LOCUS       sequence-375939-        5818 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Transformation, Type: non-T-DNA, Plant Selection: none, Viral 
            Replication: BeYDV.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5818)
  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 5818)
  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_101
FEATURES             Location/Qualifiers
     source          1..5818
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      53..641
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     542..561
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     795..812
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    1254..1378
                     /gene="mutant version of attR"
                     /label=attR1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        join(1403..1408,1409..1426,1427..1433)
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     CDS             1487..2167
                     /codon_start=1
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQAGRNLE
                     DPAY"
     CDS             2487..2792
                     /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     2671..2690
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     protein_bind    complement(2833..2957)
                     /gene="mutant version of attR"
                     /label=attR2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             4987..5778
                     /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 acgcgtcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc
       61 ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg
      121 tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag
      181 cgcagatacc aaatactgtt cttctagtgt agccgtagtt aggccaccac ttcaagaact
      241 ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg
      301 gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc
      361 ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg
      421 aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg
      481 cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag
      541 ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc
      601 gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct
      661 ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct gcgttatccc
      721 ctgattctgt ggataaccgt attaccgcct ttgagtgagc tgataccgct cgccgcagcc
      781 gaacgaccga gcgcagcgag tcagtgagcg aggaagcggg agagcgccca tatgcgcact
      841 cctcgcatgc ctcaaagttt aacgcgttag cagaaggcat gttgttgtga ctccgagggg
      901 ttgcctcaaa ctctatctta taaccggcgt ggaggcatgg aggcaggggt attttggtca
      961 ttttaataga tagtggaaaa tgacgtggaa tttacttaaa gacgaagtct ttgcgacaag
     1021 ggggggccca cgccgaattt aatattaccg gcgtggcccc cccttatcgc gagtgcttta
     1081 gcacgagcgg tccagattta aagtagaaaa tttcccgccc actagggtta aaggtgttca
     1141 cactataaaa gcatatacga tgtgatggta tttgatggag cgtatattgt atcaggtatt
     1201 tccgttggat acgaattatt cgtacgaccc tcggtaccga tcaaaagcag gtgacaagtt
     1261 tgtacaaaaa agctgaacga gaaacgtaaa atgatataaa tatcaatata ttaaattaga
     1321 ttttgcataa aaaacagact acataatact gtaaaacaca acatatccag tcactatggc
     1381 ggccgcatta ggcaccccag gctttacact ttatgcttcc ggctcgtata atgtgtggat
     1441 tttgagttag gatccgtcga gattttcagg agctaaggaa gctaaaatgg agaaaaaaat
     1501 cactggatat accaccgttg atatatccca atggcatcgt aaagaacatt ttgaggcatt
     1561 tcagtcagtc gctcaatgta cctataacca gaccgttcag ctggatatta cggccttttt
     1621 aaagaccgta aagaaaaata agcacaagtt ttatccggcc tttattcaca ttcttgcccg
     1681 cctgatgaat gctcatccgg aattccgtat ggcaatgaaa gacggtgagc tggtgatatg
     1741 ggatagtgtt cacccttgtt acaccgtttt ccatgagcaa actgaaacgt tttcatcgct
     1801 ctggagtgaa taccacgacg atttccggca gtttctacac atatattcgc aagatgtggc
     1861 gtgttacggt gaaaacctgg cctatttccc taaagggttt attgagaata tgtttttcgt
     1921 ctcagccaat ccctgggtga gtttcaccag ttttgattta aacgtggcca atatggacaa
     1981 cttcttcgcc cccgttttca ccatgggcaa atattatacg caaggcgaca aggtgctgat
     2041 gccgctggcg attcaggttc atcatgccgt ttgtgatggc ttccatgtcg gcagaatgct
     2101 taatgaatta caacagtact gcgatgagtg gcaggcgggg cgtaatctag aggatccggc
     2161 ttactaaaag ccagataaca gtatgcgtat ttgcgcgctg atttttgcgg tataagaata
     2221 tatactgata tgtatacccg aagtatgtca aaaagaggta tgctatgaag cagcgtatta
     2281 cagtgacagt tgacagcgac agctatcagt tgctcaaggc atatatgatg tcaatatctc
