LOCUS Exported 8260 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS LIIbeta F 3-4 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 8260) 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 8260) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..8260 /organism="synthetic DNA construct" /mol_type="other DNA" CDS 371..1162 /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" misc_feature 1540..1564 /label=LB T-DNA repeat /note="left border repeat from nopaline C58 T-DNA" promoter 1758..1788 /label=lac UV5 promoter /note="E. coli lac promoter with an ""up"" mutation" CDS 1842..2501 /codon_start=1 /gene="cat" /product="chloramphenicol acetyltransferase" /label=CmR /note="confers resistance to chloramphenicol" /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA" primer_bind complement(1908..1927) /label=CAT-R /note="Chloramphenicol resistance gene, reverse primer" CDS 2843..3148 /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 3027..3046 /label=ccdB-fwd /note="ccdB gene, forward primer" misc_feature 3234..3258 /label=RB T-DNA repeat /note="right border repeat from nopaline C58 T-DNA" CDS 4559..5188 /codon_start=1 /product="stability protein from the Pseudomonas plasmid pVS1 (Heeb et al., 2000)" /label=pVS1 StaA /translation="MKVIAVLNQKGGSGKTTIATHLARALQLAGADVLLVDSDPQGSAR DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI" CDS 5617..6690 /codon_start=1 /product="replication protein from the Pseudomonas plasmid pVS1 (Heeb et al., 2000)" /label=pVS1 RepA /translation="MSGRKPSGPVQIGAALGDDLVEKLKAAQAAQRQRIEAEARPGESW QAAADRIRKESRQPPAAGAPSIRKPPKGDEQPDFFVPMLYDVGTRDSRSIMDVAVFRLS KRDRRAGEVIRYELPDGHVEVSAGPAGMASVWDYDLVLMAVSHLTESMNRYREGKGDKP GRVFRPHVADVLKFCRRADGGKQKDDLVETCIRLNTTHVAMQRTKKAKNGRLVTVSEGE ALISRYKIVKSETGRPEYIEIELADWMYREITEGKNPDVLTVHPDYFLIDPGIGRFLYR LARRAAGKAEARWLFKTIYERSGSAGEFKKFCFTVRKLIGSNDLPEYDLKEEAGQAGPI LVMRYRNLIEGEASAGS" rep_origin 6756..6950 /label=pVS1 oriV /note="origin of replication for the Pseudomonas plasmid pVS1 (Heeb et al., 2000)" primer_bind complement(7186..7208) /label=pGEX 3' /note="pGEX vectors, reverse primer" misc_feature 7294..7434 /label=bom /note="basis of mobility region from pBR322" primer_bind 7349..7368 /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind complement(7449..7466) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(7620..8208) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(7700..7719) /label=pBR322ori-F /note="pBR322 origin, forward primer" ORIGIN 1 actaagctgc cgggtttgaa acacggatga tctcgcggag ggtagcatgt tgattgtaac 61 gatgacagag cgttgctgcc tgtgatcaat tcgggcacga acccagtgga cataagcctg 121 ttcggttcgt aagctgtaat gcaagtagcg tatgcgctca cgcaactggt ccagaacctt 181 gaccgaacgc agcggtggta acggcgcagt ggcggttttc atggcttgtt atgactgttt 241 ttttggggta cagtctatgc ctcgggcatc caagcagcaa gcgcgttacg ccgtgggtcg 301 atgtttgatg ttatggagca gcaacgatgt tacgcagcag ggcagtcgcc ctaaaacaaa 361 gttaaacatc atgggggaag cggtgatcgc cgaagtatcg actcaactat cagaggtagt 421 tggcgtcatc gagcgccatc tcgaaccgac gttgctggcc gtacatttgt acggctccgc 