LOCUS       sequence-208969-        5824 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Transformation, Type: non-T-DNA, Plant Selection: 2x35S:hpt II, 
            Viral Replication: none.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5824)
  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 5824)
  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_110
FEATURES             Location/Qualifiers
     source          1..5824
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      86..674
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     575..594
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     828..845
                     /label=L4440
                     /note="L4440 vector, forward primer"
     polyA_signal    874..1048
                     /label=CaMV poly(A) signal
                     /note="cauliflower mosaic virus polyadenylation signal"
     CDS             complement(1089..2114)
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     K"
     CDS             complement(1089..2114)
                     /codon_start=1
                     /gene="aph(4)-Ia"
                     /product="aminoglycoside phosphotransferase from E. coli"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     K"
     promoter        complement(2181..2857)
                     /label=CaMV 35S promoter (enhanced)
                     /note="cauliflower mosaic virus 35S promoter with a 
                     duplicated enhancer region"
     primer_bind     complement(2224..2244)
                     /label=35S promoter
                     /note="CaMV 35S promoter, forward primer"
     protein_bind    2954..3078
                     /gene="mutant version of attR"
                     /label=attR1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        join(3103..3108,3109..3126,3127..3133)
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     CDS             3187..3867
                     /codon_start=1
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQAGRNLE
                     DPAY"
     CDS             4187..4492
                     /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     4371..4390
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     protein_bind    complement(4533..4657)
                     /gene="mutant version of attR"
                     /label=attR2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             5026..5817
                     /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 gccacccatg accaaaatcc cttaacgtga gttacgcgtc gttccactga gcgtcagacc
       61 ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct
      121 tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa
      181 ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag
      241 tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc
      301 tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg
      361 actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca
      421 cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat
      481 gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg
      541 tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc
      601 ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc
