LOCUS       sequence-482846-        7248 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  A suicide vector for replacing 4th to 15th amino acids of tufB.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7248)
  AUTHORS   Liu MS, Huang TK, Wang YC, Wang SC, Wu CH, Kuo CH, Lai EM
  TITLE     Floral stage optimization and immune evasion enhance 
            Agrobacterium-mediated genome editing in Arabidopsis.
  JOURNAL   New Phytol. 2025 Nov 30. doi: 10.1111/nph.70795.
  PUBMED    41321035
REFERENCE   2  (bases 1 to 7248)
  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: "New Phytol.
            2025 Nov 30. doi: 10.1111/nph.70795."
COMMENT     Sequence Label: EML8553_ pEML41003-tufBpst4-15
FEATURES             Location/Qualifiers
     source          1..7248
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      5..99
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     primer_bind     complement(198..217)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             209..1024
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     complement(1204..1221)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1338..1883)
                     /direction=LEFT
                     /label=p15A ori
                     /note="Plasmids containing the medium-copy-number p15A 
                     origin of replication can be propagated in E. coli cells 
                     that contain a second plasmid with the ColE1 origin.
                     "
     primer_bind     complement(1418..1437)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     oriT            2257..2366
                     /note="incP origin of transfer"
     promoter        2746..3191
                     /gene="sacR"
                     /label=sacB promoter
                     /note="sacB promoter and control region"
     CDS             join(3192..3278,3279..4613)
                     /codon_start=1
                     /gene="Bacillus subtilis sacB"
                     /product="secreted levansucrase that renders bacterial 
                     growth sensitive to sucrose"
                     /label=SacB
                     /note="negative selection marker"
                     /translation="MNIKKFAKQATVLTFTTALLAGGATQAFAKETNQKPYKETYGISH
                     ITRHDMLQIPEQQKNEKYQVPEFDSSTIKNISSAKGLDVWDSWPLQNADGTVANYHGYH
                     IVFALAGDPKNADDTSIYMFYQKVGETSIDSWKNAGRVFKDSDKFDANDSILKDQTQEW
                     SGSATFTSDGKIRLFYTDFSGKHYGKQTLTTAQVNVSASDSSLNINGVEDYKSIFDGDG
                     KTYQNVQQFIDEGNYSSGDNHTLRDPHYVEDKGHKYLVFEANTGTEDGYQGEESLFNKA
                     YYGKSTSFFRQESQKLLQSDKKRTAELANGALGMIELNDDYTLKKVMKPLIASNTVTDE
                     IERANVFKMNGKWYLFTDSRGSKMTIDGITSNDIYMLGYVSNSLTGPYKPLNKTGLVLK
                     MDLDPNDVTFTYSHFAVPQAKGNNVVITSYMTNRGFYADKQSTFAPSFLLNIKGKKTSV
                     VKDSILEQGQLTVNK"
     terminator      complement(4723..4809)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
ORIGIN
        1 cttggactcc tgttgataga tccagtaatg acctcagaac tccatctgga tttgttcaga
