LOCUS       sequence-182149-        6879 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Transformation, Type: non-T-DNA, Plant Selection: PvUbi2:hpt II, 
            Viral Replication: none.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6879)
  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 6879)
  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_150
FEATURES             Location/Qualifiers
     source          1..6879
                     /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    876..1050
                     /label=CaMV poly(A) signal
                     /note="cauliflower mosaic virus polyadenylation signal"
     CDS             complement(1091..2116)
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRSQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     K"
     protein_bind    4007..4131
                     /gene="mutant version of attR"
                     /label=attR1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        join(4156..4161,4162..4179,4180..4186)
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     CDS             4240..4920
                     /codon_start=1
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQAGRNLE
                     DPAY"
     CDS             5240..5545
                     /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     5424..5443
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     protein_bind    complement(5586..5710)
                     /gene="mutant version of attR"
                     /label=attR2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             6079..6870
                     /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 atatgcaatt cgggggatct ggattttagt actggatttt
      901 ggttttagga attagaaatt ttattgatag aagtatttta caaatacaaa tacatactaa
      961 gggtttctta tatgctcaac acatgagcga aaccctatag gaaccctaat tcccttatct
     1021 gggaactact cacacattat tatggagaaa ctcgagcttg tcgatcgaca gatcccggtc
     1081 ggcatctact ctatttcttt gccctcggac gagtgctggg gcgtcggttt ccactatcgg
     1141 cgagtacttc tacacagcca tcggtccaga cggccgcgct tctgcgggcg atttgtgtac
     1201 gcccgacagt cccggctccg gatcggacga ttgcgtcgca tcgaccctgc gcccaagctg
     1261 catcatcgaa attgccgtca accaagctct gatagagttg gtcaagacca atgcggagca
     1321 tatacgcccg gagtcgtggc gatcctgcaa gctccggatg cctccgctcg aagtagcgcg
     1381 tctgctgctc catacaagcc aaccacggcc tccagaagaa gatgttggcg acctcgtatt
     1441 gggaatcccc gaacatcgcc tcgctccagt caatgaccgc tgttatgcgg ccattgtccg
     1501 tcaggacatt gttggagccg aaatccgcgt gcacgaggtg ccggacttcg gggcagtcct
     1561 cggcccaaag catcagctca tcgagagcct gcgcgacgga cgcactgacg gtgtcgtcca
     1621 tcacagtttg ccagtgatac acatggggat cagcaatcgc gcatatgaaa tcacgccatg
     1681 tagtgtattg accgattcct tgcggtccga atgggccgaa cccgctcgtc tggctaagat
     1741 cggccgcagc gatcgcatcc atagcctccg cgaccggttg tagaacagcg ggcagttcgg
     1801 tttcaggcag gtcttgcaac gtgacaccct gtgaacggcg ggagatgcaa taggtcaggc
     1861 tctcgctaaa ctccccaatg tcaagcactt ccggaatcgg gagcgcggcc gatgcaaagt
     1921 gccgataaac ataacgatct ttgtagaaac catcggcgca gctatttacc cgcaggacat
