LOCUS       sequence-317235-        3987 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Wheat GRF4-GIF1 chimera in pDONR-zeo.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3987)
  AUTHORS   Debernardi JM, Tricoli DM, Ercoli MF, Hayta S, Ronald P, Palatnik 
            JF, Dubcovsky J
  TITLE     A GRF-GIF chimeric protein improves the regeneration efficiency of 
            transgenic plants.
  JOURNAL   Nat Biotechnol. 2020 Oct 12. pii: 10.1038/s41587-020-0703-0. doi: 
            10.1038/s41587-020-0703-0.
  PUBMED    33046875
REFERENCE   2  (bases 1 to 3987)
  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: "Nat 
            Biotechnol. 2020 Oct 12. pii: 10.1038/s41587-020-0703-0. doi: 
            10.1038/s41587-020-0703-0."
COMMENT     Sequence Label: JD553
FEATURES             Location/Qualifiers
     source          1..3987
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     95..112
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(271..298)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(390..476)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     525..547
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     539..556
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     540..556
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    572..671
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     regulatory      668..677
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    complement(2586..2685)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2702..2721)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2703..2721)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2726..2742)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2726..2742)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             complement(2810..3184)
                     /codon_start=1
                     /gene="Sh ble from Streptoalloteichus hindustanus"
                     /product="antibiotic-binding protein"
                     /label=BleoR
                     /note="confers resistance to bleomycin, phleomycin, and 
                     Zeocin(TM)"
                     /translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD
                     VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR
                     EFALRDPAGNCVHFVAEEQD"
     promoter        complement(3203..3250)
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     rep_origin      3340..3928
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3829..3848
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gttctttcct gcgttatccc ctgattctgt ggataaccgt attaccgcct ttgagtgagc
       61 tgataccgct cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga
      121 agagcgccca atacgcaaac cgcctctccc cgcgcgttgg ccgattcatt aatgcagctg
      181 gcacgacagg tttcccgact ggaaagcggg cagtgagcgc aacgcaatta atacgcgtac
      241 cgctagccag gaagagtttg tagaaacgca aaaaggccat ccgtcaggat ggccttctgc
      301 ttagtttgat gcctggcagt ttatggcggg cgtcctgccc gccaccctcc gggccgttgc
      361 ttcacaacgt tcaaatccgc tcccggcgga tttgtcctac tcaggagagc gttcaccgac
      421 aaacaacaga taaaacgaaa ggcccagtct tccgactgag cctttcgttt tatttgatgc
      481 ctggcagttc cctactctcg cgttaacgct agcatggatg ttttcccagt cacgacgttg
      541 taaaacgacg gccagtctta agctcgggcc ccaaataatg attttatttt gactgatagt
      601 gacctgttcg ttgcaacaaa ttgatgagca atgctttttt ataatgccaa ctttgtacaa
      661 aaaagctgcc accatggcga tgccgtatgc ctctctttcc ccggcaggcg accgccgctc
      721 ctccccggcc gccaccgcca ccgcctccct cctccccttc tgccgctcct cccccttctc
      781 cgccggcggc aatggcggca tgggggagga ggcgcggatg gacgggaggt ggatggcgag
      841 gccggtgccc ttcacggcgg cgcagtacga ggagctggag caccaggcgc tcatatacaa
      901 gtacctggtg gccggcgtgt ccgtcccgcc ggatctcgtg ctccccatcc gccgcggcat
