LOCUS       sequence-175639-        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      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(1087..2112)
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     K"
     CDS             complement(1087..2112)
                     /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(2179..2855)
                     /label=CaMV 35S promoter (enhanced)
                     /note="cauliflower mosaic virus 35S promoter with a 
                     duplicated enhancer region"
     primer_bind     complement(2222..2242)
                     /label=35S promoter
                     /note="CaMV 35S promoter, forward primer"
     protein_bind    2952..3076
                     /gene="mutant version of attR"
                     /label=attR1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        join(3101..3106,3107..3124,3125..3131)
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     CDS             3185..3865
                     /codon_start=1
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQAGRNLE
                     DPAY"
     CDS             4185..4490
                     /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     4369..4388
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     protein_bind    complement(4531..4655)
                     /gene="mutant version of attR"
                     /label=attR2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             5024..5815
                     /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 ccttctagtg
      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 tgagcattga
      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 agcttgtcga tcgacagatc ccggtcggca
     1081 tctactctat ttctttgccc tcggacgagt gctggggcgt cggtttccac tatcggcgag
     1141 tacttctaca cagccatcgg tccagacggc cgcgcttctg cgggcgattt gtgtacgccc
     1201 gacagtcccg gctccggatc ggacgattgc gtcgcatcga ccctgcgccc aagctgcatc
     1261 atcgaaattg ccgtcaacca agctctgata gagttggtca agaccaatgc ggagcatata
     1321 cgcccggagt cgtggcgatc ctgcaagctc cggatgcctc cgctcgaagt agcgcgtctg
     1381 ctgctccata caagccaacc acggcctcca gaagaagatg ttggcgacct cgtattggga
     1441 atccccgaac atcgcctcgc tccagtcaat gaccgctgtt atgcggccat tgtccgtcag
     1501 gacattgttg gagccgaaat ccgcgtgcac gaggtgccgg acttcggggc agtcctcggc
     1561 ccaaagcatc agctcatcga gagcctgcgc gacggacgca ctgacggtgt cgtccatcac
     1621 agtttgccag tgatacacat ggggatcagc aatcgcgcat atgaaatcac gccatgtagt
     1681 gtattgaccg attccttgcg gtccgaatgg gccgaacccg ctcgtctggc taagatcggc
     1741 cgcagcgatc gcatccatgg cctccgcgac cggctgcaga acagcgggca gttcggtttc
     1801 aggcaggtct tgcaacgtga caccctgtgc acggcgggag atgcaatagg tcaggctctc
     1861 gctgaattcc ccaatgtcaa gcacttccgg aatcgggagc gcggccgatg caaagtgccg
     1921 ataaacataa cgatctttgt agaaaccatc ggcgcagcta tttacccgca ggacatatcc
     1981 acgccctcct acatcgaagc tgaaagcacg agattcttcg ccctccgaga gctgcatcag
     2041 gtcggacacg ctgtcgaact tttcgatcag aaacttctcg acagacgtcg cggtgagttc
     2101 aggctttttc atatctcatt gccccccgga tctgcgaaag ctcgagagag atagatttgt
     2161 agagagagac tggtgatttc agcgtgtcct ctccaaatga aatgaacttc cttatataga
     2221 ggaagggtct tgcgaaggat agtgggattg tgcgtcatcc cttacgtcag tggagatatc
     2281 acatcaatcc acttgctttg aagacgtggt tggaacgtct tctttttcca cgatgctcct
     2341 cgtgggtggg ggtccatctt tgggaccact gtcggcagag gcatcttgaa cgatagcctt
     2401 tcctttatcg caatgatggc atttgtaggt gccaccttcc ttttctactg tccttttgat
     2461 gaagtgacag atagctgggc aatggaatcc gaggaggttt cccgatatta ccctttgttg
     2521 aaaagtctca atagcccttt ggtcttctga gactgtatct ttgatattct tggagtagac
     2581 gagagtgtcg tgctccacca tgttcacatc aatccacttg ctttgaagac gtggttggaa
     2641 cgtcttcttt ttccacgatg ctcctcgtgg gtgggggtcc atctttggga ccactgtcgg
     2701 cagaggcatc ttgaacgata gcctttcctt tatcgcaatg atggcatttg taggtgccac
