LOCUS       sequence-182150-        6371 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Transformation, Type: non-T-DNA, Plant Selection: PvUbi2:bar, Viral 
            Replication: none.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6371)
  AUTHORS   Cermak T, Curtin SJ, Gil-Humanes J, Cegan R, Kono TJY, Konecna E, 
            Belanto JJ, Starker CG, Mathre JW, Greenstein RL, Voytas DF
  TITLE     A multi-purpose toolkit to enable advanced genome engineering in 
            plants.
  JOURNAL   Plant Cell. 2017 May 18. pii: tpc.00922.2016. doi: 
            10.1105/tpc.16.00922.
  PUBMED    28522548
REFERENCE   2  (bases 1 to 6371)
  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: "Plant Cell.
            2017 May 18. pii: tpc.00922.2016. doi: 10.1105/tpc.16.00922."
COMMENT     Sequence Label: pTRANS_160
FEATURES             Location/Qualifiers
     source          1..6371
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      84..672
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     573..592
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     826..843
                     /label=L4440
                     /note="L4440 vector, forward primer"
     polyA_signal    876..1050
                     /label=CaMV poly(A) signal
                     /note="cauliflower mosaic virus polyadenylation signal"
     CDS             complement(1057..1608)
                     /codon_start=1
                     /gene="Streptomyces hygroscopicus bar"
                     /product="phosphinothricin acetyltransferase"
                     /label=BlpR
                     /note="confers resistance to bialophos or phosphinothricin"
                     /translation="MSPERRPADIRRATEADMPAVCTIVNHYIETSTVNFRTEPQEPQE
                     WTDDLVRLRERYPWLVAEVDGEVAGIAYAGPWKARNAYDWTAESTVYVSPRHQRTGLGS
                     TLYTHLLKSLEAQGFKSVVAVIGLPNDPSVRMHEALGYAPRGMLRAAGFKHGNWHDVGF
                     WQLDFSLPVPPRPVLPVTEI"
     protein_bind    3499..3623
                     /gene="mutant version of attR"
                     /label=attR1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        join(3648..3653,3654..3671,3672..3678)
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     CDS             3732..4412
                     /codon_start=1
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQAGRNLE
                     DPAY"
     CDS             4732..5037
                     /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
                     VPRELYPVVHIGDESWRMMTTDMASVPASVIGEEVADLSHRENDIKNAINLMFWGI"
     primer_bind     4916..4935
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     protein_bind    complement(5078..5202)
                     /gene="mutant version of attR"
                     /label=attR2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             5571..6362
                     /codon_start=1
                     /gene="aadA"
                     /product="aminoglycoside adenylyltransferase (Murphy, 
                     1985)"
                     /label=SmR
                     /note="confers resistance to spectinomycin and 
                     streptomycin"
                     /translation="MREAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH
                     SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK
                     RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF
                     EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP
                     VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK"
ORIGIN
        1 cacccatgac caaaatccct taacgtgagt tacgcgtcgt tccactgagc gtcagacccc
       61 gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg
      121 caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact
      181 ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt tcttctagtg
