LOCUS       sequence-84567-e        6917 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 6917)
  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 6917)
  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: LIIalpha R 1-2
FEATURES             Location/Qualifiers
     source          1..6917
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             446..1237
                     /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      1748..1942
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
     CDS             complement(2008..3081)
                     /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"
     CDS             complement(3510..4139)
                     /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    4403..4427
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     promoter        join(4661..4666,4667..4684,4685..4691)
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     CDS             4745..5404
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(4811..4830)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     CDS             5746..6051
                     /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     5930..5949
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     misc_feature    6137..6161
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     rep_origin      complement(6284..6872)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6364..6383)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc
       61 atcggaagaa cggcaactaa gctgccgggt ttgaaacacg gatgatctcg cggagggtag
      121 catgttgatt gtaacgatga cagagcgttg ctgcctgtga tcaattcggg cacgaaccca
      181 gtggacataa gcctgttcgg ttcgtaagct gtaatgcaag tagcgtatgc gctcacgcaa
      241 ctggtccaga accttgaccg aacgcagcgg tggtaacggc gcagtggcgg ttttcatggc
      301 ttgttatgac tgtttttttg gggtacagtc tatgcctcgg gcatccaagc agcaagcgcg
      361 ttacgccgtg ggtcgatgtt tgatgttatg gagcagcaac gatgttacgc agcagggcag
      421 tcgccctaaa acaaagttaa acatcatggg ggaagcggtg atcgccgaag tatcgactca
      481 actatcagag gtagttggcg tcatcgagcg ccatctcgaa ccgacgttgc tggccgtaca
      541 tttgtacggc tccgcagtgg atggcggcct gaagccacac agtgatattg atttgctggt
      601 tacggtgacc gtaaggcttg atgaaacaac gcggcgagct ttgatcaacg accttttgga
      661 aacttcggct tcccctggag agagcgagat tctccgcgct gtagaagtca ccattgttgt
      721 gcacgacgac atcattccgt ggcgttatcc agctaagcgc gaactgcaat ttggagaatg
      781 gcagcgcaat gacattcttg caggtatctt cgagccagcc acgatcgaca ttgatctggc
      841 tatcttgctg acaaaagcaa gagaacatag cgttgccttg gtaggtccag cggcggagga
      901 actctttgat ccggttcctg aacaggatct atttgaggcg ctaaatgaaa ccttaacgct
      961 atggaactcg ccgcccgact gggctggcga tgagcgaaat gtagtgctta cgttgtcccg
     1021 catttggtac agcgcagtaa ccggcaaaat cgcgccgaag gatgtcgctg ccgactgggc
     1081 aatggagcgc ctgccggccc agtatcagcc cgtcatactt gaagctagac aggcttatct
     1141 tggacaagaa gaagatcgct tggcctcgcg cgcagatcag ttggaagaat ttgtccacta
     1201 cgtgaaaggc gagatcacca aggtagtcgg caaataatgt ctagctagaa attcgttcaa
     1261 gccgacgccg cttcgcggcg cggcttaact caagcgttag atgcactaag cacataattg
     1321 ctcacagcca aactatcagg tcaagtctgc ttttattatt tttaagcgtg cataataagc
     1381 cctacacaaa ttgggagata tatcatgcat gaccaaaatc ccttaacgtg agttttcgtt
     1441 ccactgagcg tcagaccccg tagaaaagat caaaggatct tctgccattt ttggggtgag
