LOCUS       sequence-182143-        6224 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Transformation, Type: non-T-DNA, Plant Selection: none, Viral 
            Replication: ToLCV.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6224)
  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 6224)
  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_102
FEATURES             Location/Qualifiers
     source          1..6224
                     /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    1214..1338
                     /gene="mutant version of attR"
                     /label=attR1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        join(1363..1368,1369..1386,1387..1393)
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     CDS             1447..2127
                     /codon_start=1
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQAGRNLE
                     DPAY"
     CDS             2447..2752
                     /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     2631..2650
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     protein_bind    complement(2793..2917)
                     /gene="mutant version of attR"
                     /label=attR2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     terminator      complement(2954..3206)
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     CDS             5424..6215
                     /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 ccgcgtcgat taagttgact
      901 tggtcaatcg gtgtctctca actctctcta tgtatcggtg tattggagtc ctatatatat
      961 ggagactcta atggcataaa tgtaaatatg gaactttaaa tcaaaaccct tacgctccaa
     1021 aaagcggcca tccgtataat attaccggat ggccgcgatt ttttttttgg accccacaac
     1081 gcaataactg acaaagacat gcggaccaat cacctgacgc gctcaaagct taattgtttt
     1141 gtgttttcta tttaaacttg ggcaccaagt attgtagttc gtactacaag tttgtaccga
     1201 tcaaaagcag gtgacaagtt tgtacaaaaa agctgaacga gaaacgtaaa atgatataaa
     1261 tatcaatata ttaaattaga ttttgcataa aaaacagact acataatact gtaaaacaca
     1321 acatatccag tcactatggc ggccgcatta ggcaccccag gctttacact ttatgcttcc
     1381 ggctcgtata atgtgtggat tttgagttag gatccgtcga gattttcagg agctaaggaa
     1441 gctaaaatgg agaaaaaaat cactggatat accaccgttg atatatccca atggcatcgt
     1501 aaagaacatt ttgaggcatt tcagtcagtc gctcaatgta cctataacca gaccgttcag
     1561 ctggatatta cggccttttt aaagaccgta aagaaaaata agcacaagtt ttatccggcc
     1621 tttattcaca ttcttgcccg cctgatgaat gctcatccgg aattccgtat ggcaatgaaa
     1681 gacggtgagc tggtgatatg ggatagtgtt cacccttgtt acaccgtttt ccatgagcaa
     1741 actgaaacgt tttcatcgct ctggagtgaa taccacgacg atttccggca gtttctacac
     1801 atatattcgc aagatgtggc gtgttacggt gaaaacctgg cctatttccc taaagggttt
     1861 attgagaata tgtttttcgt ctcagccaat ccctgggtga gtttcaccag ttttgattta
     1921 aacgtggcca atatggacaa cttcttcgcc cccgttttca ccatgggcaa atattatacg
     1981 caaggcgaca aggtgctgat gccgctggcg attcaggttc atcatgccgt ttgtgatggc
     2041 ttccatgtcg gcagaatgct taatgaatta caacagtact gcgatgagtg gcaggcgggg
     2101 cgtaatctag aggatccggc ttactaaaag ccagataaca gtatgcgtat ttgcgcgctg
     2161 atttttgcgg tataagaata tatactgata tgtatacccg aagtatgtca aaaagaggta
     2221 tgctatgaag cagcgtatta cagtgacagt tgacagcgac agctatcagt tgctcaaggc
     2281 atatatgatg tcaatatctc cggtctggta agcacaacca tgcagaatga agcccgtcgt
     2341 ctgcgtgccg aacgctggaa agcggaaaat caggaaggga tggctgaggt cgcccggttt
     2401 attgaaatga acggctcttt tgctgacgag aacaggggct ggtgaaatgc agtttaaggt
     2461 ttacacctat aaaagagaga gccgttatcg tctgtttgtg gatgtacaga gtgatattat
     2521 tgacacgccc gggcgacgga tggtgatccc cctggccagt gcacgtctgc tgtcagataa
     2581 agtcccccgt gaactttacc cggtggtgca tatcggggat gaaagctggc gcatgatgac
     2641 caccgatatg gccagtgtgc cggtctccgt tatcggggaa gaagtggctg atctcagcca
     2701 ccgcgaaaat gacatcaaaa acgccattaa cctgatgttc tggggaatat aaatgtcagg
     2761 ctcccttata cacagccagt ctgcaggtcg accatagtga ctggatatgt tgtgttttac
     2821 agtattatgt agtctgtttt ttatgcaaaa tctaatttaa tatattgata tttatatcat
     2881 tttacgtttc tcgttcagct ttcttgtaca aagtggtcac ctgcaaaaag tgtttgatcg
