LOCUS       sequence-331269-       10053 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Wheat transformation binary vector pGGG confers hygromycin 
            resistance in wheat and contains the GUS (2x introns) reporter gene 
            driven by OsUbi promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10053)
  AUTHORS   Hayta S, Smedley MA, Clarke M, Forner M, Harwood WA
  TITLE     An Efficient Agrobacterium-Mediated Transformation Protocol for 
            Hexaploid and Tetraploid Wheat.
  JOURNAL   Curr Protoc. 2021 Mar;1(3):e58. doi: 10.1002/cpz1.58.
  PUBMED    33656289
REFERENCE   2  (bases 1 to 10053)
  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: "Curr 
            Protoc."; date: "2021-03"; volume: "1(3):e58. doi"; pages: " 
            10.1002/cpz1.58"
COMMENT     Sequence Label: pGGG Wheat GUS marker
FEATURES             Location/Qualifiers
     source          1..10053
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..843
                     /label=Act1 promoter
                     /note="rice actin promoter"
     CDS             846..863
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     intron          1745..1934
                     /label=cat1 intron
                     /note="castor bean catalase intron, modified"
     terminator      2634..2886
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     misc_feature    2986..3010
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     misc_feature    3094..3116
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA 
                     (truncated)"
     rep_origin      3243..3678
                     /label=pSa ori
                     /note="origin of replication from bacterial plasmid pSa"
     primer_bind     complement(3958..3977)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             3969..4784
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     CDS             3969..4784
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     complement(4042..4061)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      4955..5543
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5444..5463
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    5634..5658
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     terminator      complement(5720..5972)
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
ORIGIN
        1 tcgaggtcat tcatatgctt gagaagagag tcgggatagt ccaaaataaa acaaaggtaa
       61 gattacctgg tcaaaagtga aaacatcagt taaaaggtgg tataaagtaa aatatcggta
      121 ataaaaggtg gcccaaagtg aaatttactc ttttctacta ttataaaaat tgaggatgtt
      181 tttgtcggta ctttgatacg tcatttttgt atgaattggt ttttaagttt attcgctttt
      241 ggaaatgcat atctgtattt gagtcgggtt ttaagttcgt ttgcttttgt aaatacagag
      301 ggatttgtat aagaaatatc tttaaaaaaa cccatatgct aatttgacat aatttttgag
      361 aaaaatatat attcaggcga attctcacaa tgaacaataa taagattaaa atagctttcc
      421 cccgttgcag cgcatgggta ttttttctag taaaaataaa agataaactt agactcaaaa
