LOCUS       sequence-84614-e        7956 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 7956)
  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 7956)
  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: LIIIbeta R E-F
FEATURES             Location/Qualifiers
     source          1..7956
                     /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"
     promoter        join(2438..2443,2444..2461,2462..2468)
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     CDS             2522..3181
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(2588..2607)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     CDS             3523..3828
                     /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     3707..3726
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     misc_feature    3914..3938
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     CDS             5239..5868
                     /codon_start=1
                     /product="stability protein from the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
                     /label=pVS1 StaA
                     /translation="MKVIAVLNQKGGSGKTTIATHLARALQLAGADVLLVDSDPQGSAR
                     DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP
                     VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI
                     LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI"
     CDS             6297..7370
                     /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      7436..7630
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
     primer_bind     complement(7866..7888)
                     /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 ggattacggg tctcaaatcc
     2281 agattgtctt caaagctgaa cgagaaacgt aaaatgatat aaatatcaat atattaaatt
     2341 agattttgca taaaaaacag actacataat actgtaaaac acaacatatc cagtcactat
     2401 ggcggcccct cgaggggccg cattaggcac cccaggcttt acactttatg cttccggctc
     2461 gtataatgtg tggattttga gttaggatcc gtcgagattt tcaggagcta aggaagctaa
     2521 aatggagaaa aaaatcactg gatataccac cgttgatata tcccaatggc atcgtaaaga
     2581 acattttgag gcatttcagt cagttgctca atgtacctat aaccagaccg ttcagctgga
     2641 tattacggcc tttttaaaga ccgtaaagaa aaataagcac aagttttatc cggcctttat
     2701 tcacattctt gcccgcctga tgaatgctca tccggaattc cgtatggcaa tgaaagacgg
     2761 tgagctggtg atatgggata gtgttcaccc ttgttacacc gttttccatg agcaaactga
     2821 aacgttttca tcgctctgga gtgaatacca cgacgatttc cggcagtttc tacacatata
     2881 ttcgcaagat gtggcgtgtt acggtgaaaa cctggcctat ttccctaaag ggtttattga
     2941 gaatatgttt ttcgtgtcag ccaatccctg ggtgagtttc accagttttg atttaaacgt
     3001 ggccaatatg gacaacttct tcgcccccgt tttcaccatg ggcaaatatt atacgcaagg
     3061 cgacaaggtg ctgatgccgc tggcgattca ggttcatcat gccgtttgtg atggcttcca
     3121 tgtcggcaga atgcttaatg aattacaaca gtactgcgat gagtggcagg gcggggcgta
     3181 atctagagga tccggcttac taaaagccag ataacagtat gcgtatttgc gcgctgattt