     1501 gccgttcgcg gccgaggggc gcagcccctg gggggatggg agctcggtgc cttgatgtgg
     1561 gcgccggcgg tcgagtggcg acggcgcggc ttgtccgcgc cctggtagat tgcctggccg
     1621 taggccagcc atttttgagc ggccagcggc cgcgataggc cgacgcgaag cggcggggcg
     1681 tagggagcgc agcgaccgaa gggtaggcgc tttttgcagc tcttcggctg tgcgctggcc
     1741 agacagttat gcacaggcca ggcgggtttt aagagtttta ataagtttta aagagtttta
     1801 ggcggaaaaa tcgccttttt tctcttttat atcagtcact tacatgtgtg accggttccc
     1861 aatgtacggc tttgggttcc caatgtacgg gttccggttc ccaatgtacg gctttgggtt
     1921 cccaatgtac gtgctatcca caggaaagaa gctttttcga cctttttccc ctgctagggc
     1981 aatttgccct agcatctgct ccgtacatta ggaaccggcg gatgcttcgc cctcgatcag
     2041 gttgcggtag cgcatgacta ggatcgggcc agcctgcccc gcctcctcct tcaaatcgta
     2101 ctccggcagg tcatttgacc cgatcagctt gcgcacggtg aaacagaact tcttgaactc
     2161 tccggcgctg ccactgcgtt cgtagatcgt cttgaacaac catctggctt ctgccttgcc
     2221 tgcggcgcgg cgtgccaggc ggtagagaaa acggccgatg ccgggatcga tcaaaaagta
     2281 atcggggtga accgtcagca cgtccgggtt cttgccttct gtgatctcgc ggtacatcca
     2341 atcagctagc tcgatctcga tgtactccgg ccgcccggtt tcgctcttta cgatcttgta
     2401 gcggctaatc aaggcttcac cctcggatac cgtcaccagg cggccgttct tggccttctt
     2461 cgtacgctgc atggcaacgt gcgtggtgtt taaccgaatg caggtttcta ccaggtcgtc
     2521 tttctgcttt ccgccatcgg ctcgccggca gaacttgagt acgtccgcaa cgtgtggacg
     2581 gaacacgcgg ccgggcttgt ctcccttccc ttcccggtat cggttcatgg attcggttag
     2641 atgggaaacc gccatcagta ccaggtcgta atcccacaca ctggccatgc cggccggccc
     2701 tgcggaaacc tctacgtgcc cgtctggaag ctcgtagcgg atcacctcgc cagctcgtcg
     2761 gtcacgcttc gacagacgga aaacggccac gtccatgatg ctgcgactat cgcgggtgcc
     2821 cacgtcatag agcatcggaa cgaaaaaatc tggttgctcg tcgcccttgg gcggcttcct
     2881 aatcgacgac gcaccggctg ccggcggttg ccgggattct ttgcggattc gatcagcggc
     2941 cgcttgccac gattcaccgg ggcgtgcttc tgcctcgatg cgttgccgct gggcggcctg
     3001 cgcggccttc aacttctcca ccaggtcatc acccagcgcc gcgccgattt gtaccgggcc
     3061 ggatggtttg cgaccgctca cgccgattcc tcgggcttgg gggttccagt gccattgcag
     3121 ggccggcaga caacccagcc gcttacgcct ggccaaccgc ccgttcctcc acacatgggg
     3181 cattccacgg cgtcggtgcc tggttgttct tgattttcca tgccgcctcc tttagccgct
     3241 aaaattcatc tactcattta ttcatttgct catttactct ggtagctgcg cgatgtattc
     3301 agatagcagc tcggtaatgg tcttgccttg gcgtaccgcg tacatcttca gcttggtgtg
     3361 atcctccgcc ggcaactgaa agttgacccg cttcatggct ggcgtgtctg ccaggctggc
     3421 caacgttgca gccttgctgc tgcgtgcgct cggacggccg gcacttagcg tgtttgtgct
     3481 tttgctcatt ttctctttac ctcattaact caaatgagtt ttgatttaat ttcagcggcc
     3541 agcgcctgga cctcgcgggc agcgtcgccc tcgggttctg attcaagaac ggttgtgccg
     3601 gcggcggcag tgcctgggta gctcacgcgc tgcgtgatac gggactcaag aatgggcagc
     3661 tcgtacccgg ccagcgcctc ggcaacctca ccgccgatgc gcgtgccttt gatcgcccgc
     3721 gacacgacaa aggccgcttg tagccttcca tccgtgacct caatgcgctg cttaaccagc
     3781 tccaccaggt cggcggtggc ccatatgtcg taagggcttg gctgcaccgg aatcagcacg
     3841 aagtcggctg ccttgatcgc ggacacagcc aagtccgccg cctggggcgc tccgtcgatc
     3901 actacgaagt cgcgccggcc gatggccttc acgtcgcggt caatcgtcgg gcggtcgatg
     3961 ccgacaacgg ttagcggttg atcttcccgc acggccgccc aatcgcgggc actgccctgg
     4021 ggatcggaat cgactaacag aacatcggcc ccggcgagtt gcagggcgcg ggctagatgg
     4081 gttgcgatgg tcgtcttgcc tgacccgcct ttctggttaa gtacagcgat aaccttcatg
     4141 cgttcccctt gcgtatttgt ttatttactc atcgcatcat atacgcagcg accgcatgac