      661 ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc
      721 cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg
      781 cctttgagtg agctgatacc gctcgccgca gccgaacgac cgagcgcagc gagtcagtga
      841 gcgaggaagc gggagagcgc ccataattcg ggggatctgg attttagtac tggattttgg
      901 ttttaggaat tagaaatttt attgatagaa gtattttaca aatacaaata catactaagg
      961 gtttcttata tgctcaacac atgagcgaaa ccctatagga accctaattc ccttatctgg
     1021 gaactactca cacattatta tggagaaact cgagcttgtc gatcgacaga tcccggtcgg
     1081 catctactct atttctttgc cctcggacga gtgctggggc gtcggtttcc actatcggcg
     1141 agtacttcta cacagccatc ggtccagacg gccgcgcttc tgcgggcgat ttgtgtacgc
     1201 ccgacagtcc cggctccgga tcggacgatt gcgtcgcatc gaccctgcgc ccaagctgca
     1261 tcatcgaaat tgccgtcaac caagctctga tagagttggt caagaccaat gcggagcata
     1321 tacgcccgga gtcgtggcga tcctgcaagc tccggatgcc tccgctcgaa gtagcgcgtc
     1381 tgctgctcca tacaagccaa ccacggcctc cagaagaaga tgttggcgac ctcgtattgg
     1441 gaatccccga acatcgcctc gctccagtca atgaccgctg ttatgcggcc attgtccgtc
     1501 aggacattgt tggagccgaa atccgcgtgc acgaggtgcc ggacttcggg gcagtcctcg
     1561 gcccaaagca tcagctcatc gagagcctgc gcgacggacg cactgacggt gtcgtccatc
     1621 acagtttgcc agtgatacac atggggatca gcaatcgcgc atatgaaatc acgccatgta
     1681 gtgtattgac cgattccttg cggtccgaat gggccgaacc cgctcgtctg gctaagatcg
     1741 gccgcagcga tcgcatccat ggcctccgcg accggctgca gaacagcggg cagttcggtt
     1801 tcaggcaggt cttgcaacgt gacaccctgt gcacggcggg agatgcaata ggtcaggctc
     1861 tcgctgaatt ccccaatgtc aagcacttcc ggaatcggga gcgcggccga tgcaaagtgc
     1921 cgataaacat aacgatcttt gtagaaacca tcggcgcagc tatttacccg caggacatat
     1981 ccacgccctc ctacatcgaa gctgaaagca cgagattctt cgccctccga gagctgcatc
     2041 aggtcggaca cgctgtcgaa cttttcgatc agaaacttct cgacagacgt cgcggtgagt
     2101 tcaggctttt tcatatctca ttgccccccg gatctgcgaa agctcgagag agatagattt
     2161 gtagagagag actggtgatt tcagcgtgtc ctctccaaat gaaatgaact tccttatata
     2221 gaggaagggt cttgcgaagg atagtgggat tgtgcgtcat cccttacgtc agtggagata
     2281 tcacatcaat ccacttgctt tgaagacgtg gttggaacgt cttctttttc cacgatgctc
     2341 ctcgtgggtg ggggtccatc tttgggacca ctgtcggcag aggcatcttg aacgatagcc
     2401 tttcctttat cgcaatgatg gcatttgtag gtgccacctt ccttttctac tgtccttttg
     2461 atgaagtgac agatagctgg gcaatggaat ccgaggaggt ttcccgatat taccctttgt
     2521 tgaaaagtct caatagccct ttggtcttct gagactgtat ctttgatatt cttggagtag
     2581 acgagagtgt cgtgctccac catgttcaca tcaatccact tgctttgaag acgtggttgg
     2641 aacgtcttct ttttccacga tgctcctcgt gggtgggggt ccatctttgg gaccactgtc
     2701 ggcagaggca tcttgaacga tagcctttcc tttatcgcaa tgatggcatt tgtaggtgcc
     2761 accttccttt tctactgtcc ttttgatgaa gtgacagata gctgggcaat ggaatccgag
     2821 gaggtttccc gatattaccc tttgttgaaa agtctcaata gccctttggt cttctgagac
     2881 tgtatctttg atattcttgg agtagacgag agtgtcgtgc tccaccatgc ggcgcgccga
     2941 tcaaaagcag gtgacaagtt tgtacaaaaa agctgaacga gaaacgtaaa atgatataaa
     3001 tatcaatata ttaaattaga ttttgcataa aaaacagact acataatact gtaaaacaca
     3061 acatatccag tcactatggc ggccgcatta ggcaccccag gctttacact ttatgcttcc
     3121 ggctcgtata atgtgtggat tttgagttag gatccgtcga gattttcagg agctaaggaa
     3181 gctaaaatgg agaaaaaaat cactggatat accaccgttg atatatccca atggcatcgt