      481 catttacaaa aacaacccct aaagttccta aagcccaaag tgctatccac gatccatagc
      541 aagcccagcc caacccaacc caacccaacc caccccagtc cagccaactg gacaatagtc
      601 tccacacccc cccactatca ccgtgagttg tccgcacgca ccgcacgtct cgcagccaaa
      661 aaaaaaaaaa gaaagaaaaa aaagaaaaag aaaaaacagc aggtgggtcc gggtcgtggg
      721 ggccggaaac gcgaggagga tcgcgagcca gcgacgaggc cggccctccc tccgcttcca
      781 aagaaacgcc ccccatcgcc actatataca tacccccccc tctcctccca tccccccaac
      841 cctaccacca ccaccaccac cacctccacc tcctcccccc tcgctgccgg acgacgagct
      901 cctcccccct ccccctccgc cgccgccgcg ccggtaacca ccccgcccct ctcctctttc
      961 tttctccgtt ttttttttcc gtctcggaca cgatctttgg ccttggtagt ttgggtgggc
     1021 gagaggcggc ttcgtgcgcg cccagatcgg tgcgcgggag gggcgggatc tcgcggctgg
     1081 ggctctcgcc ggcgtggatc cggcccggat ctcgcgggga atggggctct cggatgtaga
     1141 tctgcgatcc gccgttgttg ggggagatga tggggggttt aaaatttccg ccatgctaaa
     1201 caagatcagg aagaggggaa aagggcacta tggtttatat ttttatatat ttctgctgct
     1261 tcgtcaggct tagatgtgct agatctttct ttcttctttt tgtgggtaga atttgaatcc
     1321 ctcagcattg ttcatcggta gtttttcttt tcatgatttg tgacaaatgc agcctcgtgc
     1381 ggagcttttt tgtaggtaga agccaatgaa aaagcctgaa ctcaccgcga cgtctgtcga
     1441 gaagtttctg atcgaaaagt tcgacagcgt ctccgacctg atgcagctct cggagggcga
     1501 agaatctcgt gctttcagct tcgatgtagg agggcgtgga tatgtcctgc gggtaaatag
     1561 ctgcgccgat ggtttctaca aagatcgtta tgtttatcgg cactttgcat cggccgcgct
     1621 cccgattccg gaagtgcttg acattgggga attcagcgag agcctgacct attgcatctc
     1681 ccgccgtgca cagggtgtca cgttgcaaga cctgcctgaa accgaactgc ccgctgttct
     1741 gcaggtaaat ttctagtttt tctccttcat tttcttggtt aggacccttt tctcttttta
     1801 tttttttgag ctttgatctt tctttaaact gatctatttt ttaattgatt ggttatggtg
     1861 taaatattac atagctttaa ctgataatct gattacttta tttcgtgtgt ctatgatgat
     1921 gatgataact gcagccggtc gcggaggcca tggatgcgat cgctgcggcc gatcttagcc
     1981 agacgagcgg gttcggccca ttcggaccgc aaggaatcgg tcaatacact acatggcgtg
     2041 atttcatatg cgcgattgct gatccccatg tgtatcactg gcaaactgtg atggacgaca
     2101 ccgtcagtgc gtccgtcgcg caggctctcg atgagctgat gctttgggcc gaggactgcc
     2161 ccgaagtccg gcacctcgtg cacgcggatt tcggctccaa caatgtcctg acggacaatg
     2221 gccgcataac agcggtcatt gactggagcg aggcgatgtt cggggattcc caatacgagg
     2281 tcgccaacat cttcttctgg aggccgtggt tggcttgtat ggagcagcag acgcgctact
     2341 tcgagcggag gcatccggag cttgcaggat cgccgcggct ccgggcgtat atgctccgca
     2401 ttggtcttga ccaactctat cagagcttgg ttgacggcaa tttcgatgat gcagcttggg
     2461 cgcagggtcg atgcgacgca atcgtccgat ccggagccgg gactgtcggg cgtacacaaa
     2521 tcgcccgcag aagcgcggcc gtctggaccg atggctgtgt agaagtactc gccgatagtg
     2581 gaaaccgacg ccccagcact cgtccgaggg caaaggaata ggcttgtcaa gcagatcgtt
     2641 caaacatttg gcaataaagt ttcttaagat tgaatcctgt tgccggtctt gcgatgatta
     2701 tcatataatt tctgttgaat tacgttaagc atgtaataat taacatgtaa tgcatgacgt
     2761 tatttatgag atgggttttt atgattagag tcccgcaatt atacatttaa tacgcgatag
     2821 aaaacaaaat atagcgcgca aactaggata aattatcgcg cgcggtgtca tctatgttac
