LOCUS       sequence-84618-e        6654 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 6654)
  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 6654)
  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: Xpre2-S
FEATURES             Location/Qualifiers
     source          1..6654
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     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"
     misc_feature    1628..1652
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     CDS             2953..3582
                     /codon_start=1
                     /product="stability protein from the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
                     /label=pVS1 StaA
                     /translation="MKVIAVLNQKGGSGKTTIATHLARALQLAGADVLLVDSDPQGSAR
                     DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP
                     VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI
                     LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI"
     CDS             4011..5084
                     /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      5150..5344
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
     primer_bind     complement(5580..5602)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    5688..5828
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     5743..5762
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(5843..5860)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6014..6602)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6094..6113)
                     /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 ggattacgtg agacgatacg tctcatctgg tttaaactat
     1621 cagtgtttga caggatatat tggcgggtaa acctaagaga aaagagcgtt tattagaata
     1681 atcggatatt taaaagggcg tgaaaaggtt tatccgttcg tccatttgta tgtgcatgcc
     1741 aaccacaggg ttcccctcgg gatcaaagta ctttgatcca acccctccgc tgctatagtg
     1801 cagtcggctt ctgacgttca gtgcagcctt cttctgaaaa cgacatgtcg cacaagtcct
     1861 aagttacgcg acaggctgcc gccctgccct tttcctggcg ttttcttgtc gcgtgtttta
     1921 gtcgcataaa gtagaatact tgcgactaga accggagaca ttacgccatg aacaagagcg
     1981 ccgccgctgg cctgctgggc tatgcccgcg tcagcaccga cgaccaggac ttgaccaacc
     2041 aacgggccga actgcacgcg gccggctgca ccaagctgtt ttccgagaag atcaccggca
     2101 ccaggcgcga ccgcccggag ctggccagga tgcttgacca cctacgccct ggcgacgttg
     2161 tgacagtgac caggctagac cgcctggccc gcagcacccg cgacctactg gacattgccg
     2221 agcgcatcca ggaggccggc gcgggcctgc gtagcctggc agagccgtgg gccgacacca
     2281 ccacgccggc cggccgcatg gtgttgaccg tgttcgccgg cattgccgag ttcgagcgtt
     2341 ccctaatcat cgaccgcacc cggagcgggc gcgaggccgc caaggcccga ggcgtgaagt
     2401 ttggcccccg ccctaccctc accccggcac agatcgcgca cgcccgcgag ctgatcgacc
     2461 aggaaggccg caccgtgaaa gaggcggctg cactgcttgg cgtgcatcgc tcgaccctgt
     2521 accgcgcact tgagcgcagc gaggaagtga cgcccaccga ggccaggcgg cgcggtgcct
     2581 tccgtgagga cgcattgacc gaggccgacg ccctggcggc cgccgagaat gaacgccaag
     2641 aggaacaagc atgaaaccgc accaggacgg ccaggacgaa ccgtttttca ttaccgaaga
     2701 gatcgaggcg gagatgatcg cggccgggta cgtgttcgag ccgcccgcgc acatctcaac
     2761 cgtgcggctg catgaaatcc tggccggttt gtctgatgcc aagctggcgg cctggccggc
     2821 cagcttggcc gctgaagaaa ccgagcgccg ccgtctaaaa aggtgatgtg tatttgagta
     2881 aaacagcttg cgtcatgcgg tcgctgcgta tatgatgcga tgagtaaata aacaaatacg
     2941 caaggggaac gcatgaaggt tatcgctgta cttaaccaga aaggcgggtc aggcaagacg
     3001 accatcgcaa cccatctagc ccgcgccctg caactcgccg gggccgatgt tctgttagtc
     3061 gattccgatc cccagggcag tgcccgcgat tgggcggccg tgcgggaaga tcaaccgcta
     3121 accgttgtcg gcatcgaccg cccgacgatt gaccgcgacg tgaaggccat cggccggcgc
     3181 gacttcgtag tgatcgacgg agcgccccag gcggcggact tggctgtgtc cgcgatcaag
     3241 gcagccgact tcgtgctgat tccggtgcag ccaagccctt acgacatatg ggccaccgcc
     3301 gacctggtgg agctggttaa gcagcgcatt gaggtcacgg atggaaggct acaagcggcc
     3361 tttgtcgtgt cgcgggcgat caaaggcacg cgcatcggcg gtgaggttgc cgaggcgctg
