LOCUS       sequence-182147-        5294 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Transformation, Type: non-T-DNA, Plant Selection: 2x35S:bar, Viral 
            Replication: none.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5294)
  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 5294)
  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_130
FEATURES             Location/Qualifiers
     source          1..5294
                     /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    872..1046
                     /label=CaMV poly(A) signal
                     /note="cauliflower mosaic virus polyadenylation signal"
     CDS             complement(1053..1604)
                     /codon_start=1
                     /gene="Streptomyces hygroscopicus bar"
                     /product="phosphinothricin acetyltransferase"
                     /label=BlpR
                     /note="confers resistance to bialophos or phosphinothricin"
                     /translation="MSPERRPADIRRATEADMPAVCTIVNHYIETSTVNFRTEPQEPQE
                     WTDDLVRLRERYPWLVAEVDGEVAGIAYAGPWKARNAYDWTAESTVYVSPRHQRTGLGS
                     TLYTHLLKSLEAQGFKSVVAVIGLPNDPSVRMHEALGYAPRGMLRAAGFKHGNWHDVGF
                     WQLDFSLPVPPRPVLPVTEI"
     promoter        complement(1649..2325)
                     /label=CaMV 35S promoter (enhanced)
                     /note="cauliflower mosaic virus 35S promoter with a 
                     duplicated enhancer region"
     primer_bind     complement(1692..1712)
                     /label=35S promoter
                     /note="CaMV 35S promoter, forward primer"
     protein_bind    2422..2546
                     /gene="mutant version of attR"
                     /label=attR1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        join(2571..2576,2577..2594,2595..2601)
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     CDS             2655..3335
                     /codon_start=1
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQAGRNLE
                     DPAY"
     CDS             3655..3960
                     /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     3839..3858
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     protein_bind    complement(4001..4125)
                     /gene="mutant version of attR"
                     /label=attR2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             4494..5285
                     /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 ataattcggg ggatctggat tttagtactg gattttggtt
      901 ttaggaatta gaaattttat tgatagaagt attttacaaa tacaaataca tactaagggt
      961 ttcttatatg ctcaacacat gagcgaaacc ctataggaac cctaattccc ttatctggga
     1021 actactcaca cattattatg gagaaactcg agtcaaatct cggtgacggg caggaccgga
     1081 cggggcggta ccggcaggct gaagtccagc tgccagaaac ccacgtcatg ccagttcccg
     1141 tgcttgaagc cggccgcccg cagcatgccg cggggggcat atccgagcgc ctcgtgcatg
     1201 cgcacgctcg ggtcgttggg cagcccgatg acagcgacca cgctcttgaa gccctgtgcc
     1261 tccagggact tcagcagatg ggtgtagagc gtggagccca gtcccgtccg ctggtggcgg
     1321 gggctgacgt acacggtcga ctcggccgtc cagtcgtagg cgttgcgtgc cttccagggg
     1381 cccgcgtagg cgatgccggc gacctcgccg tccacctcgg cgacgagcca gggatagcgc
     1441 tcccgcagac ggacgaggtc gtccgtccac tcctgcggtt cctgcggctc ggtacggaag
     1501 ttgaccgtgc ttgtctcgat gtagtggttg acgatggtgc agaccgccgg catgtccgcc
     1561 tcggtggcac ggcggatgtc ggccgggcgt cgttctgggc tcatggtaga ctcgagagag
     1621 atagatttgt agagagagac tggtgatttc agcgtgtcct ctccaaatga aatgaacttc
     1681 cttatataga ggaagggtct tgcgaaggat agtgggattg tgcgtcatcc cttacgtcag
     1741 tggagatatc acatcaatcc acttgctttg aagacgtggt tggaacgtct tctttttcca
     1801 cgatgctcct cgtgggtggg ggtccatctt tgggaccact gtcggcagag gcatcttgaa
     1861 cgatagcctt tcctttatcg caatgatggc atttgtaggt gccaccttcc ttttctactg
     1921 tccttttgat gaagtgacag atagctgggc aatggaatcc gaggaggttt cccgatatta
     1981 ccctttgttg aaaagtctca atagcccttt ggtcttctga gactgtatct ttgatattct
     2041 tggagtagac gagagtgtcg tgctccacca tgttcacatc aatccacttg ctttgaagac
