacgg 241 gactcaagaa tgggcagctc gtacccggcc agcgcctcgg caacctcacc gccgatgcgc 301 gtgcctttga tcgcccgcga cacgacaaag gccgcttgta gccttccatc cgtgacctca 361 atgcgctgct taaccagctc caccaggtcg gcggtggccc atatgtcgta agggcttggc 421 tgcaccggaa tcagcacgaa gtcggctgcc ttgatcgcgg acacagccaa gtccgccgcc 481 tggggcgctc cgtcgatcac tacgaagtcg cgccggccga tggccttcac gtcgcggtca 541 atcgtcgggc ggtcgatgcc gacaacggtt agcggttgat cttcccgcac ggccgcccaa 601 tcgcgggcac tgccctgggg atcggaatcg actaacagaa catcggcccc ggcgagttgc 661 agggcgcggg ctagatgggt tgcgatggtc gtcttgcctg acccgccttt ctggttaagt 721 acagcgataa ccttcatgcg ttccccttgc gtatttgttt atttactcat cgcatcatat 781 acgcagcgac cgcatgacgc aagctgtttt actcaaatac acatcacctt tttagacggc 841 ggcgctcggt ttcttcagcg gccaagctgg ccggccaggc cgccagcttg gcatcagaca 901 aaccggccag gatttcatgc agccgcacgg ctgatgggct gcctgtatcg agtggtgatt 961 ttgtgccgag ctgccggtcg gggagctgtt ggctggctgg tggcaggata tattgtggtg 1021 taaacaaatt gacgcttaga caacttaata acacattgcg gacgttttta atgtactggg 1081 gtggattacg gaagacaata cggcggggag agaggatttt gaggaaataa ttaattgaat 1141 tacttgtatt attgataaag taatttagat aagttgttag tacaacttat tgcaactatg 1201 gatagacaaa aatcacttat tttaagggga gaagtatata acaacttatt ataaaatttc 1261 tgagcagccg gcctcctcca atcaatcata gagagtggag cccattctgg aaaaccaagg 1321 aacccccaac ttgcactggt gcggtggccc aattcaaaaa aaacggggcc cagcaaagta 1381 acgccgttgt aacgatttat caatccaaac tcaaaaggcg gcgagatgcg tttaactccg 1441 tgaaattaac aaaccgccaa caacttgcaa tttgcaacta ccgtttccaa gaagaactca 1501 accacacaac gtatcctatc ccaaaccaca cgcaactagt gacgcgtcat aaggacacgt 1561 gtcacaattt gactggttaa taatttcacc gcttttgcta taaattacct ccaatcccct 1621 tagcttcttc acaattcagt tcccaaccct aacaattctt gttcatatcg cttctctcta 1681 ctttcaaggt atgatccaat ttctctcttc tttctctgta atcctttcgt tgagttttgt 1741 ttccgatcaa tcataggtag ttttcttgtt tcgaagcatg agatctagga attttttgtg 1801 attttccaaa attgagatcg gtttgaaatt gaattttaca gcttgaatct cagatcttgt 1861 tttatcaatg ttttcgatgg cttgcgatgt agatctatga taattgtggt tcagttttgt 1921 taggaatcga tttcggttta gcaattgcag attaattagg gtttccaatt gaattcttca 1981 gatccgttat ggaattatgt caaataattt attcaaattg gaaattattg ttagatccac 2041 tcttaatctg tttgatccaa gcttcaatta gggttttcac ttgtttcaat ttcttgttat 2101 ggattctgat ttatctgttg atgttagatc cactcttaat ctgtttgatc caagcgttaa 2161 ttagggtttt cacttgtttc aatttcgtgt gttggattct gatttatctg ttgttgatgt 2221 gattacagtc tgcacctgag accaaagctg aacgagaaac gtaaaatgat ataaatatca 2281 atatattaaa ttagattttg cataaaaaac agactacata atactgtaaa acacaacata 2341 tccagtcact atggcggccc ctcgaggggc cgcattaggc accccaggct ttacacttta 2401 tgcttccggc tcgtataatg tgtggatttt gagttaggat ccgtcgagat tttcaggagc 2461 taaggaagct aaaatggaga aaaaaatcac tggatatacc accgttgata tatcccaatg 2521 gcatcgtaaa gaacattttg aggcatttca gtcagttgct caatgtacct ataaccagac 2581 cgttcagctg gatattacgg cctttttaaa gaccgtaaag aaaaataagc acaagtttta 2641 tccggccttt attcacattc ttgcccgcct gatgaatgct catccggaat tccgtatggc 2701 aatgaaagac ggtgagctgg tgatatggga tagtgttcac ccttgttaca ccgttttcca 2761 tgagcaaact gaaacgtttt catcgctctg gagtgaatac cacgacgatt tccggcagtt 2821 tctacacata tattcgcaag atgtggcgtg ttacggtgaa aacctggcct atttccctaa 2881 agggtttatt gagaatatgt