481 agtggatggc ggcctgaagc cacacagtga tattgatttg ctggttacgg tgaccgtaag 541 gcttgatgaa acaacgcggc gagctttgat caacgacctt ttggaaactt cggcttcccc 601 tggagagagc gagattctcc gcgctgtaga agtcaccatt gttgtgcacg acgacatcat 661 tccgtggcgt tatccagcta agcgcgaact gcaatttgga gaatggcagc gcaatgacat 721 tcttgcaggt atcttcgagc cagccacgat cgacattgat ctggctatct tgctgacaaa 781 agcaagagaa catagcgttg ccttggtagg tccagcggcg gaggaactct ttgatccggt 841 tcctgaacag gatctatttg aggcgctaaa tgaaacctta acgctatgga actcgccgcc 901 cgactgggct ggcgatgagc gaaatgtagt gcttacgttg tcccgcattt ggtacagcgc 961 agtaaccggc aaaatcgcgc cgaaggatgt cgctgccgac tgggcaatgg agcgcctgcc 1021 ggcccagtat cagcccgtca tacttgaagc tagacaggct tatcttggac aagaagaaga 1081 tcgcttggcc tcgcgcgcag atcagttgga agaatttgtc cactacgtga aaggcgagat 1141 caccaaggta gtcggcaaat aatgtctagc tagaaattcg ttcaagccga cgccgcttcg 1201 cggcgcggct taactcaagc gttagatgca ctaagcacat aattgctcac agccaaacta 1261 tcaggtcaag tctgctttta ttatttttaa gcgtgcataa taagccctac acaaattggg 1321 agatatatca tgcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga 1381 ccccgtagaa aagatcaaag gatcttctgc cggtaacatg agcaaagtct gccgccttac 1441 aacggctctc ccgctgacgc cgtcccggac tgatgggctg cctgtatcga gtggtgattt 1501 tgtgccgagc tgccggtcgg ggagctgttg gctggctggt ggcaggatat attgtggtgt 1561 aaacaaattg acgcttagac ggattacgga agacaaattg gcggtgagac caaagctgaa 1621 cgagaaacgt aaaatgatat aaatatcaat atattaaatt agattttgca taaaaaacag 1681 actacataat actgtaaaac acaacatatc cagtcactat ggcggcccct cgaggggccg 1741 cattaggcac cccaggcttt acactttatg cttccggctc gtataatgtg tggattttga 1801 gttaggatcc gtcgagattt tcaggagcta aggaagctaa aatggagaaa aaaatcactg 1861 gatataccac cgttgatata tcccaatggc atcgtaaaga acattttgag gcatttcagt 1921 cagttgctca atgtacctat aaccagaccg ttcagctgga tattacggcc tttttaaaga 1981 ccgtaaagaa aaataagcac aagttttatc cggcctttat tcacattctt gcccgcctga 2041 tgaatgctca tccggaattc cgtatggcaa tgaaagacgg tgagctggtg atatgggata 2101 gtgttcaccc ttgttacacc gttttccatg agcaaactga aacgttttca tcgctctgga 2161 gtgaatacca cgacgatttc cggcagtttc tacacatata ttcgcaagat gtggcgtgtt 2221 acggtgaaaa cctggcctat ttccctaaag ggtttattga gaatatgttt ttcgtgtcag 2281 ccaatccctg ggtgagtttc accagttttg atttaaacgt ggccaatatg gacaacttct 2341 tcgcccccgt tttcaccatg ggcaaatatt atacgcaagg cgacaaggtg ctgatgccgc 2401 tggcgattca ggttcatcat gccgtttgtg atggcttcca tgtcggcaga atgcttaatg 2461 aattacaaca gtactgcgat gagtggcagg gcggggcgta atctagagga tccggcttac 2521 taaaagccag ataacagtat gcgtatttgc gcgctgattt ttgcggtata agaatatata 2581 ctgatatgta tacccgaagt atgtcaaaaa gaggtatgct atgaagcagc gtattacagt 2641 gacagttgac agcgacagct atcagttgct caaggcatat atgatgtcaa tatctccggt 2701 ctggtaagca caaccatgca gaatgaagcc cgtcgtctgc gtgccgaacg ctggaaagcg 2761 gaaaatcagg aagggatggc tgaggtcgcc cggtttattg aaatgaacgg ctcttttgct 2821 gacgagaaca ggggctggtg aaatgcagtt taaggtttac acctataaaa gagagagccg 2881 ttatcgtctg tttgtggatg tacagagtga tattattgac acgcccgggc gacggatggt 2941 gatccccctg gccagtgcac gtctgctgtc agataaagtc ccccgtgaac tttacccggt 3001 ggtgcatatc ggggatgaaa gctggcgcat gatgaccacc