LOCUS       sequence-182140-        3777 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Transformation, Type: non-T-DNA, Plant Selection: none, Viral 
            Replication: none.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3777)
  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 3777)
  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_100
FEATURES             Location/Qualifiers
     source          1..3777
                     /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"
     protein_bind    905..1029
                     /gene="mutant version of attR"
                     /label=attR1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        join(1054..1059,1060..1077,1078..1084)
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     CDS             1138..1818
                     /codon_start=1
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQAGRNLE
                     DPAY"
     CDS             2138..2443
                     /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     2322..2341
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     protein_bind    complement(2484..2608)
                     /gene="mutant version of attR"
                     /label=attR2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             2977..3768
                     /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 atatgcgcac tcctcgcatg cggcgcgccg atcaaaagca
      901 ggtgacaagt ttgtacaaaa aagctgaacg agaaacgtaa aatgatataa atatcaatat
      961 attaaattag attttgcata aaaaacagac tacataatac tgtaaaacac aacatatcca
     1021 gtcactatgg cggccgcatt aggcacccca ggctttacac tttatgcttc cggctcgtat
     1081 aatgtgtgga ttttgagtta ggatccgtcg agattttcag gagctaagga agctaaaatg
     1141 gagaaaaaaa tcactggata taccaccgtt gatatatccc aatggcatcg taaagaacat
     1201 tttgaggcat ttcagtcagt cgctcaatgt acctataacc agaccgttca gctggatatt
     1261 acggcctttt taaagaccgt aaagaaaaat aagcacaagt tttatccggc ctttattcac
     1321 attcttgccc gcctgatgaa tgctcatccg gaattccgta tggcaatgaa agacggtgag
     1381 ctggtgatat gggatagtgt tcacccttgt tacaccgttt tccatgagca aactgaaacg
     1441 ttttcatcgc tctggagtga ataccacgac gatttccggc agtttctaca catatattcg
     1501 caagatgtgg cgtgttacgg tgaaaacctg gcctatttcc ctaaagggtt tattgagaat
     1561 atgtttttcg tctcagccaa tccctgggtg agtttcacca gttttgattt aaacgtggcc
     1621 aatatggaca acttcttcgc ccccgttttc accatgggca aatattatac gcaaggcgac
     1681 aaggtgctga tgccgctggc gattcaggtt catcatgccg tttgtgatgg cttccatgtc
     1741 ggcagaatgc ttaatgaatt acaacagtac tgcgatgagt ggcaggcggg gcgtaatcta
     1801 gaggatccgg cttactaaaa gccagataac agtatgcgta tttgcgcgct gatttttgcg
     1861 gtataagaat atatactgat atgtatacca gaagtatgtc aaaaagaggt atgctatgaa
     1921 gcagcgtatt acagtgacag ttgacagcga cagctatcag ttgctcaagg catatatgat
     1981 gtcaatatct ccggtctggt aagcacaacc atgcagaatg aagcccgtcg tctgcgtgcc
     2041 gaacgctgga aagcggaaaa tcaggaaggg atggctgagg tcgcccggtt tattgaaatg
     2101 aacggctctt ttgctgacga gaacaggggc tggtgaaatg cagtttaagg tttacaccta
     2161 taaaagagag agccgttatc gtctgtttgt ggatgtacag agtgatatta ttgacacgcc
     2221 cgggcgacgg atggtgatcc ccctggccag tgcacgtctg ctgtcagata aagtcccccg
     2281 tgaactttac ccggtggtgc atatcgggga tgaaagctgg cgcatgatga ccaccgatat
     2341 ggccagtgtg ccggcctccg ttatcgggga agaagtggct gatctcagcc accgcgaaaa
     2401 tgacatcaaa aacgccatta acctgatgtt ctggggaata taaatgtcag gctcccttat
     2461 acacagccag tctgcaggtc gaccatagtg actggatatg ttgtgtttta cagtattatg
     2521 tagtctgttt tttatgcaaa atctaattta atatattgat atttatatca ttttacgttt
     2581 ctcgttcagc tttcttgtac aaagtggtca cctgcaaaaa gtgccagcca ggacagaaat
     2641 gcctcgactt cgctgctacc caaggttgcc gggtgacgca caccgtggaa acggatgaag
     2701 gcacgaaccc agtggacata agcctgttcg gttcgtaagc tgtaatgcaa gtagcgtatg
     2761 cgctcacgca actggtccag aaccttgacc gaacgcagcg gtggtaacgg cgcagtggcg
     2821 gttttcatgg cttgttatga ctgttttttt ggggtacagt ctatgcctcg ggcatccaag
     2881 cagcaagcgc gttacgccgt gggtcgatgt ttgatgttat ggagcagcaa cgatgttacg
     2941 cagcagggca gtcgccctaa aacaaagtta aacattatga gggaagcggt gatcgccgaa
     3001 gtatcgactc aactatcaga ggtagttggc gtcatcgagc gccatctcga accgacgttg
     3061 ctggccgtac atttgtacgg ctccgcagtg gatggcggcc tgaagccaca cagtgatatt
     3121 gatttgctgg ttacggtgac cgtaaggctt gatgaaacaa cgcggcgagc tttgatcaac
     3181 gaccttttgg aaacttcggc ttcccctgga gagagcgaga ttctccgcgc tgtagaagtc
     3241 accattgttg tgcacgacga catcattccg tggcgttatc cagctaagcg cgaactgcaa
     3301 tttggagaat ggcagcgcaa tgacattctt gcaggtatct tcgagccagc cacgatcgac
     3361 attgatctgg ctatcttgct gacaaaagca agagaacata gcgttgcctt ggtaggtcca
     3421 gcggcggagg aactctttga tccggttcct gaacaggatc tatttgaggc gctaaatgaa
     3481 accttaacgc tatggaactc gccgcccgac tgggctggcg atgagcgaaa tgtagtgctt
     3541 acgttgtccc gcatttggta cagcgcagta accggcaaaa tcgcgccgaa ggatgtcgct
     3601 gccgactggg caatggagcg cctgccggcc cagtatcagc ccgtcatact tgaagctaga
     3661 caggcttatc ttggacaaga agaagatcgc ttggcctcgc gcgcagatca gttggaagaa
     3721 tttgtccact acgtgaaagg cgagatcacc aaggtagtcg gcaaataacc ctcgagc
//