ttttcgtgtc agccaatccc tgggtgagtt tcaccagttt 2941 tgatttaaac gtggccaata tggacaactt cttcgccccc gttttcacca tgggcaaata 3001 ttatacgcaa ggcgacaagg tgctgatgcc gctggcgatt caggttcatc atgccgtttg 3061 tgatggcttc catgtcggca gaatgcttaa tgaattacaa cagtactgcg atgagtggca 3121 gggcggggcg taatctagag gatccggctt actaaaagcc agataacagt atgcgtattt 3181 gcgcgctgat ttttgcggta taagaatata tactgatatg tatacccgaa gtatgtcaaa 3241 aagaggtatg ctatgaagca gcgtattaca gtgacagttg acagcgacag ctatcagttg 3301 ctcaaggcat atatgatgtc aatatctccg gtctggtaag cacaaccatg cagaatgaag 3361 cccgtcgtct gcgtgccgaa cgctggaaag cggaaaatca ggaagggatg gctgaggtcg 3421 cccggtttat tgaaatgaac ggctcttttg ctgacgagaa caggggctgg tgaaatgcag 3481 tttaaggttt acacctataa aagagagagc cgttatcgtc tgtttgtgga tgtacagagt 3541 gatattattg acacgcccgg gcgacggatg gtgatccccc tggccagtgc acgtctgctg 3601 tcagataaag tcccccgtga actttacccg gtggtgcata tcggggatga aagctggcgc 3661 atgatgacca ccgatatggc cagtgtgccg gtttccgtta tcggggaaga agtggctgat 3721 ctcagccacc gcgaaaatga catcaaaaac gccattaacc tgatgttctg gggaatataa 3781 atgtcaggct cccttataca cagccagtct gcaggtcgac ggtctcaggt gaatccggcc 3841 atgctagagt ccgcaaaaat caccagtctc tctctacaaa tctatctctc tctatttttc 3901 tccagaataa tgtgtgagta gttcccagat aagggaatta gggttcttat agggtttcgc 3961 tcatgtgttg agcatataag aaacccttag tatgtatttg tatttgtaaa atacttctat 4021 caataaaatt tctaattcct aaaaccaaaa tccagtgacc ttgagtgacc taagctagcc 4081 tagtgtcggg cttgtcttct ctggtttaaa ctatcagtgt ttgacaggat atattggcgg 4141 gtaaacctaa gagaaaagag cgtttattag aataatcgga tatttaaaag ggcgtgaaaa 4201 ggtttatccg ttcgtccatt tgtatgtgcc aaccacagga accgtaaaaa ggccgcgttg 4261 ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt 4321 cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc 4381 ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct 4441 tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc 4501 gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta 4561 tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca 4621 gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag 4681 tggtggccta actacggcta cactagaagg acagtatttg gtatctgcgc tctgctgaag 4741 ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt 4801 agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa 4861 gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg 4921 attttggtca tgagattatc aaaaaggatc ttcacctaga tcatcggaag aacggcaact 4981 aagctgccgg gtttgaaaca cggatgatct cgcggagggt agcatgttga ttgtaacgat 5041 gacagagcgt tgctgcctgt gatcaattcg ggcacgaacc cagtggacat aagcctgttc 5101 ggttcgtaag ctgtaatgca agtagcgtat gcgctcacgc aactggtcca gaaccttgac 5161 cgaacgcagc ggtggtaacg gcgcagtggc ggttttcatg gcttgttatg actgtttttt 5221 tggggtacag tctatgcctc gggcatccaa gcagcaagcg cgttacgccg tgggtcgatg 5281 tttgatgtta tggagcagca acgatgttac gcagcagggc agtcgcccta aaacaaagtt 5341 aaacatcatg ggggaagcgg tgatcgccga agtatcgact caactatcag aggtagttgg 5401 cgtcatcgag cgccatctcg aaccgacgtt gctggccgta catttgtacg gctccgcagt 5461 ggatggcggc ctgaagccac acagtgatat tgatttgctg gttacggtga ccgtaaggct 5521 tgatgaaaca acgcggcgag ctttgatcaa cgaccttttg gaaacttcgg cttcccctgg 5581 agagagcgag attctccgcg ctgtagaagt caccattgtt gtgcacgacg acatcattcc 5641 gtggcgttat