LOCUS       sequence-84606-e        6614 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 6614)
  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 6614)
  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: LIIIalpha F E-F
FEATURES             Location/Qualifiers
     source          1..6614
                     /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"
     promoter        join(3230..3235,3236..3253,3254..3260)
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     CDS             3314..3973
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(3380..3399)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     CDS             4315..4620
                     /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     4499..4518
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     misc_feature    4706..4730
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     rep_origin      complement(4853..5441)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4933..4952)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     complement(5579..5598)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(5580..5598)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(5603..5619)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5603..5619)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5721..5740)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             5732..6541
                     /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(5799..5818)
                     /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 ggtctcaaat ctacgttgtc ttcaaagctg aacgagaaac gtaaaatgat ataaatatca
     3121 atatattaaa ttagattttg cataaaaaac agactacata atactgtaaa acacaacata
     3181 tccagtcact atggcggccc ctcgaggggc cgcattaggc accccaggct ttacacttta
     3241 tgcttccggc tcgtataatg tgtggatttt gagttaggat ccgtcgagat tttcaggagc
     3301 taaggaagct aaaatggaga aaaaaatcac tggatatacc accgttgata tatcccaatg
     3361 gcatcgtaaa gaacattttg aggcatttca gtcagttgct caatgtacct ataaccagac
     3421 cgttcagctg gatattacgg cctttttaaa gaccgtaaag aaaaataagc acaagtttta
     3481 tccggccttt attcacattc ttgcccgcct gatgaatgct catccggaat tccgtatggc
     3541 aatgaaagac ggtgagctgg tgatatggga tagtgttcac ccttgttaca ccgttttcca
     3601 tgagcaaact gaaacgtttt catcgctctg gagtgaatac cacgacgatt tccggcagtt
     3661 tctacacata tattcgcaag atgtggcgtg ttacggtgaa aacctggcct atttccctaa
     3721 agggtttatt gagaatatgt ttttcgtgtc agccaatccc tgggtgagtt tcaccagttt
     3781 tgatttaaac gtggccaata tggacaactt cttcgccccc gttttcacca tgggcaaata
     3841 ttatacgcaa ggcgacaagg tgctgatgcc gctggcgatt caggttcatc atgccgtttg
     3901 tgatggcttc catgtcggca gaatgcttaa tgaattacaa cagtactgcg atgagtggca
     3961 gggcggggcg taatctagag gatccggctt actaaaagcc agataacagt atgcgtattt
     4021 gcgcgctgat ttttgcggta taagaatata tactgatatg tatacccgaa gtatgtcaaa
     4081 aagaggtatg ctatgaagca gcgtattaca gtgacagttg acagcgacag ctatcagttg
     4141 ctcaaggcat atatgatgtc aatatctccg gtctggtaag cacaaccatg cagaatgaag
     4201 cccgtcgtct gcgtgccgaa cgctggaaag cggaaaatca ggaagggatg gctgaggtcg
     4261 cccggtttat tgaaatgaac ggctcttttg ctgacgagaa caggggctgg tgaaatgcag
     4321 tttaaggttt acacctataa aagagagagc cgttatcgtc tgtttgtgga tgtacagagt
     4381 gatattattg acacgcccgg gcgacggatg gtgatccccc tggccagtgc acgtctgctg
     4441 tcagataaag tcccccgtga actttacccg gtggtgcata tcggggatga aagctggcgc
     4501 atgatgacca ccgatatggc cagtgtgccg gtttccgtta tcggggaaga agtggctgat
     4561 ctcagccacc gcgaaaatga catcaaaaac gccattaacc tgatgttctg gggaatataa
     4621 atgtcaggct cccttataca cagccagtct gcaggtcgac gaagactttc tgtgagtgag
     4681 acctctggtt taaactatca gtgtttgaca ggatatattg gcgggtaaac ctaagagaaa
     4741 agagcgttta ttagaataat cggatattta aaagggcgtg aaaaggttta tccgttcgtc
     4801 catttgtatg tgccaaccac aggaaccgta aaaaggccgc gttgctggcg tttttccata
     4861 ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc
     4921 cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg
     4981 ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc
     5041 tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg
     5101 gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc
     5161 ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga
     5221 ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg
     5281 gctacactag aaggacagta tttggtatct gcgctctgct gaagccagtt accttcggaa
     5341 aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg
     5401 tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt
     5461 ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat
     5521 tatcaaaaag gatcttcacc tagatcataa aatcattatt tgccatccag ctgatatccc
     5581 ctatagtgag tcgtattaca tggtcatagc tgtttcctgg cagctctggc ccgtgtctca
     5641 aaatctctga tgttacattg cacaagataa aaatatatca tcatgaacaa taaaactgtc
     5701 tgcttacata aacagtaata caaggggtgt tatgagccat attcaacggg aaacgtcgag
     5761 gccgcgatta aattccaaca tggatgctga tttatatggg tataaatggg ctcgcgataa
     5821 tgtcgggcaa tcaggtgcga caatctatcg cttgtatggg aagcccgatg cgccagagtt
     5881 gtttctgaaa catggcaaag gtagcgttgc caatgatgtt acagatgaga tggtcagact
     5941 aaactggctg acggaattta tgcctcttcc gaccatcaag cattttatcc gtactcctga
     6001 tgatgcatgg ttactcacca ctgcgatccc cggaaaaaca gcattccagg tattagaaga
     6061 atatcctgat tcaggtgaaa atattgttga tgcgctggca gtgttcctgc gccggttgca
     6121 ttcgattcct gtttgtaatt gtccttttaa cagcgatcgc gtatttcgtc ttgctcaggc
     6181 gcaatcacga atgaataacg gtttggttga tgcgagtgat tttgatgacg agcgtaatgg
     6241 ctggcctgtt gaacaagtct ggaaagaaat gcataaactt ttgccattct caccggattc
     6301 agtcgtcact catggtgatt tctcacttga taaccttatt tttgacgagg ggaaattaat
     6361 aggttgtatt gatgttggac gagtcggaat cgcagaccga taccaggatc ttgccatcct
     6421 atggaactgc ctcggtgagt tttctccttc attacagaaa cggctttttc aaaaatatgg
     6481 tattgataat cctgatatga ataaattgca gtttcatttg atgctcgatg agtttttcta
     6541 atcagaattg gttaattggt tgtaacactg gcagagcatt acgctgactt gacgggacgg
     6601 cgcaagctca tgac
//
