LOCUS       sequence-84617-e        6350 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 6350)
  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 6350)
  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-K
FEATURES             Location/Qualifiers
     source          1..6350
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    18..158
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     73..92
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(173..190)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(344..932)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(424..443)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     complement(1002..1021)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1003..1021)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1026..1042)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1026..1042)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1144..1163)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             1155..1964
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP
                     DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA
                     FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
                     FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
                     RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     complement(1222..1241)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     misc_feature    2220..2244
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     misc_feature    2308..2332
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     CDS             3633..4262
                     /codon_start=1
                     /product="stability protein from the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
                     /label=pVS1 StaA
                     /translation="MKVIAVLNQKGGSGKTTIATHLARALQLAGADVLLVDSDPQGSAR
                     DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP
                     VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI
                     LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI"
     CDS             4691..5764
                     /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      5830..6024
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
     primer_bind     complement(6260..6282)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
ORIGIN
        1 gttggcgggt gtcggggcgc agccatgacc cagtcacgta gcgatagcgg agtgtatact
       61 ggcttaacta tgcggcatca gagcagattg tactgagagt gcaccatatg cggtgtgaaa
      121 taccgcacag atgcgtaagg agaaaatacc gcatcaggcg ctcttccgct tcctcgctca
      181 ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg
      241 taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc
      301 agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc
      361 cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac
      421 tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc
      481 tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata
      541 gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc
      601 acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca
      661 acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag
      721 cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta
      781 gaaggacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg
      841 gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc
      901 agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt
      961 ctgacgctca taaaatcatt atttgccatc cagctgatat cccctatagt gagtcgtatt
     1021 acatggtcat agctgtttcc tggcagctct ggcccgtgtc tcaaaatctc tgatgttaca
     1081 ttgcacaaga taaaaatata tcatcatgaa caataaaact gtctgcttac ataaacagta
     1141 atacaagggg tgttatgagc catattcaac gggaaacgtc gaggccgcga ttaaattcca
     1201 acatggatgc tgatttatat gggtataaat gggctcgcga taatgtcggg caatcaggtg
     1261 cgacaatcta tcgcttgtat gggaagcccg atgcgccaga gttgtttctg aaacatggca
     1321 aaggtagcgt tgccaatgat gttacagatg agatggtcag actaaactgg ctgacggaat
     1381 ttatgcctct tccgaccatc aagcatttta tccgtactcc tgatgatgca tggttactca
     1441 ccactgcgat ccccggaaaa acagcattcc aggtattaga agaatatcct gattcaggtg
     1501 aaaatattgt tgatgcgctg gcagtgttcc tgcgccggtt gcattcgatt cctgtttgta
     1561 attgtccttt taacagcgat cgcgtatttc gtcttgctca ggcgcaatca cgaatgaata
     1621 acggtttggt tgatgcgagt gattttgatg acgagcgtaa tggctggcct gttgaacaag
     1681 tctggaaaga aatgcataaa cttttgccat tctcaccgga ttcagtcgtc actcatggtg
     1741 atttctcact tgataacctt atttttgacg aggggaaatt aataggttgt attgatgttg
     1801 gacgagtcgg aatcgcagac cgataccagg atcttgccat cctatggaac tgcctcggtg
     1861 agttttctcc ttcattacag aaacggcttt ttcaaaaata tggtattgat aatcctgata
     1921 tgaataaatt gcagtttcat ttgatgctcg atgagttttt ctaatcagaa ttggttaatt
     1981 ggttgtaaca ctggcagagc attacgctga cttgacggga cggcgcaagc tcatgaccaa
     2041 aatcccttaa cgtgagttac gcgtcgttcc actgagcgtc agacccctgc cggtaacatg
     2101 agcaaagtct gccgccttac aacggctctc ccgctgacgc cgtcccggac tgatgggctg
     2161 cctgtatcga gtggtgattt tgtgccgagc tgccggtcgg ggagctgttg gctggctggt
     2221 ggcaggatat attgtggtgt aaacaaattg acgcttagac ggattacgtg agacgatacg
     2281 tctcatctgg tttaaactat cagtgtttga caggatatat tggcgggtaa acctaagaga
     2341 aaagagcgtt tattagaata atcggatatt taaaagggcg tgaaaaggtt tatccgttcg
     2401 tccatttgta tgtgcatgcc aaccacaggg ttcccctcgg gatcaaagta ctttgatcca
     2461 acccctccgc tgctatagtg cagtcggctt ctgacgttca gtgcagcctt cttctgaaaa
     2521 cgacatgtcg cacaagtcct aagttacgcg acaggctgcc gccctgccct tttcctggcg
     2581 ttttcttgtc gcgtgtttta gtcgcataaa gtagaatact tgcgactaga accggagaca
     2641 ttacgccatg aacaagagcg ccgccgctgg cctgctgggc tatgcccgcg tcagcaccga
     2701 cgaccaggac ttgaccaacc aacgggccga actgcacgcg gccggctgca ccaagctgtt
     2761 ttccgagaag atcaccggca ccaggcgcga ccgcccggag ctggccagga tgcttgacca
     2821 cctacgccct ggcgacgttg tgacagtgac caggctagac cgcctggccc gcagcacccg
     2881 cgacctactg gacattgccg agcgcatcca ggaggccggc gcgggcctgc gtagcctggc
     2941 agagccgtgg gccgacacca ccacgccggc cggccgcatg gtgttgaccg tgttcgccgg
     3001 cattgccgag ttcgagcgtt ccctaatcat cgaccgcacc cggagcgggc gcgaggccgc
     3061 caaggcccga ggcgtgaagt ttggcccccg ccctaccctc accccggcac agatcgcgca
     3121 cgcccgcgag ctgatcgacc aggaaggccg caccgtgaaa gaggcggctg cactgcttgg
     3181 cgtgcatcgc tcgaccctgt accgcgcact tgagcgcagc gaggaagtga cgcccaccga
     3241 ggccaggcgg cgcggtgcct tccgtgagga cgcattgacc gaggccgacg ccctggcggc
     3301 cgccgagaat gaacgccaag aggaacaagc atgaaaccgc accaggacgg ccaggacgaa
     3361 ccgtttttca ttaccgaaga gatcgaggcg gagatgatcg cggccgggta cgtgttcgag
     3421 ccgcccgcgc acatctcaac cgtgcggctg catgaaatcc tggccggttt gtctgatgcc
     3481 aagctggcgg cctggccggc cagcttggcc gctgaagaaa ccgagcgccg ccgtctaaaa
     3541 aggtgatgtg tatttgagta aaacagcttg cgtcatgcgg tcgctgcgta tatgatgcga