     2341 cggtctggta agcacaacca tgcagaatga agcccgtcgt ctgcgtgccg aacgctggaa
     2401 agcggaaaat caggaaggga tggctgaggt cgcccggttt attgaaatga acggctcttt
     2461 tgctgacgag aacaggggct ggtgaaatgc agtttaaggt ttacacctat aaaagagaga
     2521 gccgttatcg tctgtttgtg gatgtacaga gtgatattat tgacacgccc gggcgacgga
     2581 tggtgatccc cctggccagt gcacgtctgc tgtcagataa agtcccccgt gaactttacc
     2641 cggtggtgca tatcggggat gaaagctggc gcatgatgac caccgatatg gccagtgtgc
     2701 cggtctccgt tatcggggaa gaagtggctg atctcagcca ccgcgaaaat gacatcaaaa
     2761 acgccattaa cctgatgttc tggggaatat aaatgtcagg ctcccttata cacagccagt
     2821 ctgcaggtcg accatagtga ctggatatgt tgtgttttac agtattatgt agtctgtttt
     2881 ttatgcaaaa tctaatttaa tatattgata tttatatcat tttacgtttc tcgttcagct
     2941 ttcttgtaca aagtggtcac ctgcaaaaag tgtttgatcg ccggcggtac cgagtgtact
     3001 tcaagtcagt gggaaatcaa taaaatgatt attttatgaa tatatttcat tgtgcaagta
     3061 gatagaaatt acatatgtta cataacacac gaaataaaca aaaaaagaca atccaaaaac
     3121 aaacacccca aaaaaaataa tcactttaga taaactcgta tgaggagagg cacgttcagt
     3181 gactcgacga ttcccgagca aaaaaagtct ccccgtcaca catgtagtgg gtgacgcaat
     3241 tatctttaaa gtaatccttc tgttgacttg tcattgataa catccagtct tcgtcaggat
     3301 tgcaaagaat tatagaaggg atcccacctt ttattttctt cttttttcca tatttagggt
     3361 tgacagtgaa atcagactgg caacctatta attgcttcca caatgggacg aacttgaagg
     3421 ggatgtcgtc gatgatatta taggtggcgt gttcatcgta gttggtgaaa tcgatggtac
     3481 cgttccaata gttgtgtcgt ccgagacttc tagcccaggt ggtctttccg gtacgagttg
     3541 gtccgcagat gtagaggctg gggtgtcgga ttccattcct tccattgtcc ttgttaaatc
     3601 ggccatccat tcaaggtcag attgagcttg ttggtatgag acaggatgta tgtaagtata
     3661 agcgtctatg cttacatggt atagatgggt ttccctccag gagtgtagat cttcgtggca
     3721 gcgaagatct gattctgtga agggcgacac atacggttca ggttgtggag ggaataattt
     3781 gttggctgaa tattccagcc attgaagctt tgttgcccat tcatgaggga attcttcctt
     3841 gatcatgtca agatattcct ccttagacgt tgcagtctgg ataatagttc tccatcgtgc
     3901 gtcagatttg cgaggagaaa ccttatgatc tcggaaatct cctctggttt taatatctcc
     3961 gtcctttgat atgtaatcaa ggacttgttt agagtttcta gctggctgga tattagggtg
     4021 atttccttca aaatcgaaaa aagaaggatc cctaatacaa ggttttttat caagctggag
     4081 aagagcatga tagtgggtag tgccatcttg atgaagctca gaagcaacac caaggaagaa
     4141 aataagaaaa ggtgtgagtt tctcccagag aaactggaat aaatcatctc tttgagatga
     4201 gcacttggga taggtaagga aaacatattt agattggagt ctgaagttct tactagcaga
     4261 aggcatgttg ttgtgactcc gaggggttgc ctcaaactct atcttataac cggcgtggag
     4321 gcatggaggc aggggtattt tggtcatttt aatagatagt ggaaaatgac gtggaattta
     4381 cttaaagacg aagtctttgc gacaaggggg ggcccacgcc gaatttaata ttaccggcgt
     4441 ggccccccct tatcgcgagt gctttagcac gagcggtcca gatttaaagt agaaaatttc
     4501 ccgcccacta gggttaaagg tgttcacact ataaaagcat atacgatgtg atggtatttg
     4561 atggagcgta tattgtatca ggtatttccg ttggatacga attattcgta cgaccctcat
     4621 agtttgttta aacccagcca ggacagaaat gcctcgactt cgctgctacc caaggttgcc
     4681 gggtgacgca caccgtggaa acggatgaag gcacgaaccc agtggacata agcctgttcg
     4741 gttcgtaagc tgtaatgcaa gtagcgtatg cgctcacgca actggtccag aaccttgacc
     4801 gaacgcagcg gtggtaacgg cgcagtggcg gttttcatgg cttgttatga ctgttttttt
     4861 ggggtacagt ctatgcctcg ggcatccaag cagcaagcgc gttacgccgt gggtcgatgt
     4921 ttgatgttat ggagcagcaa cgatgttacg cagcagggca gtcgccctaa aacaaagtta
     4981 aacattatga gggaagcggt gatcgccgaa gtatcgactc aactatcaga ggtagttggc
     5041 gtcatcgagc gccatctcga accgacgttg ctggccgtac atttgtacgg ctccgcagtg
     5101 gatggcggcc tgaagccaca cagtgatatt gatttgctgg ttacggtgac cgtaaggctt
     5161 gatgaaacaa cgcggcgagc tttgatcaac gaccttttgg aaacttcggc ttcccctgga
     5221 gagagcgaga ttctccgcgc tgtagaagtc accattgttg tgcacgacga catcattccg
     5281 tggcgttatc cagctaagcg cgaactgcaa tttggagaat ggcagcgcaa tgacattctt
     5341 gcaggtatct tcgagccagc cacgatcgac attgatctgg ctatcttgct gacaaaagca
     5401 agagaacata gcgttgcctt ggtaggtcca gcggcggagg aactctttga tccggttcct
     5461 gaacaggatc tatttgaggc gctaaatgaa accttaacgc tatggaactc gccgcccgac
     5521 tgggctggcg atgagcgaaa tgtagtgctt acgttgtccc gcatttggta cagcgcagta
     5581 accggcaaaa tcgcgccgaa ggatgtcgct gccgactggg caatggagcg cctgccggcc
     5641 cagtatcagc ccgtcatact tgaagctaga caggcttatc ttggacaaga agaagatcgc
     5701 ttggcctcgc gcgcagatca gttggaagaa tttgtccact acgtgaaagg cgagatcacc
     5761 aaggtagtcg gcaaataacc ctcgagccac ccatgaccaa aatcccttaa cgtgagtt
//