       61 acgctcggtt gccgccgggc gttttttatt ggtgagaatc caggcaattg tgtctcaaaa
      121 tctctgatgt tacattgcac aagataaaaa tatatcatca tgaacaataa aactgtctgc
      181 ttacataaac agtaatacaa ggggtgttat gagccatatt cagcgtgaaa cgagctgtag
      241 ccgtccgcgt ctgaacagca acatggatgc ggatctgtat ggctataaat gggcgcgtga
      301 taacgtgggt cagagcggcg cgaccattta tcgtctgtat ggcaaaccgg atgcgccgga
      361 actgtttctg aaacatggca aaggcagcgt ggcgaacgat gtgaccgatg aaatggtgcg
      421 tctgaactgg ctgaccgaat ttatgccgct gccgaccatt aaacatttta ttcgcacccc
      481 ggatgatgcg tggctgctga ccaccgcgat tccgggcaaa accgcgtttc aggtgctgga
      541 agaatatccg gatagcggcg aaaacattgt ggatgcgctg gccgtgtttc tgcgtcgtct
      601 gcatagcatt ccggtgtgca actgcccgtt taacagcgat cgtgtgtttc gtctggccca
      661 ggcgcagagc cgtatgaaca acggcctggt ggatgcgagc gattttgatg atgaacgtaa
      721 cggctggccg gtggaacagg tgtggaaaga aatgcataaa ctgctgccgt ttagcccgga
      781 tagcgtggtg acccacggcg attttagcct ggataacctg attttcgatg aaggcaaact
      841 gattggctgc attgatgtgg gccgtgtggg cattgcggat cgttatcagg atctggccat
      901 tctgtggaac tgcctgggcg aatttagccc gagcctgcaa aaacgtctgt ttcagaaata
      961 tggcattgat aatccggata tgaacaaact gcaatttcat ctgatgctgg atgaattttt
     1021 ctaataatta attgaactat gcgtaacggc aaaagcaccg ccggacatca gcgctagcgg
     1081 agtgtatact ggcttactat gttggcactg atgagggtgt cagtgaagtg cttcatgtgg
     1141 caggagaaaa aaggctgcac cggtgcgtca gcagaatatg tgatacagga tatattccgc
     1201 ttcctcgctc actgactcgc tacgctcggt cgttcgactg cggcgagcgg aaatggctta
     1261 cgaacggggc ggagatttcc tggaagatgc caggaagata cttaacaggg aagtgagagg
     1321 gccgcggcaa agccgttttt ccataggctc cgcccccctg acaagcatca cgaaatctga
     1381 cgctcaaatc agtggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct
     1441 ggcggctccc tcgtgcgctc tcctgttcct gcctttcggt ttaccggtgt cattccgctg
     1501 ttatggccgc gtttgtctca ttccacgcct gacactcagt tccgggtagg cagttcgctc
     1561 caagctggac tgtatgcacg aaccccccgt tcagtccgac cgctgcgcct tatccggtaa
     1621 ctatcgtctt gagtccaacc cggaaagaca tgcaaaagca ccactggcag cagccactgg
     1681 taattgattt agaggagtta gtcttgaagt catgcgccgg ttaaggctaa actgaaagga
     1741 caagttttgg tgactgcgct cctccaagcc agttacctcg gttcaaagag ttggtagctc
     1801 agagaacctt cgaaaaaccg ccctgcaagg cggttttttc gttttcagag caagagatta
     1861 cgcgcagacc aaaacgatct caagaagatc atcttattaa tcagataaaa tatttctagc
     1921 tagagtcgat cttcgccagc agggcgagga tcgtggcatc accgaaccgc gccgtgcgcg
     1981 ggtcgtcggt gagccagagt ttcagcaggc cgcccaggtc gccattgatg cgggccagct
     2041 cgcggacgtg ctcatagtcc acgacgcccg tgattttgta gccctggccg acggccagca
     2101 ggtaggccga caggctcatg ccggccgccg ccgccttttc ctcaatcgct cttcgttcgt
     2161 ctggaaggca gtacaccttg ataggtgggc tgcccttcct ggttggcttg gtttcatcag
     2221 ccatccgctt gccctcatct gttacgccgg cggtagccgg ccagcctcgc agagcaggat
     2281 tcccgttgag caccgccagg tgcgaataag ggacagtgaa gaaggaacac ccgctcgcgg
     2341 gtgggcctac ttcacctatc ctgcccggct gacgccgttg gatacaccaa ggaaagtcta
     2401 cacgaaccct ttggcaaaat cctgtatatc gtgcgaaaaa ggatggatat accgaaaaaa