      241 tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg
      301 ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac
      361 tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca
      421 cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga
      481 gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc
      541 ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct
      601 gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg
      661 agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct
      721 tttgctcaca tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc
      781 tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc
      841 gaggaagcgg gagagcgccc atatgcaatt cgggggatct ggattttagt actggatttt
      901 ggttttagga attagaaatt ttattgatag aagtatttta caaatacaaa tacatactaa
      961 gggtttctta tatgctcaac acatgagcga aaccctatag gaaccctaat tcccttatct
     1021 gggaactact cacacattat tatggagaaa ctcgagtcaa atctcggtga cgggcaggac
     1081 cggacggggc ggtaccggca ggctgaagtc cagctgccag aaacccacgt catgccagtt
     1141 cccgtgcttg aagccggccg cccgcagcat gccgcggggg gcatatccga gcgcctcgtg
     1201 catgcgcacg ctcgggtcgt tgggcagccc gatgacagcg accacgctct tgaagccctg
     1261 tgcctccagg gacttcagca gatgggtgta gagcgtggag cccagtcccg tccgctggtg
     1321 gcgggggctg acgtacacgg tggactcggc cgtccagtcg taggcgttgc gtgccttcca
     1381 gggacccgcg taggcgatgc cggcgacctc gccgtccacc tcggcgacga gccagggata
     1441 gcgctcccgc agacggacga ggtcgtccgt ccactcctgc ggttcctgcg gctcggtacg
     1501 gaagttgacc gtgcttgtct cgatgtagtg gttgacgatg gtgcagaccg ccggcatgtc
     1561 cgcctcggtg gcacggcgga tgtcggccgg gcgtcgttct gggctcataa gcttctgcaa
     1621 aagagaacca gacaacaggg taagtgccta gcagtaaaca aacagaactc atcacaagca
     1681 aacagcaaca tcatattcat accaacaggt catgtgtgtt catcacatca ttagtactaa
     1741 gcatgccatc atccaagtat atcaaagtaa gggcaaagag catgcatgat catcaggtgc
     1801 acaaaagaat catcaaattg tagcagtaca tatcttcatc tatcatgcat atctatccat
     1861 aacaggacga tgcatgttga ccaggtaaaa gctacaggat cctataggaa cagcagcgta
     1921 tatatcttca ccaatcttag catcttaatc atgtggcaca tgcagtttca atttaagcac
     1981 atgagctagt tgattatgag gtaccagaga atcatcaaac tgatgtagca gcatatatcc
     2041 tcatctatca tgcaatctaa tctaatctaa ctaaacagga aaggtgtgct attcagttaa
     2101 aagctaccgc atcatataca aacggcagca taagaaaaag cataatcatc ttaatcatgc
     2161 aacaaacgca gattcataat aagcccaaga gctagcttgt gatgatctta ttctactctg
     2221 atctacagca atcagataac gacctaacct tgcacatggc aacaaaacaa tcgatcggac
     2281 gaatcagttg tttgttccta gctagcacca tcgaaccaga taatagatgc acgtacagat
     2341 cccgaaaacg aacccaaaaa cagggcagac ctagctgaac ctaggcagcg acccagcaga
     2401 tcgtgagaac gatctcatct acgaacagcc tagaagcaac cccacgattc ccggacaaac
     2461 gacctaaaat ccccacaaat cacatgagca tgacaggata aacagcggaa ccgatcagat
     2521 ctacacgaaa accccacctc ccagccaccc acgatcagga aacacgcgga tctagcatga
     2581 tttcgtcaac gcctcagcct agttcctagc cacagaccaa gcagaaccac caaaccacgc
     2641 cgagcgagga gatggggcaa gaggacgggg gagacgatcg ccgtaccttg aagcggggga
     2701 aggatcgccg agggtcgcga ggagagcaat ttggatttgg aacccggggg ttgtgcgctc
     2761 cgaacgatga gacgatgtga gattgtggga agaggcgcgg agggccctgt atttatgggc
     2821 tgcgacgggg ggaggagagg tggggagggt tggggaagga atcccccacc cgtgccgtga
     2881 cggttccggg ccgtgtgaga ggagcccgct cgtctccgcc acgcaatttc cgcgatcgga
     2941 gcggagcttt cgagaggcgg ctggatggtt ggtggccgtt agatttgtag acgccgttaa
     3001 cgcctcgcct ccaccgggaa gagttttgag cagccgctta tgacaatggc ttaacgacgt
     3061 tagacggagc gttagtggca ggccatgata gaatagacgt attgcaatgg gattataatt
     3121 aaataaataa gaatataata agatatggca agtcggcact catgacatgg tcttcgaaat
     3181 gatagtgctc actttcttag ccgagaaagt tgacgcgact gatttagaag ttaagatatt
     3241 atttctctct tcttttcttt cctcgtcata taaggatgaa ataaacttta gagattgccg
     3301 gtagtgattt tggatttcgg cgatcaggct tggtttgccg gtttcggacg gtgtgcctta