     2101 gtggttggaa cgtcttcttt ttccacgatg ctcctcgtgg gtgggggtcc atctttggga
     2161 ccactgtcgg cagaggcatc ttgaacgata gcctttcctt tatcgcaatg atggcatttg
     2221 taggtgccac cttccttttc tactgtcctt ttgatgaagt gacagatagc tgggcaatgg
     2281 aatccgagga ggtttcccga tattaccctt tgttgaaaag tctcaatagc cctttggtct
     2341 tctgagactg tatctttgat attcttggag tagacgagag tgtcgtgctc caccatgcgg
     2401 cgcgccgatc aaaagcaggt gacaagtttg tacaaaaaag ctgaacgaga aacgtaaaat
     2461 gatataaata tcaatatatt aaattagatt ttgcataaaa aacagactac ataatactgt
     2521 aaaacacaac atatccagtc actatggcgg ccgcattagg caccccaggc tttacacttt
     2581 atgcttccgg ctcgtataat gtgtggattt tgagttagga tccgtcgaga ttttcaggag
     2641 ctaaggaagc taaaatggag aaaaaaatca ctggatatac caccgttgat atatcccaat
     2701 ggcatcgtaa agaacatttt gaggcatttc agtcagtcgc tcaatgtacc tataaccaga
     2761 ccgttcagct ggatattacg gcctttttaa agaccgtaaa gaaaaataag cacaagtttt
     2821 atccggcctt tattcacatt cttgcccgcc tgatgaatgc tcatccggaa ttccgtatgg
     2881 caatgaaaga cggtgagctg gtgatatggg atagtgttca cccttgttac accgttttcc
     2941 atgagcaaac tgaaacgttt tcatcgctct ggagtgaata ccacgacgat ttccggcagt
     3001 ttctacacat atattcgcaa gatgtggcgt gttacggtga aaacctggcc tatttcccta
     3061 aagggtttat tgagaatatg tttttcgtct cagccaatcc ctgggtgagt ttcaccagtt
     3121 ttgatttaaa cgtggccaat atggacaact tcttcgcccc cgttttcacc atgggcaaat
     3181 attatacgca aggcgacaag gtgctgatgc cgctggcgat tcaggttcat catgccgttt
     3241 gtgatggctt ccatgtcggc agaatgctta atgaattaca acagtactgc gatgagtggc
     3301 aggcggggcg taatctagag gatccggctt actaaaagcc agataacagt atgcgtattt
     3361 gcgcgctgat ttttgcggta taagaatata tactgatatg tataccagaa gtatgtcaaa
     3421 aagaggtatg ctatgaagca gcgtattaca gtgacagttg acagcgacag ctatcagttg
     3481 ctcaaggcat atatgatgtc aatatctccg gtctggtaag cacaaccatg cagaatgaag
     3541 cccgtcgtct gcgtgccgaa cgctggaaag cggaaaatca ggaagggatg gctgaggtcg
     3601 cccggtttat tgaaatgaac ggctcttttg ctgacgagaa caggggctgg tgaaatgcag
     3661 tttaaggttt acacctataa aagagagagc cgttatcgtc tgtttgtgga tgtacagagt
     3721 gatattattg acacgcccgg gcgacggatg gtgatccccc tggccagtgc acgtctgctg
     3781 tcagataaag tcccccgtga actttacccg gtggtgcata tcggggatga aagctggcgc
     3841 atgatgacca ccgatatggc cagtgtgccg gcctccgtta tcggggaaga agtggctgat
     3901 ctcagccacc gcgaaaatga catcaaaaac gccattaacc tgatgttctg gggaatataa
     3961 atgtcaggct cccttataca cagccagtct gcaggtcgac catagtgact ggatatgttg
     4021 tgttttacag tattatgtag tctgtttttt atgcaaaatc taatttaata tattgatatt
     4081 tatatcattt tacgtttctc gttcagcttt cttgtacaaa gtggtcacct gcaaaaagtg
     4141 ccagccagga cagaaatgcc tcgacttcgc tgctacccaa ggttgccggg tgacgcacac
     4201 cgtggaaacg gatgaaggca cgaacccagt ggacataagc ctgttcggtt cgtaagctgt
     4261 aatgcaagta gcgtatgcgc tcacgcaact ggtccagaac cttgaccgaa cgcagcggtg
     4321 gtaacggcgc agtggcggtt ttcatggctt gttatgactg tttttttggg gtacagtcta
     4381 tgcctcgggc atccaagcag caagcgcgtt acgccgtggg tcgatgtttg atgttatgga
     4441 gcagcaacga tgttacgcag cagggcagtc gccctaaaac aaagttaaac attatgaggg
     4501 aagcggtgat cgccgaagta tcgactcaac tatcagaggt agttggcgtc atcgagcgcc
     4561 atctcgaacc gacgttgctg gccgtacatt tgtacggctc cgcagtggat ggcggcctga
     4621 agccacacag tgatattgat ttgctggtta cggtgaccgt aaggcttgat gaaacaacgc
     4681 ggcgagcttt gatcaacgac cttttggaaa cttcggcttc ccctggagag agcgagattc
     4741 tccgcgctgt agaagtcacc attgttgtgc acgacgacat cattccgtgg cgttatccag
     4801 ctaagcgcga actgcaattt ggagaatggc agcgcaatga cattcttgca ggtatcttcg
     4861 agccagccac gatcgacatt gatctggcta tcttgctgac aaaagcaaga gaacatagcg
     4921 ttgccttggt aggtccagcg gcggaggaac tctttgatcc ggttcctgaa caggatctat
     4981 ttgaggcgct aaatgaaacc ttaacgctat ggaactcgcc gcccgactgg gctggcgatg
     5041 agcgaaatgt agtgcttacg ttgtcccgca tttggtacag cgcagtaacc ggcaaaatcg
     5101 cgccgaagga tgtcgctgcc gactgggcaa tggagcgcct gccggcccag tatcagcccg
     5161 tcatacttga agctagacag gcttatcttg gacaagaaga agatcgcttg gcctcgcgcg
     5221 cagatcagtt ggaagaattt gtccactacg tgaaaggcga gatcaccaag gtagtcggca
     5281 aataaccctc gagc
//