     2461 tcgctataat gaccccgaag cagggttatg cagcggaaaa gcgctgcttc cctgctgttt
     2521 tgtggaatat ctaccgactg gaaacaggca aatgcaggaa attactgaac tgaggggaca
     2581 ggcgagacac gatgccaaag agctacaccg acgagctggc cgagtgggtt gaatcccgcg
     2641 cggccaagaa gcgccggcgt gatgaggctg cggttgcgtt cctggcggtg agggcggatg
     2701 tcgatcgact ctagctagag gatcgatcct ttttaaccca tttaccacat atacctgccg
     2761 ttcactatta tttagtgaaa tgagatatta tgatattttc tgaattgtga ttaaaaaggc
     2821 aactttatgc ccatgcaaca gaaactataa aaaatacaga gaatgaaaag aaacagatag
     2881 attttttagt tctttaggcc cgtagtctgc aaatcctttt atgattttct atcaaacaaa
     2941 agaggaaaat agaccagttg caatccaaac gagagtctaa tagaatgagg tcgaaaagta
     3001 aatcgcgcgg gtttgttact gataaagcag gcaagaccta aaatgtgtaa agggcaaagt
     3061 gtatactttg gcgtcacccc ttacatattt taggtctttt tttattgtgc gtaactaact
     3121 tgccatcttc aaacaggagg gctggaagaa gcagaccgct aacacagtac ataaaaaagg
     3181 agacatgaac gatgaacatc aaaaagtttg caaaacaagc aacagtatta acctttacta
     3241 ccgcactgct ggcaggaggc gcaactcaag cgtttgcgaa agaaacgaac caaaagccat
     3301 ataaggaaac atacggcatt tcccatatta cacgccatga tatgctgcaa atccctgaac
     3361 agcaaaaaaa tgaaaaatat caagttcctg aattcgattc gtccacaatt aaaaatatct
     3421 cttctgcaaa aggcctggac gtttgggaca gctggccatt acaaaacgct gacggcactg
     3481 tcgcaaacta tcacggctac cacatcgtct ttgcattagc cggagatcct aaaaatgcgg
     3541 atgacacatc gatttacatg ttctatcaaa aagtcggcga aacttctatt gacagctgga
     3601 aaaacgctgg ccgcgtcttt aaagacagcg acaaattcga tgcaaatgat tctatcctaa
     3661 aagaccaaac acaagaatgg tcaggttcag ccacatttac atctgacgga aaaatccgtt
     3721 tattctacac tgatttctcc ggtaaacatt acggcaaaca aacactgaca actgcacaag
     3781 ttaacgtatc agcatcagac agctctttga acatcaacgg tgtagaggat tataaatcaa
     3841 tctttgacgg tgacggaaaa acgtatcaaa atgtacagca gttcatcgat gaaggcaact
     3901 acagctcagg cgacaaccat acgctgagag atcctcacta cgtagaagat aaaggccaca
     3961 aatacttagt atttgaagca aacactggaa ctgaagatgg ctaccaaggc gaagaatctt
     4021 tatttaacaa agcatactat ggcaaaagca catcattctt ccgtcaagaa agtcaaaaac
     4081 ttctgcaaag cgataaaaaa cgcacggctg agttagcaaa cggcgctctc ggtatgattg
     4141 agctaaacga tgattacaca ctgaaaaaag tgatgaaacc gctgattgca tctaacacag
     4201 taacagatga aattgaacgc gcgaacgtct ttaaaatgaa cggcaaatgg tacctgttca
     4261 ctgactcccg cggatcaaaa atgacgattg acggcattac gtctaacgat atttacatgc
     4321 ttggttatgt ttctaattct ttaactggcc catacaagcc gctgaacaaa actggccttg
     4381 tgttaaaaat ggatcttgat cctaacgatg taacctttac ttactcacac ttcgctgtac
     4441 ctcaagcgaa aggaaacaat gtcgtgatta caagctatat gacaaacaga ggattctacg
     4501 cagacaaaca atcaacgttt gcgccaagct tcctgctgaa catcaaaggc aagaaaacat
     4561 ctgttgtcaa agacagcatc cttgaacaag gacaattaac agttaacaaa taaaaacgca
     4621 aaagaaaatg ccgatggccg cggcgttgtg acaatttacc gaacaactcc gcggccggga
     4681 agccgatctc ggcttgaacg aattgcagat ctagggcggc ggatttgtcc tactcaggag
     4741 agcgttcacc gacaaacaac agataaaacg aaaggcccag tctttcgact gagcctttcg
     4801 ttttatttga tgcctgccga attcggatcc ggagttagag cttgatttcc acgtcaacgc
