LOCUS       sequence-84615-e        5008 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 5008)
  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 5008)
  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: Xpre-K
FEATURES             Location/Qualifiers
     source          1..5008
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      316..510
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
     CDS             complement(576..1649)
                     /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(2078..2707)
                     /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    2971..2995
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     misc_feature    3100..3124
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     rep_origin      complement(3247..3835)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3327..3346)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     complement(3973..3992)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3974..3992)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3997..4013)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3997..4013)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4115..4134)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             4126..4935
                     /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(4193..4212)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
ORIGIN
        1 caaaatccct taacgtgagt tacgcgtcgt tccactgagc gtcagacccc tgccattttt
       61 ggggtgaggc cgttcgcggc cgaggggcgc agcccctggg gggatgggag ctcggtgcct
      121 tgatgtgggc gccggcggtc gagtggcgac ggcgcggctt gtccgcgccc tggtagattg
      181 cctggccgta ggccagccat ttttgagcgg ccagcggccg cgataggccg acgcgaagcg
      241 gcggggcgta gggagcgcag cgaccgaagg gtaggcgctt tttgcagctc ttcggctgtg
      301 cgctggccag acagttatgc acaggccagg cgggttttaa gagttttaat aagttttaaa
      361 gagttttagg cggaaaaatc gccttttttc tcttttatat cagtcactta catgtgtgac
      421 cggttcccaa tgtacggctt tgggttccca atgtacgggt tccggttccc aatgtacggc
      481 tttgggttcc caatgtacgt gctatccaca ggaaagaagc tttttcgacc tttttcccct
      541 gctagggcaa tttgccctag catctgctcc gtacattagg aaccggcgga tgcttcgccc
      601 tcgatcaggt tgcggtagcg catgactagg atcgggccag cctgccccgc ctcctccttc
      661 aaatcgtact ccggcaggtc atttgacccg atcagcttgc gcacggtgaa acagaacttc
      721 ttgaactctc cggcgctgcc actgcgttcg tagatcgtct tgaacaacca tctggcttct
      781 gccttgcctg cggcgcggcg tgccaggcgg tagagaaaac ggccgatgcc gggatcgatc
      841 aaaaagtaat cggggtgaac cgtcagcacg tccgggttct tgccttctgt gatctcgcgg
      901 tacatccaat cagctagctc gatctcgatg tactccggcc gcccggtttc gctctttacg
      961 atcttgtagc ggctaatcaa ggcttcaccc tcggataccg tcaccaggcg gccgttcttg
     1021 gccttcttcg tacgctgcat ggcaacgtgc gtggtgttta accgaatgca ggtttctacc
     1081 aggtcgtctt tctgctttcc gccatcggct cgccggcaga acttgagtac gtccgcaacg
     1141 tgtggacgga acacgcggcc gggcttgtct cccttccctt cccggtatcg gttcatggat
     1201 tcggttagat gggaaaccgc catcagtacc aggtcgtaat cccacacact ggccatgccg
     1261 gccggccctg cggaaacctc tacgtgcccg tctggaagct cgtagcggat cacctcgcca
     1321 gctcgtcggt cacgcttcga cagacggaaa acggccacgt ccatgatgct gcgactatcg
     1381 cgggtgccca cgtcatagag catcggaacg aaaaaatctg gttgctcgtc gcccttgggc
     1441 ggcttcctaa tcgacgacgc accggctgcc ggcggttgcc gggattcttt gcggattcga
     1501 tcagcggccg cttgccacga ttcaccgggg cgtgcttctg cctcgatgcg ttgccgctgg
     1561 gcggcctgcg cggccttcaa cttctccacc aggtcatcac ccagcgccgc gccgatttgt
     1621 accgggccgg atggtttgcg accgctcacg ccgattcctc gggcttgggg gttccagtgc
     1681 cattgcaggg ccggcagaca acccagccgc ttacgcctgg ccaaccgccc gttcctccac
     1741 acatggggca ttccacggcg tcggtgcctg gttgttcttg attttccatg ccgcctcctt
     1801 tagccgctaa aattcatcta ctcatttatt catttgctca tttactctgg tagctgcgcg
     1861 atgtattcag atagcagctc ggtaatggtc ttgccttggc gtaccgcgta catcttcagc