     2941 ccggcggtac aaagtggtag taacatagat gacaccgcgc gcgataattt atcctagttt
     3001 gcgcgctata ttttgttttc tatcgcgtat taaatgtata attgcgggac tctaatcata
     3061 aaaacccatc tcataaataa cgtcatgcat tacatgttaa ttattacatg cttaacgtaa
     3121 ttcaacagaa attatatgat aatcatcgca agaccggcaa caggattcaa tcttaagaaa
     3181 ctttattgcc aaatgtttga acgatcgggg aaattcctaa taaatattaa attttattat
     3241 atgcgaactg tctacaaatg ttgtttgcgt caatgcatcc catagtacat aattcactgc
     3301 tctaattaca ttattcaaac taattacacc caaattatta agaaatttca acacttgtgt
     3361 tttaaatacc cttaagaaac gaccagtcgg aggctgtgag gtcatccaga ttcggtaggt
     3421 taggaaacat ttgtgtatcc ccaacacttt cctcaggttg tgattgaact gtatccggac
     3481 ggtgatgatg tcttcgttca tcacgaaggg ccggttttgg tgctctgtta tcttgaaata
     3541 taggggattt tgaatctccc agataaacac gccattctct gcttgagctg cagtgatgag
     3601 ttcccctgtg cgtgaatcca tggtcgtggc aggttattgc tatgaagtaa gaacagccac
     3661 acggtagatc aactcgtcga cgtctggtcc ccttcttggc tagcctgtgc tgtactttga
     3721 ttggtacctg agtagagtgg gccttcgagg gtgacgaagg tcgcattttt taaagcccag
     3781 ttttttagtg cagaattctt ttcctcatcc aagtattctt tatagctgga gttgggtcct
     3841 ggattgcaga ggaaggtagt gggaattccg cctttaattt gaactggctt cccgtacttt
     3901 gtatttgatt gccagtccct ttgggccccc atgaattctt taaagtgctt taggtagtgg
     3961 gggtctacgt catcaatgac gttgtaccag gcatcattac tgtaaacctt tgggctaagg
     4021 tctagatgac cacacaaata attatgtggt cccaatgatc tggcccacat agtcttcccc
     4081 gtcctactgt caccctcaat tactatactt atgggtctca gaggccgcgc agcgggactg
     4141 cagacgttct cggcagccca ctcttcaagt tcttctggaa cttggtcgaa agaagaagat
     4201 aaaaaaggtg aaacataaac atccattgga ggtgtaaaaa tcctatctaa attagaattt
     4261 aaattgtgaa attgtaaaac aaaatctttg ggagctttct ccctcaatat attaagggcc
     4321 tgagctttgg accctgaatt gattgcctcg gcatatgcgt cgttggcaga ttggcaacct
     4381 cctctagctg atcgtccatc gacttggaaa actccatgat caatgacgtc tccgtctttt
     4441 tccatgtagg atttgacatc gcttgagctc ttagctccct gaatgttcgg atggaaatgt
     4501 gctgacctgg ttggggatgt gaggtcgaag aatctattgt ttttgcactg gaactttcct
     4561 tcgaactgga tgagcacatg caagtgagga gtcccatctt cgtgaagctc tctgcaaatt
     4621 ctaataaatt ttttgaaagt gggtgtttgg agatttaata atttgggaaa gtgctcttct
     4681 ttagttagag agcacttggg ataagtgaga aaataatttt tggcatttat tttaaagcga
     4741 ttgggggctg ccatgttgac ttggtcaatc ggtgtctctc aactctctct atgtatcggt
     4801 gtattggagt cctatatata tggagactct aatggcataa atgtaaatat ggaactttaa
     4861 atcaaaaccc ttacgctcca aaaagcggcc atccgtataa tattaccgga tggccgcgat
     4921 tttttttttg gaccccacaa cgcaataact gacaaagaca tgcggaccaa tcacctgacg
     4981 cgctcaaagc ttaattgttt tgtgttttct atttaaactt gggcaccaag tattgtagtt
     5041 cgtacttggg gccctcgagt ttgtttaaac ccagccagga cagaaatgcc tcgacttcgc
     5101 tgctacccaa ggttgccggg tgacgcacac cgtggaaacg gatgaaggca cgaacccagt
     5161 ggacataagc ctgttcggtt cgtaagctgt aatgcaagta gcgtatgcgc tcacgcaact
     5221 ggtccagaac cttgaccgaa cgcagcggtg gtaacggcgc agtggcggtt ttcatggctt
     5281 gttatgactg tttttttggg gtacagtcta tgcctcgggc atccaagcag caagcgcgtt
     5341 acgccgtggg tcgatgtttg atgttatgga gcagcaacga tgttacgcag cagggcagtc
     5401 gccctaaaac aaagttaaac attatgaggg aagcggtgat cgccgaagta tcgactcaac
     5461 tatcagaggt agttggcgtc atcgagcgcc atctcgaacc gacgttgctg gccgtacatt
     5521 tgtacggctc cgcagtggat ggcggcctga agccacacag tgatattgat ttgctggtta
     5581 cggtgaccgt aaggcttgat gaaacaacgc ggcgagcttt gatcaacgac cttttggaaa
     5641 cttcggcttc ccctggagag agcgagattc tccgcgctgt agaagtcacc attgttgtgc
     5701 acgacgacat cattccgtgg cgttatccag ctaagcgcga actgcaattt ggagaatggc
     5761 agcgcaatga cattcttgca ggtatcttcg agccagccac gatcgacatt gatctggcta
     5821 tcttgctgac aaaagcaaga gaacatagcg ttgccttggt aggtccagcg gcggaggaac
     5881 tctttgatcc ggttcctgaa caggatctat ttgaggcgct aaatgaaacc ttaacgctat
     5941 ggaactcgcc gcccgactgg gctggcgatg agcgaaatgt agtgcttacg ttgtcccgca
     6001 tttggtacag cgcagtaacc ggcaaaatcg cgccgaagga tgtcgctgcc gactgggcaa
     6061 tggagcgcct gccggcccag tatcagcccg tcatacttga agctagacag gcttatcttg
     6121 gacaagaaga agatcgcttg gcctcgcgcg cagatcagtt ggaagaattt gtccactacg
     6181 tgaaaggcga gatcaccaag gtagtcggca aataaccctc gagc
//