gatatggcca gtgtgccggt 3061 ttccgttatc ggggaagaag tggctgatct cagccaccgc gaaaatgaca tcaaaaacgc 3121 cattaacctg atgttctggg gaatataaat gtcaggctcc cttatacaca gccagtctgc 3181 aggtcgacgg tctcatgtct gtcttgtctt ctctggttta aactatcagt gtttgacagg 3241 atatattggc gggtaaacct aagagaaaag agcgtttatt agaataatcg gatatttaaa 3301 agggcgtgaa aaggtttatc cgttcgtcca tttgtatgtg catgccaacc acagggttcc 3361 cctcgggatc aaagtacttt gatccaaccc ctccgctgct atagtgcagt cggcttctga 3421 cgttcagtgc agccttcttc tgaaaacgac atgtcgcaca agtcctaagt tacgcgacag 3481 gctgccgccc tgcccttttc ctggcgtttt cttgtcgcgt gttttagtcg cataaagtag 3541 aatacttgcg actagaaccg gagacattac gccatgaaca agagcgccgc cgctggcctg 3601 ctgggctatg cccgcgtcag caccgacgac caggacttga ccaaccaacg ggccgaactg 3661 cacgcggccg gctgcaccaa gctgttttcc gagaagatca ccggcaccag gcgcgaccgc 3721 ccggagctgg ccaggatgct tgaccaccta cgccctggcg acgttgtgac agtgaccagg 3781 ctagaccgcc tggcccgcag cacccgcgac ctactggaca ttgccgagcg catccaggag 3841 gccggcgcgg gcctgcgtag cctggcagag ccgtgggccg acaccaccac gccggccggc 3901 cgcatggtgt tgaccgtgtt cgccggcatt gccgagttcg agcgttccct aatcatcgac 3961 cgcacccgga gcgggcgcga ggccgccaag gcccgaggcg tgaagtttgg cccccgccct 4021 accctcaccc cggcacagat cgcgcacgcc cgcgagctga tcgaccagga aggccgcacc 4081 gtgaaagagg cggctgcact gcttggcgtg catcgctcga ccctgtaccg cgcacttgag 4141 cgcagcgagg aagtgacgcc caccgaggcc aggcggcgcg gtgccttccg tgaggacgca 4201 ttgaccgagg ccgacgccct ggcggccgcc gagaatgaac gccaagagga acaagcatga 4261 aaccgcacca ggacggccag gacgaaccgt ttttcattac cgaagagatc gaggcggaga 4321 tgatcgcggc cgggtacgtg ttcgagccgc ccgcgcacat ctcaaccgtg cggctgcatg 4381 aaatcctggc cggtttgtct gatgccaagc tggcggcctg gccggccagc ttggccgctg 4441 aagaaaccga gcgccgccgt ctaaaaaggt gatgtgtatt tgagtaaaac agcttgcgtc 4501 atgcggtcgc tgcgtatatg atgcgatgag taaataaaca aatacgcaag gggaacgcat 4561 gaaggttatc gctgtactta accagaaagg cgggtcaggc aagacgacca tcgcaaccca 4621 tctagcccgc gccctgcaac tcgccggggc cgatgttctg ttagtcgatt ccgatcccca 4681 gggcagtgcc cgcgattggg cggccgtgcg ggaagatcaa ccgctaaccg ttgtcggcat 4741 cgaccgcccg acgattgacc gcgacgtgaa ggccatcggc cggcgcgact tcgtagtgat 4801 cgacggagcg ccccaggcgg cggacttggc tgtgtccgcg atcaaggcag ccgacttcgt 4861 gctgattccg gtgcagccaa gcccttacga catatgggcc accgccgacc tggtggagct 4921 ggttaagcag cgcattgagg tcacggatgg aaggctacaa gcggcctttg tcgtgtcgcg 4981 ggcgatcaaa ggcacgcgca tcggcggtga ggttgccgag gcgctggccg ggtacgagct 5041 gcccattctt gagtcccgta tcacgcagcg cgtgagctac ccaggcactg ccgccgccgg 5101 cacaaccgtt cttgaatcag aacccgaggg cgacgctgcc cgcgaggtcc aggcgctggc 5161 cgctgaaatt aaatcaaaac tcatttgagt taatgaggta aagagaaaat gagcaaaagc 5221 acaaacacgc taagtgccgg ccgtccgagc gcacgcagca gcaaggctgc aacgttggcc 5281 agcctggcag acacgccagc catgaagcgg gtcaactttc agttgccggc ggaggatcac 5341 accaagctga agatgtacgc ggtacgccaa ggcaagacca ttaccgagct gctatctgaa 5401 tacatcgcgc agctaccaga gtaaatgagc aaatgaataa atgagtagat gaattttagc 5461 ggctaaagga ggcggcatgg aaaatcaaga acaaccaggc accgacgccg tggaatgccc 5521 catgtgtgga ggaacgggcg gttggccagg cgtaagcggc tgggttgcct gccggccctg 5581 caatggcact ggaaccccca