     3421 gccgggtacg agctgcccat tcttgagtcc cgtatcacgc agcgcgtgag ctacccaggc
     3481 actgccgccg ccggcacaac cgttcttgaa tcagaacccg agggcgacgc tgcccgcgag
     3541 gtccaggcgc tggccgctga aattaaatca aaactcattt gagttaatga ggtaaagaga
     3601 aaatgagcaa aagcacaaac acgctaagtg ccggccgtcc gagcgcacgc agcagcaagg
     3661 ctgcaacgtt ggccagcctg gcagacacgc cagccatgaa gcgggtcaac tttcagttgc
     3721 cggcggagga tcacaccaag ctgaagatgt acgcggtacg ccaaggcaag accattaccg
     3781 agctgctatc tgaatacatc gcgcagctac cagagtaaat gagcaaatga ataaatgagt
     3841 agatgaattt tagcggctaa aggaggcggc atggaaaatc aagaacaacc aggcaccgac
     3901 gccgtggaat gccccatgtg tggaggaacg ggcggttggc caggcgtaag cggctgggtt
     3961 gcctgccggc cctgcaatgg cactggaacc cccaagcccg aggaatcggc gtgagcggtc
     4021 gcaaaccatc cggcccggta caaatcggcg cggcgctggg tgatgacctg gtggagaagt
     4081 tgaaggccgc gcaggccgcc cagcggcaac gcatcgaggc agaagcacgc cccggtgaat
     4141 cgtggcaagc ggccgctgat cgaatccgca aagaatcccg gcaaccgccg gcagccggtg
     4201 cgccgtcgat taggaagccg cccaagggcg acgagcaacc agattttttc gttccgatgc
     4261 tctatgacgt gggcacccgc gatagtcgca gcatcatgga cgtggccgtt ttccgtctgt
     4321 cgaagcgtga ccgacgagct ggcgaggtga tccgctacga gcttccagac gggcacgtag
     4381 aggtttccgc agggccggcc ggcatggcca gtgtgtggga ttacgacctg gtactgatgg
     4441 cggtttccca tctaaccgaa tccatgaacc gataccggga agggaaggga gacaagcccg
     4501 gccgcgtgtt ccgtccacac gttgcggacg tactcaagtt ctgccggcga gccgatggcg
     4561 gaaagcagaa agacgacctg gtagaaacct gcattcggtt aaacaccacg cacgttgcca
     4621 tgcagcgtac gaagaaggcc aagaacggcc gcctggtgac ggtatccgag ggtgaagcct
     4681 tgattagccg ctacaagatc gtaaagagcg aaaccgggcg gccggagtac atcgagatcg
     4741 agctagctga ttggatgtac cgcgagatca cagaaggcaa gaacccggac gtgctgacgg
     4801 ttcaccccga ttactttttg atcgatcccg gcatcggccg ttttctctac cgcctggcac
     4861 gccgcgccgc aggcaaggca gaagccagat ggttgttcaa gacgatctac gaacgcagtg
     4921 gcagcgccgg agagttcaag aagttctgtt tcaccgtgcg caagctgatc gggtcaaatg
     4981 acctgccgga gtacgatttg aaggaggagg cggggcaggc tggcccgatc ctagtcatgc
     5041 gctaccgcaa cctgatcgag ggcgaagcat ccgccggttc ctaatgtacg gagcagatgc
     5101 tagggcaaat tgccctagca ggggaaaaag gtcgaaaagg actctttcct gtggatagca
     5161 cgtacattgg gaacccaaag ccgtacattg ggaaccggaa cccgtacatt gggaacccaa
     5221 agccgtacat tgggaaccgg tcacacatgt aagtgactga tataaaagag aaaaaaggcg
     5281 atttttccgc ctaaaactct ttaaaactta ttaaaactct taaaacccgc ctggcctgtg
     5341 cataactgtc tggccagcgc acagccgaag agctgcaaaa agcgcctacc cttcggtcgc
     5401 tgcgctccct acgccccgcc gcttcgcgtc ggcctatcgc ggccgctggc cgctcaaaaa
     5461 tggctggcct acggccaggc aatctaccag ggcgcggaca agccgcgccg tcgccactcg
     5521 accgccggcg cccacatcaa ggcaccctgc ctcgcgcgtt tcggtgatga cggtgaaaac
     5581 ctctgacaca tgcagctccc ggagacagtc acagcttgtc tgtaagcgga tgccgggagc
     5641 agacaagccc gtcagggcgc gtcagcgggt gttggcgggt gtcggggcgc agccatgacc
     5701 cagtcacgta gcgatagcgg agtgtatact ggcttaacta tgcggcatca gagcagattg
     5761 tactgagagt gcaccatatg cggtgtgaaa taccgcacag atgcgtaagg agaaaatacc
     5821 gcatcaggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc
     5881 ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata
     5941 acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg
     6001 cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct
     6061 caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa
     6121 gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc
     6181 tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt
     6241 aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg
     6301 ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg
     6361 cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct
     6421 tgaagtggtg gcctaactac ggctacacta gaaggacagt atttggtatc tgcgctctgc
     6481 tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg
     6541 ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc
     6601 aagaagatcc tttgatcttt tctacggggt ctgacgctca tcggaagaac ggca
//