     1981 atccacgccc tcctacatcg aagctgaaag cacgagattc ttcgccctcc gagagctgca
     2041 tcaggtcgga cacgctgtcg aacttttcga tcagaaactt ctcgacagac gtcgcggtga
     2101 gttcaggctt tttcataagc ttctgcaaaa gagaaccaga caacagggta agtgcctagc
     2161 agtaaacaaa cagaactcat cacaagcaaa cagcaacatc atattcatac caacaggtca
     2221 tgtgtgttca tcacatcatt agtactaagc atgccatcat ccaagtatat caaagtaagg
     2281 gcaaagagca tgcatgatca tcaggtgcac aaaagaatca tcaaattgta gcagtacata
     2341 tcttcatcta tcatgcatat ctatccataa caggacgatg catgttgacc aggtaaaagc
     2401 tacaggatcc tataggaaca gcagcgtata tatcttcacc aatcttagca tcttaatcat
     2461 gtggcacatg cagtttcaat ttaagcacat gagctagttg attatgaggt accagagaat
     2521 catcaaactg atgtagcagc atatatcctc atctatcatg caatctaatc taatctaact
     2581 aaacaggaaa ggtgtgctat tcagttaaaa gctaccgcat catatacaaa cggcagcata
     2641 agaaaaagca taatcatctt aatcatgcaa caaacgcaga ttcataataa gcccaagagc
     2701 tagcttgtga tgatcttatt ctactctgat ctacagcaat cagataacga cctaaccttg
     2761 cacatggcaa caaaacaatc gatcggacga atcagttgtt tgttcctagc tagcaccatc
     2821 gaaccagata atagatgcac gtacagatcc cgaaaacgaa cccaaaaaca gggcagacct
     2881 agctgaacct aggcagcgac ccagcagatc gtgagaacga tctcatctac gaacagccta
     2941 gaagcaaccc cacgattccc ggacaaacga cctaaaatcc ccacaaatca catgagcatg
     3001 acaggataaa cagcggaacc gatcagatct acacgaaaac cccacctccc agccacccac
     3061 gatcaggaaa cacgcggatc tagcatgatt tcgtcaacgc ctcagcctag ttcctagcca
     3121 cagaccaagc agaaccacca aaccacgccg agcgaggaga tggggcaaga ggacggggga
     3181 gacgatcgcc gtaccttgaa gcgggggaag gatcgccgag ggtcgcgagg agagcaattt
     3241 ggatttggaa cccgggggtt gtgcgctccg aacgatgaga cgatgtgaga ttgtgggaag
     3301 aggcgcggag ggccctgtat ttatgggctg cgacgggggg aggagaggtg gggagggttg
     3361 gggaaggaat cccccacccg tgccgtgacg gttccgggcc gtgtgagagg agcccgctcg
     3421 tctccgccac gcaatttccg cgatcggagc ggagctttcg agaggcggct ggatggttgg
     3481 tggccgttag atttgtagac gccgttaacg cctcgcctcc accgggaaga gttttgagca
     3541 gccgcttatg acaatggctt aacgacgtta gacggagcgt tagtggcagg ccatgataga
     3601 atagacgtat tgcaatggga ttataattaa ataaataaga atataataag atatggcaag
     3661 tcggcactca tgacatggtc ttcgaaatga tagtgctcac tttcttagcc gagaaagttg
     3721 acgcgactga tttagaagtt aagatattat ttctctcttc ttttctttcc tcgtcatata
     3781 aggatgaaat aaactttaga gattgccggt agtgattttg gatttcggcg atcaggcttg
     3841 gtttgccggt ttcggacggt gtgccttagg ccacccgcag tgtatcttgt aatgttcaac
     3901 cgataagcaa gggtggggct caagcaagta gtaaacaact atgtcaaatg tcaccatggt
     3961 tatggtcttg tttagttggc ttcggcgcgc cgatcaaaag caggtgacaa gtttgtacaa
     4021 aaaagctgaa cgagaaacgt aaaatgatat aaatatcaat atattaaatt agattttgca
     4081 taaaaaacag actacataat actgtaaaac acaacatatc cagtcactat ggcggccgca
     4141 ttaggcaccc caggctttac actttatgct tccggctcgt ataatgtgtg gattttgagt
     4201 taggatccgt cgagattttc aggagctaag gaagctaaaa tggagaaaaa aatcactgga
     4261 tataccaccg ttgatatatc ccaatggcat cgtaaagaac attttgaggc atttcagtca
     4321 gtcgctcaat gtacctataa ccagaccgtt cagctggata ttacggcctt tttaaagacc
     4381 gtaaagaaaa ataagcacaa gttttatccg gcctttattc acattcttgc ccgcctgatg