ccagctaagc gcgaactgca atttggagaa tggcagcgca atgacattct 5701 tgcaggtatc ttcgagccag ccacgatcga cattgatctg gctatcttgc tgacaaaagc 5761 aagagaacat agcgttgcct tggtaggtcc agcggcggag gaactctttg atccggttcc 5821 tgaacaggat ctatttgagg cgctaaatga aaccttaacg ctatggaact cgccgcccga 5881 ctgggctggc gatgagcgaa atgtagtgct tacgttgtcc cgcatttggt acagcgcagt 5941 aaccggcaaa atcgcgccga aggatgtcgc tgccgactgg gcaatggagc gcctgccggc 6001 ccagtatcag cccgtcatac ttgaagctag acaggcttat cttggacaag aagaagatcg 6061 cttggcctcg cgcgcagatc agttggaaga atttgtccac tacgtgaaag gcgagatcac 6121 caaggtagtc ggcaaataat gtctagctag aaattcgttc aagccgacgc cgcttcgcgg 6181 cgcggcttaa ctcaagcgtt agatgcacta agcacataat tgctcacagc caaactatca 6241 ggtcaagtct gcttttatta tttttaagcg tgcataataa gccctacaca aattgggaga 6301 tatatcatgc atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc 6361 cgtagaaaag atcaaaggat cttctgccat ttttggggtg aggccgttcg cggccgaggg 6421 gcgcagcccc tggggggatg ggagctcggt gccttgatgt gggcgccggc ggtcgagtgg 6481 cgacggcgcg gcttgtccgc gccctggtag attgcctggc cgtaggccag ccatttttga 6541 gcggccagcg gccgcgatag gccgacgcga agcggcgggg cgtagggagc gcagcgaccg 6601 aagggtaggc gctttttgca gctcttcggc tgtgcgctgg ccagacagtt atgcacaggc 6661 caggcgggtt ttaagagttt taataagttt taaagagttt taggcggaaa aatcgccttt 6721 tttctctttt atatcagtca cttacatgtg tgaccggttc ccaatgtacg gctttgggtt 6781 cccaatgtac gggttccggt tcccaatgta cggctttggg ttcccaatgt acgtgctatc 6841 cacaggaaag aagctttttc gacctttttc ccctgctagg gcaatttgcc ctagcatctg 6901 ctccgtacat taggaaccgg cggatgcttc gccctcgatc aggttgcggt agcgcatgac 6961 taggatcggg ccagcctgcc ccgcctcctc cttcaaatcg tactccggca ggtcatttga 7021 cccgatcagc ttgcgcacgg tgaaacagaa cttcttgaac tctccggcgc tgccactgcg 7081 ttcgtagatc gtcttgaaca accatctggc ttctgccttg cctgcggcgc ggcgtgccag 7141 gcggtagaga aaacggccga tgccgggatc gatcaaaaag taatcggggt gaaccgtcag 7201 cacgtccggg ttcttgcctt ctgtgatctc gcggtacatc caatcagcta gctcgatctc 7261 gatgtactcc ggccgcccgg tttcgctctt tacgatcttg tagcggctaa tcaaggcttc 7321 accctcggat accgtcacca ggcggccgtt cttggccttc ttcgtacgct gcatggcaac 7381 gtgcgtggtg tttaaccgaa tgcaggtttc taccaggtcg tctttctgct ttccgccatc 7441 ggctcgccgg cagaacttga gtacgtccgc aacgtgtgga cggaacacgc ggccgggctt 7501 gtctcccttc ccttcccggt atcggttcat ggattcggtt agatgggaaa ccgccatcag 7561 taccaggtcg taatcccaca cactggccat gccggccggc cctgcggaaa cctctacgtg 7621 cccgtctgga agctcgtagc ggatcacctc gccagctcgt cggtcacgct tcgacagacg 7681 gaaaacggcc acgtccatga tgctgcgact atcgcgggtg cccacgtcat agagcatcgg 7741 aacgaaaaaa tctggttgct cgtcgccctt gggcggcttc ctaatcgacg acgcaccggc 7801 tgccggcggt tgccgggatt ctttgcggat tcgatcagcg gccgcttgcc acgattcacc 7861 ggggcgtgct tctgcctcga tgcgttgccg ctgggcggcc tgcgcggcct tcaacttctc 7921 caccaggtca tcacccagcg ccgcgccgat ttgtaccggg ccggatggtt tgcgaccgct 7981 cacgccgatt cctcgggctt gggggttcca gtgccattgc agggccggca gacaacccag 8041 ccgcttacgc ctggccaacc gcccgttcct ccacacatgg ggcattccac ggcgtcggtg 8101 cctggttgtt cttgattttc catgccgcct cctttagccg ctaaaattca tctactcatt 8161 tattcatttg ctcatttact ctggtagctg cgcgatgtat tcagatagca gctcggtaat 8221 ggtcttgcct tggcgtaccg cgtacatctt cagcttggtg tgatcctccg ccggcaactg 8281 aaagttgacc cgcttcatgg ctgg //