     3241 ttgcggtata agaatatata ctgatatgta tacccgaagt atgtcaaaaa gaggtatgct
     3301 atgaagcagc gtattacagt gacagttgac agcgacagct atcagttgct caaggcatat
     3361 atgatgtcaa tatctccggt ctggtaagca caaccatgca gaatgaagcc cgtcgtctgc
     3421 gtgccgaacg ctggaaagcg gaaaatcagg aagggatggc tgaggtcgcc cggtttattg
     3481 aaatgaacgg ctcttttgct gacgagaaca ggggctggtg aaatgcagtt taaggtttac
     3541 acctataaaa gagagagccg ttatcgtctg tttgtggatg tacagagtga tattattgac
     3601 acgcccgggc gacggatggt gatccccctg gccagtgcac gtctgctgtc agataaagtc
     3661 ccccgtgaac tttacccggt ggtgcatatc ggggatgaaa gctggcgcat gatgaccacc
     3721 gatatggcca gtgtgccggt ttccgttatc ggggaagaag tggctgatct cagccaccgc
     3781 gaaaatgaca tcaaaaacgc cattaacctg atgttctggg gaatataaat gtcaggctcc
     3841 cttatacaca gccagtctgc aggtcgacga agacttcgta tgagtgagac ctctggttta
     3901 aactatcagt gtttgacagg atatattggc gggtaaacct aagagaaaag agcgtttatt
     3961 agaataatcg gatatttaaa agggcgtgaa aaggtttatc cgttcgtcca tttgtatgtg
     4021 catgccaacc acagggttcc cctcgggatc aaagtacttt gatccaaccc ctccgctgct
     4081 atagtgcagt cggcttctga cgttcagtgc agccttcttc tgaaaacgac atgtcgcaca
     4141 agtcctaagt tacgcgacag gctgccgccc tgcccttttc ctggcgtttt cttgtcgcgt
     4201 gttttagtcg cataaagtag aatacttgcg actagaaccg gagacattac gccatgaaca
     4261 agagcgccgc cgctggcctg ctgggctatg cccgcgtcag caccgacgac caggacttga
     4321 ccaaccaacg ggccgaactg cacgcggccg gctgcaccaa gctgttttcc gagaagatca
     4381 ccggcaccag gcgcgaccgc ccggagctgg ccaggatgct tgaccaccta cgccctggcg
     4441 acgttgtgac agtgaccagg ctagaccgcc tggcccgcag cacccgcgac ctactggaca
     4501 ttgccgagcg catccaggag gccggcgcgg gcctgcgtag cctggcagag ccgtgggccg
     4561 acaccaccac gccggccggc cgcatggtgt tgaccgtgtt cgccggcatt gccgagttcg
     4621 agcgttccct aatcatcgac cgcacccgga gcgggcgcga ggccgccaag gcccgaggcg
     4681 tgaagtttgg cccccgccct accctcaccc cggcacagat cgcgcacgcc cgcgagctga
     4741 tcgaccagga aggccgcacc gtgaaagagg cggctgcact gcttggcgtg catcgctcga
     4801 ccctgtaccg cgcacttgag cgcagcgagg aagtgacgcc caccgaggcc aggcggcgcg
     4861 gtgccttccg tgaggacgca ttgaccgagg ccgacgccct ggcggccgcc gagaatgaac
     4921 gccaagagga acaagcatga aaccgcacca ggacggccag gacgaaccgt ttttcattac
     4981 cgaagagatc gaggcggaga tgatcgcggc cgggtacgtg ttcgagccgc ccgcgcacat
     5041 ctcaaccgtg cggctgcatg aaatcctggc cggtttgtct gatgccaagc tggcggcctg
     5101 gccggccagc ttggccgctg aagaaaccga gcgccgccgt ctaaaaaggt gatgtgtatt
     5161 tgagtaaaac agcttgcgtc atgcggtcgc tgcgtatatg atgcgatgag taaataaaca
     5221 aatacgcaag gggaacgcat gaaggttatc gctgtactta accagaaagg cgggtcaggc
     5281 aagacgacca tcgcaaccca tctagcccgc gccctgcaac tcgccggggc cgatgttctg
     5341 ttagtcgatt ccgatcccca gggcagtgcc cgcgattggg cggccgtgcg ggaagatcaa
     5401 ccgctaaccg ttgtcggcat cgaccgcccg acgattgacc gcgacgtgaa ggccatcggc
     5461 cggcgcgact tcgtagtgat cgacggagcg ccccaggcgg cggacttggc tgtgtccgcg
     5521 atcaaggcag ccgacttcgt gctgattccg gtgcagccaa gcccttacga catatgggcc