     4201 gcaagctgtt ttactcaaat acacatcacc tttttagacg gcggcgctcg gtttcttcag
     4261 cggccaagct ggccggccag gccgccagct tggcatcaga caaaccggcc aggatttcat
     4321 gcagccgcac ggctgatggg ctgcctgtat cgagtggtga ttttgtgccg agctgccggt
     4381 cggggagctg ttggctggct ggtggcagga tatattgtgg tgtaaacaaa ttgacgctta
     4441 gacacttaat aacacattgc ggacgttttt aatgtactgg ggtggattac ggaagacaat
     4501 acggacatga gaccaaagct gaacgagaaa cgtaaaatga tataaatatc aatatattaa
     4561 attagatttt gcataaaaaa cagactacat aatactgtaa aacacaacat atccagtcac
     4621 tatggcggcc cctcgagggg ccgcattagg caccccaggc tttacacttt atgcttccgg
     4681 ctcgtataat gtgtggattt tgagttagga tccgtcgaga ttttcaggag ctaaggaagc
     4741 taaaatggag aaaaaaatca ctggatatac caccgttgat atatcccaat ggcatcgtaa
     4801 agaacatttt gaggcatttc agtcagttgc tcaatgtacc tataaccaga ccgttcagct
     4861 ggatattacg gcctttttaa agaccgtaaa gaaaaataag cacaagtttt atccggcctt
     4921 tattcacatt cttgcccgcc tgatgaatgc tcatccggaa ttccgtatgg caatgaaaga
     4981 cggtgagctg gtgatatggg atagtgttca cccttgttac accgttttcc atgagcaaac
     5041 tgaaacgttt tcatcgctct ggagtgaata ccacgacgat ttccggcagt ttctacacat
     5101 atattcgcaa gatgtggcgt gttacggtga aaacctggcc tatttcccta aagggtttat
     5161 tgagaatatg tttttcgtgt cagccaatcc ctgggtgagt ttcaccagtt ttgatttaaa
     5221 cgtggccaat atggacaact tcttcgcccc cgttttcacc atgggcaaat attatacgca
     5281 aggcgacaag gtgctgatgc cgctggcgat tcaggttcat catgccgttt gtgatggctt
     5341 ccatgtcggc agaatgctta atgaattaca acagtactgc gatgagtggc agggcggggc
     5401 gtaatctaga ggatccggct tactaaaagc cagataacag tatgcgtatt tgcgcgctga
     5461 tttttgcggt ataagaatat atactgatat gtatacccga agtatgtcaa aaagaggtat
     5521 gctatgaagc agcgtattac agtgacagtt gacagcgaca gctatcagtt gctcaaggca
     5581 tatatgatgt caatatctcc ggtctggtaa gcacaaccat gcagaatgaa gcccgtcgtc
     5641 tgcgtgccga acgctggaaa gcggaaaatc aggaagggat ggctgaggtc gcccggttta
     5701 ttgaaatgaa cggctctttt gctgacgaga acaggggctg gtgaaatgca gtttaaggtt
     5761 tacacctata aaagagagag ccgttatcgt ctgtttgtgg atgtacagag tgatattatt
     5821 gacacgcccg ggcgacggat ggtgatcccc ctggccagtg cacgtctgct gtcagataaa
     5881 gtcccccgtg aactttaccc ggtggtgcat atcggggatg aaagctggcg catgatgacc
     5941 accgatatgg ccagtgtgcc ggtttccgtt atcggggaag aagtggctga tctcagccac
     6001 cgcgaaaatg acatcaaaaa cgccattaac ctgatgttct ggggaatata aatgtcaggc
     6061 tcccttatac acagccagtc tgcaggtcga cggtctcacc gcgggcttgt cttctctggt
     6121 ttaaactatc agtgtttgac aggatatatt ggcgggtaaa cctaagagaa aagagcgttt
     6181 attagaataa tcggatattt aaaagggcgt gaaaaggttt atccgttcgt ccatttgtat
     6241 gtgccaacca caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc
     6301 cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac
     6361 tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc
     6421 tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata
     6481 gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc
     6541 acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca
     6601 acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag
     6661 cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta
     6721 gaaggacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg
     6781 gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc
     6841 agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt
     6901 ctgacgctca gtggaac
//