     3601 tgagtaaata aacaaatacg caaggggaac gcatgaaggt tatcgctgta cttaaccaga
     3661 aaggcgggtc aggcaagacg accatcgcaa cccatctagc ccgcgccctg caactcgccg
     3721 gggccgatgt tctgttagtc gattccgatc cccagggcag tgcccgcgat tgggcggccg
     3781 tgcgggaaga tcaaccgcta accgttgtcg gcatcgaccg cccgacgatt gaccgcgacg
     3841 tgaaggccat cggccggcgc gacttcgtag tgatcgacgg agcgccccag gcggcggact
     3901 tggctgtgtc cgcgatcaag gcagccgact tcgtgctgat tccggtgcag ccaagccctt
     3961 acgacatatg ggccaccgcc gacctggtgg agctggttaa gcagcgcatt gaggtcacgg
     4021 atggaaggct acaagcggcc tttgtcgtgt cgcgggcgat caaaggcacg cgcatcggcg
     4081 gtgaggttgc cgaggcgctg gccgggtacg agctgcccat tcttgagtcc cgtatcacgc
     4141 agcgcgtgag ctacccaggc actgccgccg ccggcacaac cgttcttgaa tcagaacccg
     4201 agggcgacgc tgcccgcgag gtccaggcgc tggccgctga aattaaatca aaactcattt
     4261 gagttaatga ggtaaagaga aaatgagcaa aagcacaaac acgctaagtg ccggccgtcc
     4321 gagcgcacgc agcagcaagg ctgcaacgtt ggccagcctg gcagacacgc cagccatgaa
     4381 gcgggtcaac tttcagttgc cggcggagga tcacaccaag ctgaagatgt acgcggtacg
     4441 ccaaggcaag accattaccg agctgctatc tgaatacatc gcgcagctac cagagtaaat
     4501 gagcaaatga ataaatgagt agatgaattt tagcggctaa aggaggcggc atggaaaatc
     4561 aagaacaacc aggcaccgac gccgtggaat gccccatgtg tggaggaacg ggcggttggc
     4621 caggcgtaag cggctgggtt gcctgccggc cctgcaatgg cactggaacc cccaagcccg
     4681 aggaatcggc gtgagcggtc gcaaaccatc cggcccggta caaatcggcg cggcgctggg
     4741 tgatgacctg gtggagaagt tgaaggccgc gcaggccgcc cagcggcaac gcatcgaggc
     4801 agaagcacgc cccggtgaat cgtggcaagc ggccgctgat cgaatccgca aagaatcccg
     4861 gcaaccgccg gcagccggtg cgccgtcgat taggaagccg cccaagggcg acgagcaacc
     4921 agattttttc gttccgatgc tctatgacgt gggcacccgc gatagtcgca gcatcatgga
     4981 cgtggccgtt ttccgtctgt cgaagcgtga ccgacgagct ggcgaggtga tccgctacga
     5041 gcttccagac gggcacgtag aggtttccgc agggccggcc ggcatggcca gtgtgtggga
     5101 ttacgacctg gtactgatgg cggtttccca tctaaccgaa tccatgaacc gataccggga
     5161 agggaaggga gacaagcccg gccgcgtgtt ccgtccacac gttgcggacg tactcaagtt
     5221 ctgccggcga gccgatggcg gaaagcagaa agacgacctg gtagaaacct gcattcggtt
     5281 aaacaccacg cacgttgcca tgcagcgtac gaagaaggcc aagaacggcc gcctggtgac
     5341 ggtatccgag ggtgaagcct tgattagccg ctacaagatc gtaaagagcg aaaccgggcg
     5401 gccggagtac atcgagatcg agctagctga ttggatgtac cgcgagatca cagaaggcaa
     5461 gaacccggac gtgctgacgg ttcaccccga ttactttttg atcgatcccg gcatcggccg
     5521 ttttctctac cgcctggcac gccgcgccgc aggcaaggca gaagccagat ggttgttcaa
     5581 gacgatctac gaacgcagtg gcagcgccgg agagttcaag aagttctgtt tcaccgtgcg
     5641 caagctgatc gggtcaaatg acctgccgga gtacgatttg aaggaggagg cggggcaggc
     5701 tggcccgatc ctagtcatgc gctaccgcaa cctgatcgag ggcgaagcat ccgccggttc
     5761 ctaatgtacg gagcagatgc tagggcaaat tgccctagca ggggaaaaag gtcgaaaagg
     5821 actctttcct gtggatagca cgtacattgg gaacccaaag ccgtacattg ggaaccggaa
     5881 cccgtacatt gggaacccaa agccgtacat tgggaaccgg tcacacatgt aagtgactga
     5941 tataaaagag aaaaaaggcg atttttccgc ctaaaactct ttaaaactta ttaaaactct
     6001 taaaacccgc ctggcctgtg cataactgtc tggccagcgc acagccgaag agctgcaaaa
     6061 agcgcctacc cttcggtcgc tgcgctccct acgccccgcc gcttcgcgtc ggcctatcgc
     6121 ggccgctggc cgctcaaaaa tggctggcct acggccaggc aatctaccag ggcgcggaca
     6181 agccgcgccg tcgccactcg accgccggcg cccacatcaa ggcaccctgc ctcgcgcgtt
     6241 tcggtgatga cggtgaaaac ctctgacaca tgcagctccc ggagacagtc acagcttgtc
     6301 tgtaagcgga tgccgggagc agacaagccc gtcagggcgc gtcagcgggt
//