     3241 aaagaacatt ttgaggcatt tcagtcagtc gctcaatgta cctataacca gaccgttcag
     3301 ctggatatta cggccttttt aaagaccgta aagaaaaata agcacaagtt ttatccggcc
     3361 tttattcaca ttcttgcccg cctgatgaat gctcatccgg aattccgtat ggcaatgaaa
     3421 gacggtgagc tggtgatatg ggatagtgtt cacccttgtt acaccgtttt ccatgagcaa
     3481 actgaaacgt tttcatcgct ctggagtgaa taccacgacg atttccggca gtttctacac
     3541 atatattcgc aagatgtggc gtgttacggt gaaaacctgg cctatttccc taaagggttt
     3601 attgagaata tgtttttcgt ctcagccaat ccctgggtga gtttcaccag ttttgattta
     3661 aacgtggcca atatggacaa cttcttcgcc cccgttttca ccatgggcaa atattatacg
     3721 caaggcgaca aggtgctgat gccgctggcg attcaggttc atcatgccgt ttgtgatggc
     3781 ttccatgtcg gcagaatgct taatgaatta caacagtact gcgatgagtg gcaggcgggg
     3841 cgtaatctag aggatccggc ttactaaaag ccagataaca gtatgcgtat ttgcgcgctg
     3901 atttttgcgg tataagaata tatactgata tgtataccag aagtatgtca aaaagaggta
     3961 tgctatgaag cagcgtatta cagtgacagt tgacagcgac agctatcagt tgctcaaggc
     4021 atatatgatg tcaatatctc cggtctggta agcacaacca tgcagaatga agcccgtcgt
     4081 ctgcgtgccg aacgctggaa agcggaaaat caggaaggga tggctgaggt cgcccggttt
     4141 attgaaatga acggctcttt tgctgacgag aacaggggct ggtgaaatgc agtttaaggt
     4201 ttacacctat aaaagagaga gccgttatcg tctgtttgtg gatgtacaga gtgatattat
     4261 tgacacgccc gggcgacgga tggtgatccc cctggccagt gcacgtctgc tgtcagataa
     4321 agtcccccgt gaactttacc cggtggtgca tatcggggat gaaagctggc gcatgatgac
     4381 caccgatatg gccagtgtgc cggcctccgt tatcggggaa gaagtggctg atctcagcca
     4441 ccgcgaaaat gacatcaaaa acgccattaa cctgatgttc tggggaatat aaatgtcagg
     4501 ctcccttata cacagccagt ctgcaggtcg accatagtga ctggatatgt tgtgttttac
     4561 agtattatgt agtctgtttt ttatgcaaaa tctaatttaa tatattgata tttatatcat
     4621 tttacgtttc tcgttcagct ttcttgtaca aagtggtcac ctgcaaaaag tgccagccag
     4681 gacagaaatg cctcgacttc gctgctaccc aaggttgccg ggtgacgcac accgtggaaa
     4741 cggatgaagg cacgaaccca gtggacataa gcctgttcgg ttcgtaagct gtaatgcaag
     4801 tagcgtatgc gctcacgcaa ctggtccaga accttgaccg aacgcagcgg tggtaacggc
     4861 gcagtggcgg ttttcatggc ttgttatgac tgtttttttg gggtacagtc tatgcctcgg
     4921 gcatccaagc agcaagcgcg ttacgccgtg ggtcgatgtt tgatgttatg gagcagcaac
     4981 gatgttacgc agcagggcag tcgccctaaa acaaagttaa acattatgag ggaagcggtg
     5041 atcgccgaag tatcgactca actatcagag gtagttggcg tcatcgagcg ccatctcgaa
     5101 ccgacgttgc tggccgtaca tttgtacggc tccgcagtgg atggcggcct gaagccacac
     5161 agtgatattg atttgctggt tacggtgacc gtaaggcttg atgaaacaac gcggcgagct
     5221 ttgatcaacg accttttgga aacttcggct tcccctggag agagcgagat tctccgcgct
     5281 gtagaagtca ccattgttgt gcacgacgac atcattccgt ggcgttatcc agctaagcgc
     5341 gaactgcaat ttggagaatg gcagcgcaat gacattcttg caggtatctt cgagccagcc
     5401 acgatcgaca ttgatctggc tatcttgctg acaaaagcaa gagaacatag cgttgccttg
     5461 gtaggtccag cggcggagga actctttgat ccggttcctg aacaggatct atttgaggcg
     5521 ctaaatgaaa ccttaacgct atggaactcg ccgcccgact gggctggcga tgagcgaaat
     5581 gtagtgctta cgttgtcccg catttggtac agcgcagtaa ccggcaaaat cgcgccgaag
     5641 gatgtcgctg ccgactgggc aatggagcgc ctgccggccc agtatcagcc cgtcatactt
     5701 gaagctagac aggcttatct tggacaagaa gaagatcgct tggcctcgcg cgcagatcag
     5761 ttggaagaat ttgtccacta cgtgaaaggc gagatcacca aggtagtcgg caaataaccc
     5821 tcga
//