     4861 cagcagcgag gtcgagcttc ataagagcat cgacagtctg cggggtgggg tcaacgatat
     4921 cgagaagacg cttgtgcgtc cgcatttcga actgttcacg gcttttcttg tcaacgtgag
     4981 gagagcggtt gacagtaaac ttttcgatgc gagtggggag aggaaccggg ccgcgaacgc
     5041 tggcgccggt gcgctttgcg gtcgacacga tctcacgggt agaggcatcg aggatccggt
     5101 gatcgaacgc cttgaggcgg atacggatat tctggccggt cattcgtctt gtccttgttc
     5161 tttgttattc gtttccgcac gcggaaacac atgaacttct attgatcgtg gcgccgcaga
     5221 ttttcaaaaa tcgagaggaa tgcccctgca ccccaatcat ttcagtagtc tgtattgcta
     5281 aaacaccgcg ctccgcaatt gcttgcgaat ccagcgggtg agtgtgagtg cgcgtttacg
     5341 cgctttcacc gaatttttca agcactaaaa atgcaacgcc gccaggcttt ttattaaaag
     5401 tccggcgacg gtgttcttga caccgccgcc gggctgaatt ttgaaagtcc aaattactcg
     5461 acgatcgaag caacgatgcc ggcgccgacg gtacggccgc cttcgcggat agcgaagcgc
     5521 agcttttctt ccatcgcgat cggaacgatc agctcgactt caacagtgac gttgtcgcca
     5581 ggcataacca tttccgtgcc ttcaggaagc gaaacgatac cggtaacgtc agtcgtacgg
     5641 aagtagaact gcggacggta gttcgtgaag aacggcgtat gacggccgcc ttcttccttc
     5701 gtcaggatgt aggcttctgc catgaacttc ttgtgcggct tgaccgaacc cggcttgcac
     5761 aggatctgac cacgctcgac gccgtcacgg gtaacgccgc gaacgagtgc accgatgttg
     5821 tcgccggcct ggccctggtc gagcagcttg cggaacattt caacgccggt aacagtcgtc
     5881 ttcgaggtcg gacggatgcc gacgatttcg acttcttcac caaccttgac gataccgcgc
     5941 tcaacgcgac ccgtcacaac cgtaccacga cccgagatcg agaacacgtc ttcgatcggc
     6001 atcaggaacg gctggtcgat cggacgctca ggcgtcggga tgtaggcgtc gacagcagcc
     6061 atcagctcgc ggatcgcgtc ttcaccgatc ttcttgtcag aatcttcaag agcagcaagt
     6121 gccgaaccct tgatgatcgg gatatcgtcg cccgggaagt cgtaggacga cagaagttcg
     6181 cgaacttcaa gctcgacgag ctcgagaagc tcggcgtcgt caacctggtc gaccttgttg
     6241 aggaacacga cgatggccgg aacgccaacc tgacgggcaa gcaggatgtg ctcgcgggtc
     6301 tgtggcatcg ggccgtcggc agccgagcaa accaggatcg cgccgtccat ctgggcagca
     6361 ccggtgatca tgttcttcac atagtcggcg tggccggggc agtcaacgtg ggcgtagtga
     6421 cgcgcaggcg tctcatactc aacgtgcgcc gtcgaaatcg tgataccacg ggccttttcc
     6481 tcaggagcgg cgtcgatttg gtcatacgcc ttgaactcgc cgaaatactt cgtgatcgca
     6541 gcagtcaacg acgtcttgcc gtggtcgacg tgaccgatcg tgccgacgtt gcaatgcggg
     6601 agggaacgat caaatttttc ctttgccatt agtggctctc cattcttgtc ccttgacggg
     6661 aataatcttc aaatctctgg tggggcgtac cccgatcact tctgaccgga gtactttgcc
     6721 tggatttcct gcgccacgtt cgacggaacc ggcgaatagt gatcgaacgt catcgagtac
     6781 tgtgcgcggc cctgcgacat ggagcgcagg ttgtcgacgt acttgaacat gttcgcgagc
     6841 ggaacgtgtg cgctgatcac gatggtgatg ccacgcgatt cctgaccctg gatctgaccg
     6901 cgacgcgagt tcagatcgcc gatcacgtca ccgacgtaat cttcaggggt gacgacttcg
     6961 accttcatca tcggctcaag aagctgtgca ccagccttct ttgctgcttc acggaagcag
     7021 gcacgcgatg cgatttcgaa cgcgagaacc gaggagtcga cgtcgtggta tgcgccgtcg
     7081 atcagggttg ccttgacgcc cagcatcggg aagccagcga gcggaccgga agacagaacg
     7141 ctttcgatac ccttctgaac gcccgggatg tattccttcg gaacagcacc accgacgatc
     7201 ttggattcga acttgaagtc ttcgccttca gggttcggtt cgaacgct
//