     3361 ggccacccgc agtgtatctt gtaatgttca accgataagc aagggtgggg ctcaagcaag
     3421 tagtaaacaa ctatgtcaaa tgtcaccatg gttatggtct tgtttagttg gcttcggcgc
     3481 gccgatcaaa agcaggtgac aagtttgtac aaaaaagctg aacgagaaac gtaaaatgat
     3541 ataaatatca atatattaaa ttagattttg cataaaaaac agactacata atactgtaaa
     3601 acacaacata tccagtcact atggcggccg cattaggcac cccaggcttt acactttatg
     3661 cttccggctc gtataatgtg tggattttga gttaggatcc gtcgagattt tcaggagcta
     3721 aggaagctaa aatggagaaa aaaatcactg gatataccac cgttgatata tcccaatggc
     3781 atcgtaaaga acattttgag gcatttcagt cagtcgctca atgtacctat aaccagaccg
     3841 ttcagctgga tattacggcc tttttaaaga ccgtaaagaa aaataagcac aagttttatc
     3901 cggcctttat tcacattctt gcccgcctga tgaatgctca tccggaattc cgtatggcaa
     3961 tgaaagacgg tgagctggtg atatgggata gtgttcaccc ttgttacacc gttttccatg
     4021 agcaaactga aacgttttca tcgctctgga gtgaatacca cgacgatttc cggcagtttc
     4081 tacacatata ttcgcaagat gtggcgtgtt acggtgaaaa cctggcctat ttccctaaag
     4141 ggtttattga gaatatgttt ttcgtctcag ccaatccctg ggtgagtttc accagttttg
     4201 atttaaacgt ggccaatatg gacaacttct tcgcccccgt tttcaccatg ggcaaatatt
     4261 atacgcaagg cgacaaggtg ctgatgccgc tggcgattca ggttcatcat gccgtttgtg
     4321 atggcttcca tgtcggcaga atgcttaatg aattacaaca gtactgcgat gagtggcagg
     4381 cggggcgtaa tctagaggat ccggcttact aaaagccaga taacagtatg cgtatttgcg
     4441 cgctgatttt tgcggtataa gaatatatac tgatatgtat accagaagta tgtcaaaaag
     4501 aggtatgcta tgaagcagcg tattacagtg acagttgaca gcgacagcta tcagttgctc
     4561 aaggcatata tgatgtcaat atctccggtc tggtaagcac aaccatgcag aatgaagccc
     4621 gtcgtctgcg tgccgaacgc tggaaagcgg aaaatcagga agggatggct gaggtcgccc
     4681 ggtttattga aatgaacggc tcttttgctg acgagaacag gggctggtga aatgcagttt
     4741 aaggtttaca cctataaaag agagagccgt tatcgtctgt ttgtggatgt acagagtgat
     4801 attattgaca cgcccgggcg acggatggtg atccccctgg ccagtgcacg tctgctgtca
     4861 gataaagtcc cccgtgaact ttacccggtg gtgcatatcg gggatgaaag ctggcgcatg
     4921 atgaccaccg atatggccag tgtgccggcc tccgttatcg gggaagaagt ggctgatctc
     4981 agccaccgcg aaaatgacat caaaaacgcc attaacctga tgttctgggg aatataaatg
     5041 tcaggctccc ttatacacag ccagtctgca ggtcgaccat agtgactgga tatgttgtgt
     5101 tttacagtat tatgtagtct gttttttatg caaaatctaa tttaatatat tgatatttat
     5161 atcattttac gtttctcgtt cagctttctt gtacaaagtg gtcacctgca aaaagtgcca
     5221 gccaggacag aaatgcctcg acttcgctgc tacccaaggt tgccgggtga cgcacaccgt
     5281 ggaaacggat gaaggcacga acccagtgga cataagcctg ttcggttcgt aagctgtaat
     5341 gcaagtagcg tatgcgctca cgcaactggt ccagaacctt gaccgaacgc agcggtggta
     5401 acggcgcagt ggcggttttc atggcttgtt atgactgttt ttttggggta cagtctatgc
     5461 ctcgggcatc caagcagcaa gcgcgttacg ccgtgggtcg atgtttgatg ttatggagca
     5521 gcaacgatgt tacgcagcag ggcagtcgcc ctaaaacaaa gttaaacatt atgagggaag
     5581 cggtgatcgc cgaagtatcg actcaactat cagaggtagt tggcgtcatc gagcgccatc
     5641 tcgaaccgac gttgctggcc gtacatttgt acggctccgc agtggatggc ggcctgaagc
     5701 cacacagtga tattgatttg ctggttacgg tgaccgtaag gcttgatgaa acaacgcggc
     5761 gagctttgat caacgacctt ttggaaactt cggcttcccc tggagagagc gagattctcc
     5821 gcgctgtaga agtcaccatt gttgtgcacg acgacatcat tccgtggcgt tatccagcta
     5881 agcgcgaact gcaatttgga gaatggcagc gcaatgacat tcttgcaggt atcttcgagc
     5941 cagccacgat cgacattgat ctggctatct tgctgacaaa agcaagagaa catagcgttg
     6001 ccttggtagg tccagcggcg gaggaactct ttgatccggt tcctgaacag gatctatttg
     6061 aggcgctaaa tgaaacctta acgctatgga actcgccgcc cgactgggct ggcgatgagc
     6121 gaaatgtagt gcttacgttg tcccgcattt ggtacagcgc agtaaccggc aaaatcgcgc
     6181 cgaaggatgt cgctgccgac tgggcaatgg agcgcctgcc ggcccagtat cagcccgtca
     6241 tacttgaagc tagacaggct tatcttggac aagaagaaga tcgcttggcc tcgcgcgcag
     6301 atcagttgga agaatttgtc cactacgtga aaggcgagat caccaaggta gtcggcaaat
     6361 aaccctcgag c
//