     4441 aatgctcatc cggaattccg tatggcaatg aaagacggtg agctggtgat atgggatagt
     4501 gttcaccctt gttacaccgt tttccatgag caaactgaaa cgttttcatc gctctggagt
     4561 gaataccacg acgatttccg gcagtttcta cacatatatt cgcaagatgt ggcgtgttac
     4621 ggtgaaaacc tggcctattt ccctaaaggg tttattgaga atatgttttt cgtctcagcc
     4681 aatccctggg tgagtttcac cagttttgat ttaaacgtgg ccaatatgga caacttcttc
     4741 gcccccgttt tcaccatggg caaatattat acgcaaggcg acaaggtgct gatgccgctg
     4801 gcgattcagg ttcatcatgc cgtttgtgat ggcttccatg tcggcagaat gcttaatgaa
     4861 ttacaacagt actgcgatga gtggcaggcg gggcgtaatc tagaggatcc ggcttactaa
     4921 aagccagata acagtatgcg tatttgcgcg ctgatttttg cggtataaga atatatactg
     4981 atatgtatac cagaagtatg tcaaaaagag gtatgctatg aagcagcgta ttacagtgac
     5041 agttgacagc gacagctatc agttgctcaa ggcatatatg atgtcaatat ctccggtctg
     5101 gtaagcacaa ccatgcagaa tgaagcccgt cgtctgcgtg ccgaacgctg gaaagcggaa
     5161 aatcaggaag ggatggctga ggtcgcccgg tttattgaaa tgaacggctc ttttgctgac
     5221 gagaacaggg gctggtgaaa tgcagtttaa ggtttacacc tataaaagag agagccgtta
     5281 tcgtctgttt gtggatgtac agagtgatat tattgacacg cccgggcgac ggatggtgat
     5341 ccccctggcc agtgcacgtc tgctgtcaga taaagtcccc cgtgaacttt acccggtggt
     5401 gcatatcggg gatgaaagct ggcgcatgat gaccaccgat atggccagtg tgccggcctc
     5461 cgttatcggg gaagaagtgg ctgatctcag ccaccgcgaa aatgacatca aaaacgccat
     5521 taacctgatg ttctggggaa tataaatgtc aggctccctt atacacagcc agtctgcagg
     5581 tcgaccatag tgactggata tgttgtgttt tacagtatta tgtagtctgt tttttatgca
     5641 aaatctaatt taatatattg atatttatat cattttacgt ttctcgttca gctttcttgt
     5701 acaaagtggt cacctgcaaa aagtgccagc caggacagaa atgcctcgac ttcgctgcta
     5761 cccaaggttg ccgggtgacg cacaccgtgg aaacggatga aggcacgaac ccagtggaca
     5821 taagcctgtt cggttcgtaa gctgtaatgc aagtagcgta tgcgctcacg caactggtcc
     5881 agaaccttga ccgaacgcag cggtggtaac ggcgcagtgg cggttttcat ggcttgttat
     5941 gactgttttt ttggggtaca gtctatgcct cgggcatcca agcagcaagc gcgttacgcc
     6001 gtgggtcgat gtttgatgtt atggagcagc aacgatgtta cgcagcaggg cagtcgccct
     6061 aaaacaaagt taaacattat gagggaagcg gtgatcgccg aagtatcgac tcaactatca
     6121 gaggtagttg gcgtcatcga gcgccatctc gaaccgacgt tgctggccgt acatttgtac
     6181 ggctccgcag tggatggcgg cctgaagcca cacagtgata ttgatttgct ggttacggtg
     6241 accgtaaggc ttgatgaaac aacgcggcga gctttgatca acgacctttt ggaaacttcg
     6301 gcttcccctg gagagagcga gattctccgc gctgtagaag tcaccattgt tgtgcacgac
     6361 gacatcattc cgtggcgtta tccagctaag cgcgaactgc aatttggaga atggcagcgc
     6421 aatgacattc ttgcaggtat cttcgagcca gccacgatcg acattgatct ggctatcttg
     6481 ctgacaaaag caagagaaca tagcgttgcc ttggtaggtc cagcggcgga ggaactcttt
     6541 gatccggttc ctgaacagga tctatttgag gcgctaaatg aaaccttaac gctatggaac
     6601 tcgccgcccg actgggctgg cgatgagcga aatgtagtgc ttacgttgtc ccgcatttgg
     6661 tacagcgcag taaccggcaa aatcgcgccg aaggatgtcg ctgccgactg ggcaatggag
     6721 cgcctgccgg cccagtatca gcccgtcata cttgaagcta gacaggctta tcttggacaa
     6781 gaagaagatc gcttggcctc gcgcgcagat cagttggaag aatttgtcca ctacgtgaaa
     6841 ggcgagatca ccaaggtagt cggcaaataa ccctcgagc
//