agcccgagga atcggcgtga gcggtcgcaa accatccggc 5641 ccggtacaaa tcggcgcggc gctgggtgat gacctggtgg agaagttgaa ggccgcgcag 5701 gccgcccagc ggcaacgcat cgaggcagaa gcacgccccg gtgaatcgtg gcaagcggcc 5761 gctgatcgaa tccgcaaaga atcccggcaa ccgccggcag ccggtgcgcc gtcgattagg 5821 aagccgccca agggcgacga gcaaccagat tttttcgttc cgatgctcta tgacgtgggc 5881 acccgcgata gtcgcagcat catggacgtg gccgttttcc gtctgtcgaa gcgtgaccga 5941 cgagctggcg aggtgatccg ctacgagctt ccagacgggc acgtagaggt ttccgcaggg 6001 ccggccggca tggccagtgt gtgggattac gacctggtac tgatggcggt ttcccatcta 6061 accgaatcca tgaaccgata ccgggaaggg aagggagaca agcccggccg cgtgttccgt 6121 ccacacgttg cggacgtact caagttctgc cggcgagccg atggcggaaa gcagaaagac 6181 gacctggtag aaacctgcat tcggttaaac accacgcacg ttgccatgca gcgtacgaag 6241 aaggccaaga acggccgcct ggtgacggta tccgagggtg aagccttgat tagccgctac 6301 aagatcgtaa agagcgaaac cgggcggccg gagtacatcg agatcgagct agctgattgg 6361 atgtaccgcg agatcacaga aggcaagaac ccggacgtgc tgacggttca ccccgattac 6421 tttttgatcg atcccggcat cggccgtttt ctctaccgcc tggcacgccg cgccgcaggc 6481 aaggcagaag ccagatggtt gttcaagacg atctacgaac gcagtggcag cgccggagag 6541 ttcaagaagt tctgtttcac cgtgcgcaag ctgatcgggt caaatgacct gccggagtac 6601 gatttgaagg aggaggcggg gcaggctggc ccgatcctag tcatgcgcta ccgcaacctg 6661 atcgagggcg aagcatccgc cggttcctaa tgtacggagc agatgctagg gcaaattgcc 6721 ctagcagggg aaaaaggtcg aaaaggactc tttcctgtgg atagcacgta cattgggaac 6781 ccaaagccgt acattgggaa ccggaacccg tacattggga acccaaagcc gtacattggg 6841 aaccggtcac acatgtaagt gactgatata aaagagaaaa aaggcgattt ttccgcctaa 6901 aactctttaa aacttattaa aactcttaaa acccgcctgg cctgtgcata actgtctggc 6961 cagcgcacag ccgaagagct gcaaaaagcg cctacccttc ggtcgctgcg ctccctacgc 7021 cccgccgctt cgcgtcggcc tatcgcggcc gctggccgct caaaaatggc tggcctacgg 7081 ccaggcaatc taccagggcg cggacaagcc gcgccgtcgc cactcgaccg ccggcgccca 7141 catcaaggca ccctgcctcg cgcgtttcgg tgatgacggt gaaaacctct gacacatgca 7201 gctcccggag acagtcacag cttgtctgta agcggatgcc gggagcagac aagcccgtca 7261 gggcgcgtca gcgggtgttg gcgggtgtcg gggcgcagcc atgacccagt cacgtagcga 7321 tagcggagtg tatactggct taactatgcg gcatcagagc agattgtact gagagtgcac 7381 catatgcggt gtgaaatacc gcacagatgc gtaaggagaa aataccgcat caggcgctct 7441 tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca 7501 gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac 7561 atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt 7621 ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg 7681 cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc 7741 tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc 7801 gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc 7861 aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac 7921 tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt 7981 aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct 8041 aactacggct acactagaag gacagtattt ggtatctgcg ctctgctgaa gccagttacc 8101 ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt 8161 ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg 8221 atcttttcta cggggtctga cgctcatcgg aagaacggca //