LOCUS       sequence-84623-e        8304 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
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 Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "PLoS One.";
            date: "2014-02-13"; volume: "9(2):e88218. doi"; pages: " 
            10.1371/journal.pone.0088218. eCollection 2014"
COMMENT     Sequence Label: LIIbeta F 1-2 RNAi
FEATURES             Location/Qualifiers
     source          1..8304
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     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        join(2390..2395,2396..2413,2414..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
//