      961 cgagtccctc gccgcccgct tctaccacaa ccccctcgcc atcgggtacg gatcgtacct
     1021 gggcaagaag gtggatccgg agccgggccg gtgccggcgc acggacggca agaagtggcg
     1081 gtgcgccaag gaggccgcct ccgactccaa gtattgcgag cgccacatgc accgcggccg
     1141 caaccgttca agaaagcctg tggaaacgca gctcgtctcg cactcccagc cgccggccgc
     1201 ctccgtcgtg ccgcccctcg ccaccggctt ccacaaccac tccctctacc ccgccatcgg
     1261 cggcaccaac ggtggtggag gcggggggaa caacggcatg tccatgcccg gcacgttctc
     1321 ctccgcgctg gggccgcctc agcagcacat gggcaacaat gccgcctctc cctacgcggc
     1381 tctcggcggc gccggaacat gcaaagattt caggtatacc gcatatggaa taagatcttt
     1441 ggcagacgag cagagtcagc tcatgacaga agccatgaac acctccgtgg agaacccatg
     1501 gcgcctgccg ccatcttctc aaacgactac attcccgctc tcaagctact ctcctcagct
     1561 tggagcaacg agtgacctgg gtcagaacaa cagcagcaac aacaacagcg gcgtcaaggc
     1621 cgagggacag cagcagcagc agccgctctc cttcccgggg tgcggcgact tcggcagcgg
     1681 cgactccgcg aagcaggaga accagacgct gcggccgttc ttcgacgagt ggccgaagac
     1741 gagggactcg tggtcggacc tgaccgacga caactcgaac gtcgcctcct tctcggccac
     1801 ccagctgtcg atctcgatac ccatgacgtc ctccgacttc tccgccgcca gctcccagtc
     1861 gcccaacggc atgctgttcg ccggcgaaat gtacgcggcc gctgccatgc agcagcaaca
     1921 cctgatgcag atgaaccaga gcatgatggg gggctacgct tcctctacca ctgccaccac
     1981 tgatctcatt cagcagtacc tggatgagaa caagcagctg atcctggcca tcctcgacaa
     2041 ccagaacaac ggcaaggtgg aggagtgcgc acggaaccaa gctaagctcc agcagaacct
     2101 catgtacctc gccgccatcg ccgacagcca gcctccgcag acggcatcgc tgtctcagta
     2161 cccgtccaac ctgatgatgc agtccgggcc gcggtacatg cagcagcagt cggcgcagat
     2221 gatgtcgccg cagtcgctga tggcggcgcg gtcgtcgatg atgtacgcgc agcaggccat
     2281 gtcgccgctc cagcagcagc agcagcagca gcagcaccag gcggccgcgc acggccagct
     2341 ggggatgtcc tccggcgcga ccaccgggtt caacctcctg cacggtgagg ccagcatggg
     2401 cggcggcggc ggcgccactg gcaacagcat gatgaacgcc agcgtcttct cggactatgg
     2461 ccgcggcggc agcggcgcca aggaggggtc gacctcgctg tcggccgacg ctcgcggcgc
     2521 caactctggc gcgcacagcg gcgacgggga gtacctcaag ggcaccgagg aggaaggaag
     2581 ctagcgttca gctttcttgt acaaagttgg cattataaga aagcattgct tatcaatttg
     2641 ttgcaacgaa caggtcacta tcagtcaaaa taaaatcatt atttgccatc cagctgatat
     2701 cccctatagt gagtcgtatt acatggtcat agctgtttcc tggcagctct ggcccgtgtc
     2761 tcaaaatctc tgatgttaca ttgcacaaga taaaaatata tcatcatgat cagtcctgct
     2821 cctcggccac gaagtgcacg cagttgccgg ccgggtcgcg cagggcgaac tcccgccccc
     2881 acggctgctc gccgatctcg gtcatggccg gcccggaggc gtcccggaag ttcgtggaca
     2941 cgacctccga ccactcggcg tacagctcgt ccaggccgcg cacccacacc caggccaggg
     3001 tgttgtccgg caccacctgg tcctggaccg cgctgatgaa cagggtcacg tcgtcccgga
     3061 ccacaccggc gaagtcgtcc tccacgaagt cccgggagaa cccgagccgg tcggtccaga
     3121 actcgaccgc tccggcgacg tcgcgcgcgg tgagcaccgg aacggcactg gtcaacttgg
     3181 ccatggttta gttcctcacc ttgtcgtatt atactatgcc gatatactat gccgatgatt
     3241 aattgtcaac acgtgctgat catgaccaaa atcccttaac gtgagttacg cgtcgttcca
     3301 ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg
     3361 cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga
     3421 tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa
     3481 tactgttctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc
     3541 tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg
     3601 tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac
     3661 ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct
     3721 acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc
     3781 ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg
     3841 gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg
     3901 ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct
     3961 ggccttttgc tggccttttg ctcacat
//