     1921 ttggtgtgat cctccgccgg caactgaaag ttgacccgct tcatggctgg cgtgtctgcc
     1981 aggctggcca acgttgcagc cttgctgctg cgtgcgctcg gacggccggc acttagcgtg
     2041 tttgtgcttt tgctcatttt ctctttacct cattaactca aatgagtttt gatttaattt
     2101 cagcggccag cgcctggacc tcgcgggcag cgtcgccctc gggttctgat tcaagaacgg
     2161 ttgtgccggc ggcggcagtg cctgggtagc tcacgcgctg cgtgatacgg gactcaagaa
     2221 tgggcagctc gtacccggcc agcgcctcgg caacctcacc gccgatgcgc gtgcctttga
     2281 tcgcccgcga cacgacaaag gccgcttgta gccttccatc cgtgacctca atgcgctgct
     2341 taaccagctc caccaggtcg gcggtggccc atatgtcgta agggcttggc tgcaccggaa
     2401 tcagcacgaa gtcggctgcc ttgatcgcgg acacagccaa gtccgccgcc tggggcgctc
     2461 cgtcgatcac tacgaagtcg cgccggccga tggccttcac gtcgcggtca atcgtcgggc
     2521 ggtcgatgcc gacaacggtt agcggttgat cttcccgcac ggccgcccaa tcgcgggcac
     2581 tgccctgggg atcggaatcg actaacagaa catcggcccc ggcgagttgc agggcgcggg
     2641 ctagatgggt tgcgatggtc gtcttgcctg acccgccttt ctggttaagt acagcgataa
     2701 ccttcatgcg ttccccttgc gtatttgttt atttactcat cgcatcatat acgcagcgac
     2761 cgcatgacgc aagctgtttt actcaaatac acatcacctt tttagacggc ggcgctcggt
     2821 ttcttcagcg gccaagctgg ccggccaggc cgccagcttg gcatcagaca aaccggccag
     2881 gatttcatgc agccgcacgg ctgatgggct gcctgtatcg agtggtgatt ttgtgccgag
     2941 ctgccggtcg gggagctgtt ggctggctgg tggcaggata tattgtggtg taaacaaatt
     3001 gacgcttaga caacttaata acacattgcg gacgttttta atgtactggg gtggattacg
     3061 tgagacgata cgtctcatct ggtttaaact atcagtgttt gacaggatat attggcgggt
     3121 aaacctaaga gaaaagagcg tttattagaa taatcggata tttaaaaggg cgtgaaaagg
     3181 tttatccgtt cgtccatttg tatgtgccaa ccacaggaac cgtaaaaagg ccgcgttgct
     3241 ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca
     3301 gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct
     3361 cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc
     3421 gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt
     3481 tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc
     3541 cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc
     3601 cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg
     3661 gtggcctaac tacggctaca ctagaaggac agtatttggt atctgcgctc tgctgaagcc
     3721 agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag
     3781 cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga
     3841 tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat
     3901 tttggtcatg agattatcaa aaaggatctt cacctagatc ataaaatcat tatttgccat
     3961 ccagctgata tcccctatag tgagtcgtat tacatggtca tagctgtttc ctggcagctc
     4021 tggcccgtgt ctcaaaatct ctgatgttac attgcacaag ataaaaatat atcatcatga
     4081 acaataaaac tgtctgctta cataaacagt aatacaaggg gtgttatgag ccatattcaa
     4141 cgggaaacgt cgaggccgcg attaaattcc aacatggatg ctgatttata tgggtataaa
     4201 tgggctcgcg ataatgtcgg gcaatcaggt gcgacaatct atcgcttgta tgggaagccc
     4261 gatgcgccag agttgtttct gaaacatggc aaaggtagcg ttgccaatga tgttacagat
     4321 gagatggtca gactaaactg gctgacggaa tttatgcctc ttccgaccat caagcatttt
     4381 atccgtactc ctgatgatgc atggttactc accactgcga tccccggaaa aacagcattc
     4441 caggtattag aagaatatcc tgattcaggt gaaaatattg ttgatgcgct ggcagtgttc
     4501 ctgcgccggt tgcattcgat tcctgtttgt aattgtcctt ttaacagcga tcgcgtattt
     4561 cgtcttgctc aggcgcaatc acgaatgaat aacggtttgg ttgatgcgag tgattttgat
     4621 gacgagcgta atggctggcc tgttgaacaa gtctggaaag aaatgcataa acttttgcca
     4681 ttctcaccgg attcagtcgt cactcatggt gatttctcac ttgataacct tatttttgac
     4741 gaggggaaat taataggttg tattgatgtt ggacgagtcg gaatcgcaga ccgataccag
     4801 gatcttgcca tcctatggaa ctgcctcggt gagttttctc cttcattaca gaaacggctt
     4861 tttcaaaaat atggtattga taatcctgat atgaataaat tgcagtttca tttgatgctc
     4921 gatgagtttt tctaatcaga attggttaat tggttgtaac actggcagag cattacgctg
     4981 acttgacggg acggcgcaag ctcatgac
//