     2761 cttccttttc tactgtcctt ttgatgaagt gacagatagc tgggcaatgg aatccgagga
     2821 ggtttcccga tattaccctt tgttgaaaag tctcaatagc cctttggtct tctgagactg
     2881 tatctttgat attcttggag tagacgagag tgtcgtgctc caccatgcgg cgcgccgatc
     2941 aaaagcaggt gacaagtttg tacaaaaaag ctgaacgaga aacgtaaaat gatataaata
     3001 tcaatatatt aaattagatt ttgcataaaa aacagactac ataatactgt aaaacacaac
     3061 atatccagtc actatggcgg ccgcattagg aaccccaggc tttacacttt atgcttccgg
     3121 ctcgtataat gtgtggattt tgagttagga tccgtcgaga ttttcaggag ctaaggaagc
     3181 taaaatggag aaaaaaatca ctggatatac caccgttgat atatcccaat ggcatcgtaa
     3241 agaacatttt gaggcatttc agtcagtcgc tcaatgtacc tataaccaga ccgttcagct
     3301 ggatattacg gcctttttaa agaccgtaaa gaaaaataag cacaagtttt atccggcctt
     3361 tattcacatt cttgcccgcc tgatgaatgc tcatccggaa ttccgtatgg caatgaaaga
     3421 cggtgagctg gtgatatggg atagtgttca cccttgttac accgttttcc atgagcaaac
     3481 tgaaacgttt tcatcgctct ggagtgaata ccacgacgat ttccggcagt ttctacacat
     3541 atattcgcaa gatgtggcgt gttacggtga aaacctggcc tatttcccta aagggtttat
     3601 tgagaatatg tttttcgtct cagccaatcc ctgggtgagt ttcaccagtt ttgatttaaa
     3661 cgtggccaat atggacaact tcttcgcccc cgttttcacc atgggcaaat attatacgca
     3721 aggcgacaag gtgctgatgc cgctggcgat tcaggttcat catgccgttt gtgatggctt
     3781 ccatgtcggc agaatgctta atgaattaca acagtactgc gatgagtggc aggcggggcg
     3841 taatctagag gatccggctt actaaaagcc agataacagt atgcgtattt gcgcgctgat
     3901 ttttgcggta taagaatata tactgatatg tatacccgaa gtatgtcaaa aagaggtatg
     3961 ctatgaagca gcgtattaca gtgacagttg acagcgacag ctatcagttg ctcaaggcat
     4021 atatgatgtc aatatctccg gtctggtaag cacaaccatg cagaatgaag cccgtcgtct
     4081 gcgtgccgaa cgctggaaag cggaaaatca ggaagggatg gctgaggtcg cccggtttat
     4141 tgaaatgaac ggctcttttg ctgacgagaa caggggctgg tgaaatgcag tttaaggttt
     4201 acacctataa aagagagagc cgttatcgtc tgtttgtgga tgtacagagt gatattattg
     4261 acacgcccgg gcgacggatg gtgatccccc tggccagtgc acgtctgctg tcagataaag
     4321 tcccccgtga actttacccg gtggtgcata tcggggatga aagctggcgc atgatgacca
     4381 ccgatatggc cagtgtgccg gcctccgtta tcggggaaga agtggctgat ctcagccacc
     4441 gcgaaaatga catcaaaaac gccattaacc tgatgttctg gggaatataa atgtcaggct
     4501 cccttataca cagccagtct gcaggtcgac catagtgact ggatatgttg tgttttacag
     4561 tattatgtag tctgtttttt atgcaaaatc taatttaata tattgatatt tatatcattt
     4621 tacgtttctc gttcagcttt cttgtacaaa gtggtcacct gcaaaaagtg ccagccagga
     4681 cagaaatgcc tcgacttcgc tgctacccaa ggttgccggg tgacgcacac cgtggaaacg
     4741 gatgaaggca cgaacccagt ggacataagc ctgttcggtt cgtaagctgt aatgcaagta
     4801 gcgtatgcgc tcacgcaact ggtccagaac cttgaccgaa cgcagcggtg gtaacggcgc
     4861 agtggcggtt ttcatggctt gttatgactg tttttttggg gtacagtcta tgcctcgggc
     4921 atccaagcag caagcgcgtt acgccgtggg tcgatgtttg atgttatgga gcagcaacga
     4981 tgttacgcag cagggcagtc gccctaaaac aaagttaaac atcatgaggg aagcggtgat
     5041 cgccgaagta tcgactcaac tatcagaggt agttggcgtc atcgagcgcc atctcgaacc
     5101 gacgttgctg gccgtacatt tgtacggctc cgcagtggat ggcggcctga agccacacag
     5161 tgatattgat ttgctggtta cggtgaccgt aaggcttgat gaaacaacgc ggcgagcttt
     5221 gatcaacgac cttttggaaa cttcggcttc ccctggagag agcgagattc tccgcgctgt
     5281 agaagtcacc attgttgtgc acgacgacat cattccgtgg cgttatccag ctaagcgcga
     5341 actgcaattt ggagaatggc agcgcaatga cattcttgca ggtatcttcg agccagccac
     5401 gatcgacatt gatctggcta tcttgctgac aaaagcaaga gaacatagcg ttgccttggt
     5461 aggtccagcg gcggaggaac tctttgatcc ggttcctgaa caggatctat ttgaggcgct
     5521 aaatgaaacc ttaacgctat ggaactcgcc gcccgactgg gctggcgatg agcgaaatgt
     5581 agtgcttacg ttgtcccgca tttggtacag cgcagtaacc ggcaaaatcg cgccgaagga
     5641 tgtcgctgcc gactgggcaa tggagcgcct gccggcccag tatcagcccg tcatacttga
     5701 agctagacag gcttatcttg gacaagaaga agatcgcttg gcctcgcgcg cagatcagtt
     5761 ggaagaattt gtccactacg tgaaaggcga gatcaccaag gtagtcggca aataaccctc
     5821 gagc
//