     2881 tagatcgacg ctggatccga attcggcatg agacccacag agtgttcaac cccagtacat
     2941 taaaaacgtc cgcaatgtgt tattaagttg tctaagcgtc aatttgttta caccacaata
     3001 tatcctgcca ccagccagcc aacagctccc cgaccggcag ctcggcacaa aatcaccact
     3061 cgatacaggc agcccatcaa ccatgactaa acgttacacc acaatatatc ctgccaagat
     3121 ctaattccgg ggatcggaaa tccagaagcc cgagaggttg ccgcctttcg ggctttttct
     3181 ttttcaaaaa aaaaaattta taaaacgatc tgttgcggcc ggccgccggg ttgtgggcaa
     3241 aggcgctcga cggtgggcaa ccgcttgcgg ttgtccacgg gcggagccgg tgcgcgtagc
     3301 gcattgtcca caagccaagg gcgaccaata attgatatat atattcataa ttgaaaagct
     3361 aattgaacat actacttgct gtaactactt gccggagcga ggggtgtttg caagctgttg
     3421 atctgaaagg gctattagcg ttctcacgtg cctttttgat tagcgatttc acgtgacctt
     3481 attagcgatt tcacgtactc cgattagcga tttcacgtac cctgattagc gatttcacgt
     3541 ggatagtttt tggagcgggc cggaaagccc cgtgaatcaa ggctttgcgg ggcattagcg
     3601 gtttcacgtg gataactacc ctctatccac aggcttccgg ggataaaaaa gcccgctcga
     3661 cggcgggctg ttggatgggg atcgcctgaa tcgccccatc atccagccag aaagtgaggg
     3721 agccacggtt gatgagagct ttgttgtagg tggaccagtt ggggattttg aacttttgct
     3781 ttgccacgga acggtctgcg ttgtcgggaa gatgcgtgat ctgatccttc aactcagcaa
     3841 aagttcgatt tattcaacaa agccacgttg tgtctcaaaa tctctgatgt tacattgcac
     3901 aagataaaaa tatatcatca tgaacaataa aactgtctgc ttacataaac agtaatacaa
     3961 ggggtgttat gagccatatt caacgggaaa cgtcttgctc aaggccgcga ttaaattcca
     4021 acatggatgc tgatttatat gggtataaat gggctcgcga taatgtcggg caatcaggtg
     4081 cgacaatcta ccgattgtat gggaagcccg atgcgccaga gttgtttctg aaacatggca
     4141 aaggtagcgt tgccaatgat gttacagatg agatggtcag actaaactgg ctgacggaat
     4201 ttatgcctct tccgaccatc aagcatttta tccgtactcc tgatgatgca tggttactca
     4261 ccactgcgat cccagggaaa acagcattcc aggtattaga agaatatcct gattcaggtg
     4321 aaaatattgt tgatgcgctg gcagtgttcc tgcgccggtt gcattcgatt cctgtttgta
     4381 attgtccttt taacagcgat cgcgtatttc gtctcgctca ggcgcaatca cgaatgaata
     4441 acggtttggt tgatgcgagt gattttgatg acgagcgtaa tggctggcct gttgaacaag
     4501 tctggaaaga aatgcataaa cttttgccat tctcaccgga ttcagtcgtc actcatggtg
     4561 atttctcact tgataacctt atttttgacg aggggaaatt aataggttgt attgatgttg
     4621 gacgagtcgg aatcgcagac cgataccagg atcttgccat cctatggaac tgcctcggtg
     4681 agttttctcc ttcattacag aaacggcttt ttcaaaaata tggtattgat aatcctgata
     4741 tgaataaatt gcagtttcat ttgatgctcg atgagttttt ctaatcacta gaccaatgtt
     4801 acacatatat actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga
     4861 agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag
     4921 cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa
     4981 tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag
     5041 agctaccaac tcttcttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg
     5101 ttcttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat
     5161 acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta
     5221 ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg
     5281 gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc
     5341 gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa
     5401 gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc
     5461 tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt
     5521 caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct
     5581 tttgctggcc ttttgctcac atgagatctc aaacaaacac atacagcgac ttagtttacc
     5641 cgccaatata tcctgtcaag gaaacggatc acttcgtgtc cctcggtcac atgtgcatcc
     5701 tcggtctcag taaagcgtcg atctagtaac atagatgaca ccgcgcgcga taatttatcc
     5761 tagtttgcgc gctatatttt gttttctatc gcgtattaaa tgtataattg cgggactcta
     5821 atcataaaaa cccatctcat aaataacgtc atgcattaca tgttaattat tacatgctta
     5881 acgtaattca acagaaatta tatgataatc atcgcaagac cggcaacagg attcaatctt
     5941 aagaaacttt attgccaaat gtttgaacga tctgcttgac aagctcattg tttgcctccc
     6001 tgctgcggtt tttcaccgaa gttcatgcca gtccagcgtt tttgcagcag aaaagccgcc
     6061 gacttcggtt tgcggtcgcg agtgaagatc cctttcttgt taccgccaac gcgcaatatg
     6121 ccttgcgagg tcgcaaaatc ggcgaaattc catacctgtt caccgacgac ggcgctgacg
     6181 cgatcaaaga cgcggtgata catatccagc catgcacact gatactcttc actccacatg
     6241 tcggtgtaca ttgagtgcag cccggctaac gtatccacgc cgtattcggt gatgataatc
     6301 ggctgatgca gtttctcctg ccaggccaga agttcttttt ccagtacctt ctctgccgtt
     6361 tccaaatcgc cgctttggac ataccatccg taataacggt tcaggcacag cacatcaaag
     6421 agatcgctga tggtatcggt gtgagcgtcg cagaacatta cattgacgca ggtgatcgga
     6481 cgcgtcgggt cgagtttacg cgttgcttcc gccagtggcg cgaaatattc ccgtgcacct
     6541 tgcggacggg tatccggttc gttggcaata ctccacatca ccacgcttgg gtggtttttg
     6601 tcacgcgcta tcagctcttt aatcgcctgt aagtgcgctt gctgagtttc cccgttgact
     6661 gcctcttcgc tgtacagttc tttcggcttg ttgcccgctt cgaaaccaat gcctaaagag
     6721 aggttaaagc cgacagcagc agtttcatca atcaccacga tgccatgttc atctgcccag
     6781 tcgagcatct cttcagcgta agggtaatgc gaggtacggt aggagttggc cccaatccag
     6841 tccattaatg cgtggtcgtg caccatcagc acgttatcga atcctttgcc acgcaagtcc
     6901 gcatcttcat gacgaccaaa gccagtaaag tagaacggtt tgtggttaat caggaactgt
     6961 tcgcccttca ctgccactga ccggatgccg acgcgaagcg ggtagatatc acactctgtc
     7021 tggcttttgg ctgtgacgca cagttcatag agataacctt cacccggttg ccagaggtgc
     7081 ggattcacca cttgcaaagt cccgctagtg ccttgtccag ttgcaaccac ctgttgatcc
     7141 gcatcacgca gttcaacgct gacatcacca ttggccacca cctgtaaggg gaaaaaaatt
     7201 gacaaaaagc aaaggtcaag caaatgtatg gaaatataaa atttcacaca gtatctctta
     7261 agttacacgt tgaagcatga agtacctgcc agtcaacaga cgcgtggtta cagtcttgcg
     7321 cgacatgcgt caccacggtg atatcgtcca cccaggtgtt cggcgtggtg tagagcatta
     7381 cgctgcgatg gattccggca tagttaaaga aatcatggaa gtaagactgc tttttcttgc
     7441 cgttttcgtc ggtaatcacc attcccggcg ggatagtctg ccagttcagt tcgttgttca
     7501 cacaaacggt gatacgtaca cttttcccgg caataacata cggcgtgaca tcggcttcaa
     7561 atggcgtata gccgccctga tgctccatca cttcctgatt attgacccac actttgccgt
     7621 aatgagtgac cgcatcgaaa cgcagcacga tacgctggcc tgcccaacct gaaaaaagat
