LOCUS       sequence-84603-e        6613 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 6613)
  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 6613)
  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 R A-B
FEATURES             Location/Qualifiers
     source          1..6613
                     /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(3229..3234,3235..3252,3253..3259)
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     CDS             3313..3972
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(3379..3398)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     CDS             4314..4619
                     /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     4498..4517
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     misc_feature    4705..4729
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     rep_origin      complement(4852..5440)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4932..4951)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     complement(5578..5597)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(5579..5597)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(5602..5618)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5602..5618)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5720..5739)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             5731..6540
                     /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(5798..5817)
                     /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 cacttaataa cacattgcgg acgtttttaa tgtactgggg tggattacgg
     3061 gtctcagcgg cagattgtct tcaaagctga acgagaaacg taaaatgata taaatatcaa
     3121 tatattaaat tagattttgc ataaaaaaca gactacataa tactgtaaaa cacaacatat
     3181 ccagtcacta tggcggcccc tcgaggggcc gcattaggca ccccaggctt tacactttat
     3241 gcttccggct cgtataatgt gtggattttg agttaggatc cgtcgagatt ttcaggagct
     3301 aaggaagcta aaatggagaa aaaaatcact ggatatacca ccgttgatat atcccaatgg
     3361 catcgtaaag aacattttga ggcatttcag tcagttgctc aatgtaccta taaccagacc
     3421 gttcagctgg atattacggc ctttttaaag accgtaaaga aaaataagca caagttttat
     3481 ccggccttta ttcacattct tgcccgcctg atgaatgctc atccggaatt ccgtatggca
     3541 atgaaagacg gtgagctggt gatatgggat agtgttcacc cttgttacac cgttttccat
     3601 gagcaaactg aaacgttttc atcgctctgg agtgaatacc acgacgattt ccggcagttt
     3661 ctacacatat attcgcaaga tgtggcgtgt tacggtgaaa acctggccta tttccctaaa
     3721 gggtttattg agaatatgtt tttcgtgtca gccaatccct gggtgagttt caccagtttt
     3781 gatttaaacg tggccaatat ggacaacttc ttcgcccccg ttttcaccat gggcaaatat
     3841 tatacgcaag gcgacaaggt gctgatgccg ctggcgattc aggttcatca tgccgtttgt
     3901 gatggcttcc atgtcggcag aatgcttaat gaattacaac agtactgcga tgagtggcag
     3961 ggcggggcgt aatctagagg atccggctta ctaaaagcca gataacagta tgcgtatttg
     4021 cgcgctgatt tttgcggtat aagaatatat actgatatgt atacccgaag tatgtcaaaa
     4081 agaggtatgc tatgaagcag cgtattacag tgacagttga cagcgacagc tatcagttgc
     4141 tcaaggcata tatgatgtca atatctccgg tctggtaagc acaaccatgc agaatgaagc
     4201 ccgtcgtctg cgtgccgaac gctggaaagc ggaaaatcag gaagggatgg ctgaggtcgc
     4261 ccggtttatt gaaatgaacg gctcttttgc tgacgagaac aggggctggt gaaatgcagt
     4321 ttaaggttta cacctataaa agagagagcc gttatcgtct gtttgtggat gtacagagtg
     4381 atattattga cacgcccggg cgacggatgg tgatccccct ggccagtgca cgtctgctgt
     4441 cagataaagt cccccgtgaa ctttacccgg tggtgcatat cggggatgaa agctggcgca
     4501 tgatgaccac cgatatggcc agtgtgccgg tttccgttat cggggaagaa gtggctgatc
     4561 tcagccaccg cgaaaatgac atcaaaaacg ccattaacct gatgttctgg ggaatataaa
     4621 tgtcaggctc ccttatacac agccagtctg caggtcgacg aagacttcgt atctgtgaga
     4681 cctctggttt aaactatcag tgtttgacag gatatattgg cgggtaaacc taagagaaaa
     4741 gagcgtttat tagaataatc ggatatttaa aagggcgtga aaaggtttat ccgttcgtcc
     4801 atttgtatgt gccaaccaca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag
     4861 gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc
     4921 gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt
     4981 tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct
     5041 ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg
     5101 ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct
     5161 tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat
     5221 tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg
     5281 ctacactaga aggacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa
     5341 aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt
     5401 ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc
     5461 tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt
     5521 atcaaaaagg atcttcacct agatcataaa atcattattt gccatccagc tgatatcccc
     5581 tatagtgagt cgtattacat ggtcatagct gtttcctggc agctctggcc cgtgtctcaa
     5641 aatctctgat gttacattgc acaagataaa aatatatcat catgaacaat aaaactgtct
     5701 gcttacataa acagtaatac aaggggtgtt atgagccata ttcaacggga aacgtcgagg
     5761 ccgcgattaa attccaacat ggatgctgat ttatatgggt ataaatgggc tcgcgataat
     5821 gtcgggcaat caggtgcgac aatctatcgc ttgtatggga agcccgatgc gccagagttg
     5881 tttctgaaac atggcaaagg tagcgttgcc aatgatgtta cagatgagat ggtcagacta
     5941 aactggctga cggaatttat gcctcttccg accatcaagc attttatccg tactcctgat
     6001 gatgcatggt tactcaccac tgcgatcccc ggaaaaacag cattccaggt attagaagaa
     6061 tatcctgatt caggtgaaaa tattgttgat gcgctggcag tgttcctgcg ccggttgcat
     6121 tcgattcctg tttgtaattg tccttttaac agcgatcgcg tatttcgtct tgctcaggcg
     6181 caatcacgaa tgaataacgg tttggttgat gcgagtgatt ttgatgacga gcgtaatggc
     6241 tggcctgttg aacaagtctg gaaagaaatg cataaacttt tgccattctc accggattca
     6301 gtcgtcactc atggtgattt ctcacttgat aaccttattt ttgacgaggg gaaattaata
     6361 ggttgtattg atgttggacg agtcggaatc gcagaccgat accaggatct tgccatccta
     6421 tggaactgcc tcggtgagtt ttctccttca ttacagaaac ggctttttca aaaatatggt
     6481 attgataatc ctgatatgaa taaattgcag tttcatttga tgctcgatga gtttttctaa
     6541 tcagaattgg ttaattggtt gtaacactgg cagagcatta cgctgacttg acgggacggc
     6601 gcaagctcat gac
//
