LOCUS       sequence-317401-        3774 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Vitis miR396-resistant GRF4-GIF1 in pDONR-zeo.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3774)
  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 3774)
  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: JD638
FEATURES             Location/Qualifiers
     source          1..3774
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     27..44
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(203..230)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(322..408)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     457..479
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     471..488
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     472..488
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    504..603
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     protein_bind    complement(2305..2404)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2421..2440)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2422..2440)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2445..2461)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2445..2461)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             complement(2529..2903)
                     /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(2922..2969)
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     rep_origin      3059..3647
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3548..3567
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg gaagagcgcc
       61 caatacgcaa accgcctctc cccgcgcgtt ggccgattca ttaatgcagc tggcacgaca
      121 ggtttcccga ctggaaagcg ggcagtgagc gcaacgcaat taatacgcgt accgctagcc
      181 aggaagagtt tgtagaaacg caaaaaggcc atccgtcagg atggccttct gcttagtttg
      241 atgcctggca gtttatggcg ggcgtcctgc ccgccaccct ccgggccgtt gcttcacaac
      301 gttcaaatcc gctcccggcg gatttgtcct actcaggaga gcgttcaccg acaaacaaca
      361 gataaaacga aaggcccagt cttccgactg agcctttcgt tttatttgat gcctggcagt
      421 tccctactct cgcgttaacg ctagcatgga tgttttccca gtcacgacgt tgtaaaacga
      481 cggccagtct taagctcggg ccccaaataa tgattttatt ttgactgata gtgacctgtt
      541 cgttgcaaca aattgatgag caatgctttt ttataatgcc aactttgtac aaaaaagctg
      601 ccaccatgaa gcaaagcttt gtgggggttt cttcacaaac agaggctgag acaagagcca
      661 gagagagtga cacacagaca cacccacaaa ggaacctcag tcactgtccc caaaaacatc
      721 ctcaaaactt gcaaagaacc caaaatggtg ggtctgttgg gggggcatca atggcaacag
      781 ggatgatggt gaggtcatca ccattcacag tgtctcagtg gcaggagctg gagcatcaag
      841 ctttgatctt taagtatcta atggcaggac tacccgtccc acctgatctt gtgcttccta
      901 ttcagaagag ctttgaatct atgtcttcta ggttcttcca ccatcctacc atgggttact
      961 gttccttcta tgggaagaag gtggacccgg agccaggtag atgccggagg actgatggca
     1021 agaaatggag gtgctccaaa gacgcctacc cggactccaa gtactgtgag cgccacatgc
     1081 accgtggccg caaccgttct agaaaaccgg tcgaatcaca aactactaca cagtcttctt
     1141 cgactgtgac ttcactaact gtcactggag gcagtggcag tggagccggg accttccaga
     1201 accttcccct gcaaaacttc aataaccccc aaggcggccc ggggtccgga accaaccaat
     1261 cccaattcca tatggagccc attccctatg gcattcctaa taaagattac aggtatcttc
     1321 atggacttaa acccgaggta ggcgaacata gtttcttctc tgaagcttca ggaagcaaca
     1381 ggggtcttca gatggactcg cctttggaca gcacatggcc tctgatgcca tcacgggtct
     1441 catcattccc cccatcaaaa tcaagtgaaa actccatctt gcagagtgac tacccccagc
     1501 attcgttctt cagcagcgaa tttgcgtcgg gagaggttgt gaaacaggag ggtcagtctc
     1561 tccgtccttt ctttgacgag tggcctaaaa ccagagactc ctggtccggc cttgaggaag
     1621 agaggtccaa ccagacctca ttctccacaa cccagctctc aatttcgatc ccaatggcct
     1681 cctctgactt ctcaacaacc agctctcggt ctccccacga taacgcggcc gctgccatgc
     1741 agcccatgat ggcagcctat taccccagca acgtcaccac tgatcacatt cagcagtatc
     1801 ttgatgaaaa caagtcattg attctgaaga ttgttgagag ccagaattca ggaaaattga
     1861 ctgaatgtgc agagaaccag gcaagactac agagaaacct catgtacctg gctgcaatcg
     1921 ctgattctca accccaacca cccaccatgc atgctcagtt ccctcctagt ggcattgttc
     1981 agccaggagc tcactacatg cagcaccaac aagctcaaca aatgacacca cagtcgctcc
     2041 tggctgcacg ctcctccatg ctgtactccc aacaaccatt ttcggccctg caacaacaac
     2101 aagccatcca tagccagctt ggcatgggct ctggtggaag tgcaggactt cacatgctgc
     2161 aaagcgaggg gagtaatcca ggaggcaatg gaacactggg gactgatatg ggaaatgcag
     2221 aagggcgagg agggaactca ggaggtcagg gtggggatgg aggtgagact ctttacttga
     2281 aagctgctga agatgggaat tgacgttcag ctttcttgta caaagttggc attataagaa
     2341 agcattgctt atcaatttgt tgcaacgaac aggtcactat cagtcaaaat aaaatcatta
     2401 tttgccatcc agctgatatc ccctatagtg agtcgtatta catggtcata gctgtttcct
     2461 ggcagctctg gcccgtgtct caaaatctct gatgttacat tgcacaagat aaaaatatat
     2521 catcatgatc agtcctgctc ctcggccacg aagtgcacgc agttgccggc cgggtcgcgc
     2581 agggcgaact cccgccccca cggctgctcg ccgatctcgg tcatggccgg cccggaggcg
     2641 tcccggaagt tcgtggacac gacctccgac cactcggcgt acagctcgtc caggccgcgc
     2701 acccacaccc aggccagggt gttgtccggc accacctggt cctggaccgc gctgatgaac
     2761 agggtcacgt cgtcccggac cacaccggcg aagtcgtcct ccacgaagtc ccgggagaac
     2821 ccgagccggt cggtccagaa ctcgaccgct ccggcgacgt cgcgcgcggt gagcaccgga
     2881 acggcactgg tcaacttggc catggtttag ttcctcacct tgtcgtatta tactatgccg
     2941 atatactatg ccgatgatta attgtcaaca cgtgctgatc atgaccaaaa tcccttaacg
     3001 tgagttacgc gtcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt
     3061 gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag
     3121 cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta actggcttca
     3181 gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca
     3241 agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg
     3301 ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg
     3361 cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct
     3421 acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga
     3481 gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc
     3541 ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg
     3601 agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg
     3661 cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc tttcctgcgt
     3721 tatcccctga ttctgtggat aaccgtatta ccgcctttga gtgagctgat accg
//