     7681 tattagcgtc agtacaaata gtatacttca aaaaattaaa aatacagacc aaaaaagaga
     7741 gagtaagact tacctttcgg tataaagact tcgcgctgat accagacgtt gcccgcataa
     7801 ttacgaatat ctgcatcggc gaactgatcg ttaaaactgc ctggcacagc aattgcccgg
     7861 ctttcttgta acgcgctttc ccaccaacgc tgatcaattc cacagttttc gcgatccaga
     7921 ctgaatgccc acaggccgtc gagttttttg atttcacggg ttggggtttc tacaggacgt
     7981 aacataaggg actgacccat ttatctggat ctggaagagg caagaaagga ttggaattaa
     8041 cataggtaaa aaaaagcagc tggagaataa gttggcacca gatcctacga tatcggtact
     8101 gtcagctcaa aatagatgaa catatcaatc acatcatact acactacaat tggacagcat
     8161 cattaatcat aagagcgctg ctcacagctc acaccacatc aacaagcaac aaccaaaaat
     8221 atcttcagat aataacaaaa ctgacataag taactacagg acaacagatc tatgtaagaa
     8281 cacagaataa agccacctag ttctagcatc tgacagattt gatttgcacg agtaaagcaa
     8341 ataaaacggg taaaaaaaca cagtacaaca tcagcactac agttcaacta ggcaaagagc
     8401 cagatacaat acagattaaa ttgaccttag aacccaagct ctacagttca agtaggtgta
     8461 cacgatacag atgtaattct ctaacccagc tccacacaat tccaatccaa atccagggga
     8521 aaaaaactaa gaacacaatt ccaatccaaa tccagaggaa ataaactatg tgtcgcctaa
     8581 ctcaacctaa agattttaac ctaacgagct attcatctat tggaacaata tcagaacaat
     8641 cccatccgaa ctaagcacaa ataagtaaaa ctaaatctga tcacaaaact caaactatgt
     8701 ggcaggatca aaacataact gtgaactact cttattatcc atcattaggg taatatatcg
     8761 aaacaatccc catctgacac atcattaggg taatatatcg aaacaatccc catctgacac
     8821 atcattaggg taatatatcg aaacaatccc catctgacag aacaatctca aaaccaatcc
     8881 atctattaga tcgatataaa taacacagat ccctacctcg attctcacga aaccctaaat
     8941 caatgagaga ttaagaggat taccctaaaa tcgcacgaaa ccgtaactcg attaagaggt
     9001 ggatcaccct aaaatcgcac gaaaccctga gagattgaga ctaactcgca cgggatcctc
     9061 ctacgctttc caccacgagc atcaaaatcg cgagattaga cgggcaccac gagagattcg
     9121 aggattagga cgaggtgatt gattagttac cttgacgagg cgattagaga acgcaaagct
     9181 tcgctccccc tccccttcgg tcgattcgac tcgctatgcg attaatggag agaaaacctc
     9241 tctatataaa gcgtgggcaa ggcggtgcga ggcagcatcc ggtgcggtca ccgccacgcg
     9301 tacggcccaa tggcagcacc acacgtgtcc atgtagttgg aggcgcgaca cgtgtccggt
     9361 cccggtacga ccatccagac ggtgcccgga accaactccg cgtcaggtca tgccgcctct
     9421 ctttgacgaa gtcggcatgc ctgatcaaga ccgtacgttt ccgatccaac gatcaggaaa
     9481 catcgtactt gatctgctat aagctgcgtg tgggccttaa taatcctctg gggaaatatt
     9541 tgtgggctga aatgaacggg atattgggcc tagacaaacc agggcccatg aattttacgg
     9601 cccaatagat tgttgacacc taattggcta agcccaactc gcgcactctg accaaccgat
     9661 cggaagttcg cgtccacgcc gccctgctcc tccgtcgccg ccggctcggc ggctgacccc
     9721 cgcgccgccg caccagctgc gcgcgcgcgc gcgcgcgccc ccaccaccgc ggcctctccc
     9781 tcccacgcgc agagggcacg ggcagaggcc gccgcggcgc cgtactcgcc ggcgcttcgt
     9841 cggcctcggc ctcgggtcga ggccggtggc ggggcctcgt cagatcgccc cccttcctgt
     9901 gcagtaagta tttcccacgg acccaatatg gattcgttgt tctaatcgat agctgctatc
     9961 agagatgggt tcttctgcgt gatttacctg ttcttcacat cctccgagct cgaattctag
    10021 taaacccgtc aaaaccttgc ggagctagca tac
//