     5581 accgccgacc tggtggagct ggttaagcag cgcattgagg tcacggatgg aaggctacaa
     5641 gcggcctttg tcgtgtcgcg ggcgatcaaa ggcacgcgca tcggcggtga ggttgccgag
     5701 gcgctggccg ggtacgagct gcccattctt gagtcccgta tcacgcagcg cgtgagctac
     5761 ccaggcactg ccgccgccgg cacaaccgtt cttgaatcag aacccgaggg cgacgctgcc
     5821 cgcgaggtcc aggcgctggc cgctgaaatt aaatcaaaac tcatttgagt taatgaggta
     5881 aagagaaaat gagcaaaagc acaaacacgc taagtgccgg ccgtccgagc gcacgcagca
     5941 gcaaggctgc aacgttggcc agcctggcag acacgccagc catgaagcgg gtcaactttc
     6001 agttgccggc ggaggatcac accaagctga agatgtacgc ggtacgccaa ggcaagacca
     6061 ttaccgagct gctatctgaa tacatcgcgc agctaccaga gtaaatgagc aaatgaataa
     6121 atgagtagat gaattttagc ggctaaagga ggcggcatgg aaaatcaaga acaaccaggc
     6181 accgacgccg tggaatgccc catgtgtgga ggaacgggcg gttggccagg cgtaagcggc
     6241 tgggttgcct gccggccctg caatggcact ggaaccccca agcccgagga atcggcgtga
     6301 gcggtcgcaa accatccggc ccggtacaaa tcggcgcggc gctgggtgat gacctggtgg
     6361 agaagttgaa ggccgcgcag gccgcccagc ggcaacgcat cgaggcagaa gcacgccccg
     6421 gtgaatcgtg gcaagcggcc gctgatcgaa tccgcaaaga atcccggcaa ccgccggcag
     6481 ccggtgcgcc gtcgattagg aagccgccca agggcgacga gcaaccagat tttttcgttc
     6541 cgatgctcta tgacgtgggc acccgcgata gtcgcagcat catggacgtg gccgttttcc
     6601 gtctgtcgaa gcgtgaccga cgagctggcg aggtgatccg ctacgagctt ccagacgggc
     6661 acgtagaggt ttccgcaggg ccggccggca tggccagtgt gtgggattac gacctggtac
     6721 tgatggcggt ttcccatcta accgaatcca tgaaccgata ccgggaaggg aagggagaca
     6781 agcccggccg cgtgttccgt ccacacgttg cggacgtact caagttctgc cggcgagccg
     6841 atggcggaaa gcagaaagac gacctggtag aaacctgcat tcggttaaac accacgcacg
     6901 ttgccatgca gcgtacgaag aaggccaaga acggccgcct ggtgacggta tccgagggtg
     6961 aagccttgat tagccgctac aagatcgtaa agagcgaaac cgggcggccg gagtacatcg
     7021 agatcgagct agctgattgg atgtaccgcg agatcacaga aggcaagaac ccggacgtgc
     7081 tgacggttca ccccgattac tttttgatcg atcccggcat cggccgtttt ctctaccgcc
     7141 tggcacgccg cgccgcaggc aaggcagaag ccagatggtt gttcaagacg atctacgaac
     7201 gcagtggcag cgccggagag ttcaagaagt tctgtttcac cgtgcgcaag ctgatcgggt
     7261 caaatgacct gccggagtac gatttgaagg aggaggcggg gcaggctggc ccgatcctag
     7321 tcatgcgcta ccgcaacctg atcgagggcg aagcatccgc cggttcctaa tgtacggagc
     7381 agatgctagg gcaaattgcc ctagcagggg aaaaaggtcg aaaaggactc tttcctgtgg
     7441 atagcacgta cattgggaac ccaaagccgt acattgggaa ccggaacccg tacattggga
     7501 acccaaagcc gtacattggg aaccggtcac acatgtaagt gactgatata aaagagaaaa
     7561 aaggcgattt ttccgcctaa aactctttaa aacttattaa aactcttaaa acccgcctgg
     7621 cctgtgcata actgtctggc cagcgcacag ccgaagagct gcaaaaagcg cctacccttc
     7681 ggtcgctgcg ctccctacgc cccgccgctt cgcgtcggcc tatcgcggcc gctggccgct
     7741 caaaaatggc tggcctacgg ccaggcaatc taccagggcg cggacaagcc gcgccgtcgc
     7801 cactcgaccg ccggcgccca catcaaggca ccctgcctcg cgcgtttcgg tgatgacggt
     7861 gaaaacctct gacacatgca gctcccggag acagtcacag cttgtctgta agcggatgcc
     7921 gggagcagac aagcccgtca gggcgcgtca gcgggt
//
