LOCUS       sequence-84607-e        6612 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 6612)
  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 6612)
  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 E-F
FEATURES             Location/Qualifiers
     source          1..6612
                     /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(3228..3233,3234..3251,3252..3258)
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     CDS             3312..3971
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(3378..3397)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     CDS             4313..4618
                     /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     4497..4516
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     misc_feature    4704..4728
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     rep_origin      complement(4851..5439)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4931..4950)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     complement(5577..5596)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(5578..5596)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(5601..5617)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5601..5617)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5719..5738)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             5730..6539
                     /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(5797..5816)
                     /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 gtctcaaatc cagattgtct tcaaagctga acgaaaacgt aaaatgatat aaatatcaat
     3121 atattaaatt agattttgca taaaaaacag actacataat actgtaaaac acaacatatc
     3181 cagtcactat ggcggcccct cgaggggccg cattaggcac cccaggcttt acactttatg
     3241 cttccggctc gtataatgtg tggattttga gttaggatcc gtcgagattt tcaggagcta
     3301 aggaagctaa aatggagaaa aaaatcactg gatataccac cgttgatata tcccaatggc
     3361 atcgtaaaga acattttgag gcatttcagt cagttgctca atgtacctat aaccagaccg
     3421 ttcagctgga tattacggcc tttttaaaga ccgtaaagaa aaataagcac aagttttatc
     3481 cggcctttat tcacattctt gcccgcctga tgaatgctca tccggaattc cgtatggcaa
     3541 tgaaagacgg tgagctggtg atatgggata gtgttcaccc ttgttacacc gttttccatg
     3601 agcaaactga aacgttttca tcgctctgga gtgaatacca cgacgatttc cggcagtttc
     3661 tacacatata ttcgcaagat gtggcgtgtt acggtgaaaa cctggcctat ttccctaaag
     3721 ggtttattga gaatatgttt ttcgtgtcag ccaatccctg ggtgagtttc accagttttg
     3781 atttaaacgt ggccaatatg gacaacttct tcgcccccgt tttcaccatg ggcaaatatt
     3841 atacgcaagg cgacaaggtg ctgatgccgc tggcgattca ggttcatcat gccgtttgtg
     3901 atggcttcca tgtcggcaga atgcttaatg aattacaaca gtactgcgat gagtggcagg
     3961 gcggggcgta atctagagga tccggcttac taaaagccag ataacagtat gcgtatttgc
     4021 gcgctgattt ttgcggtata agaatatata ctgatatgta tacccgaagt atgtcaaaaa
     4081 gaggtatgct atgaagcagc gtattacagt gacagttgac agcgacagct atcagttgct
     4141 caaggcatat atgatgtcaa tatctccggt ctggtaagca caaccatgca gaatgaagcc
     4201 cgtcgtctgc gtgccgaacg ctggaaagcg gaaaatcagg aagggatggc tgaggtcgcc
     4261 cggtttattg aaatgaacgg ctcttttgct gacgagaaca ggggctggtg aaatgcagtt
     4321 taaggtttac acctataaaa gagagagccg ttatcgtctg tttgtggatg tacagagtga
     4381 tattattgac acgcccgggc gacggatggt gatccccctg gccagtgcac gtctgctgtc
     4441 agataaagtc ccccgtgaac tttacccggt ggtgcatatc ggggatgaaa gctggcgcat
     4501 gatgaccacc gatatggcca gtgtgccggt ttccgttatc ggggaagaag tggctgatct
     4561 cagccaccgc gaaaatgaca tcaaaaacgc cattaacctg atgttctggg gaatataaat
     4621 gtcaggctcc cttatacaca gccagtctgc aggtcgacga agacttcgta tgagtgagac
     4681 ctctggttta aactatcagt gtttgacagg atatattggc gggtaaacct aagagaaaag
     4741 agcgtttatt agaataatcg gatatttaaa agggcgtgaa aaggtttatc cgttcgtcca
     4801 tttgtatgtg ccaaccacag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg
     4861 ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg
     4921 acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt
     4981 ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt
     5041 tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc
     5101 tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt
     5161 gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt
     5221 agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc
     5281 tacactagaa ggacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa
     5341 agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt
     5401 tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct
     5461 acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta
     5521 tcaaaaagga tcttcaccta gatcataaaa tcattatttg ccatccagct gatatcccct
     5581 atagtgagtc gtattacatg gtcatagctg tttcctggca gctctggccc gtgtctcaaa
     5641 atctctgatg ttacattgca caagataaaa atatatcatc atgaacaata aaactgtctg
     5701 cttacataaa cagtaataca aggggtgtta tgagccatat tcaacgggaa acgtcgaggc
     5761 cgcgattaaa ttccaacatg gatgctgatt tatatgggta taaatgggct cgcgataatg
     5821 tcgggcaatc aggtgcgaca atctatcgct tgtatgggaa gcccgatgcg ccagagttgt
     5881 ttctgaaaca tggcaaaggt agcgttgcca atgatgttac agatgagatg gtcagactaa
     5941 actggctgac ggaatttatg cctcttccga ccatcaagca ttttatccgt actcctgatg
     6001 atgcatggtt actcaccact gcgatccccg gaaaaacagc attccaggta ttagaagaat
     6061 atcctgattc aggtgaaaat attgttgatg cgctggcagt gttcctgcgc cggttgcatt
     6121 cgattcctgt ttgtaattgt ccttttaaca gcgatcgcgt atttcgtctt gctcaggcgc
     6181 aatcacgaat gaataacggt ttggttgatg cgagtgattt tgatgacgag cgtaatggct
     6241 ggcctgttga acaagtctgg aaagaaatgc ataaactttt gccattctca ccggattcag
     6301 tcgtcactca tggtgatttc tcacttgata accttatttt tgacgagggg aaattaatag
     6361 gttgtattga tgttggacga gtcggaatcg cagaccgata ccaggatctt gccatcctat
     6421 ggaactgcct cggtgagttt tctccttcat tacagaaacg gctttttcaa aaatatggta
     6481 ttgataatcc tgatatgaat aaattgcagt ttcatttgat gctcgatgag tttttctaat
     6541 cagaattggt taattggttg taacactggc agagcattac gctgacttga cgggacggcg
     6601 caagctcatg ac
//
