LOCUS       sequence-417750-        3092 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mutant E3-DART degron casette for N-terminal fusion to protein of 
            interest using Goldenbraid.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3092)
  AUTHORS   Huang L, Rojas-Pierce M
  TITLE     Rapid depletion of target proteins in plants by an inducible protein
            degradation system.
  JOURNAL   Plant Cell. 2024 Mar 6:koae072. doi: 10.1093/plcell/koae072.
  PUBMED    38446628
REFERENCE   2  (bases 1 to 3092)
  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.
            2024 Mar 6:koae072. doi: 10.1093/plcell/koae072."
COMMENT     Sequence Label: pUPD2-HR1bR181A R185A-GFP-L (B3-B4)
FEATURES             Location/Qualifiers
     source          1..3092
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             join(265..267,268..270,271..981)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(265..267,268..270,271..981)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(310..331)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(571..590)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     918..939
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     terminator      1049..1106
                     /label=his operon terminator
                     /note="This putative transcriptin terminator from the E. 
                     coli his operon has a 2-bp deletion introduced during 
                     synthesis. Its efficiency has not been determined."
     primer_bind     complement(1130..1147)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1301..1889)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1381..1400)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(2072..2166)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(2187..2846)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     2761..2780
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     primer_bind     3008..3026
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     terminator      complement(3030..3073)
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
     misc_feature    join(3076..3092,1..5)
                     /label=BioBrick prefix
                     /note="BioBrick prefix for parts that do not start with 
                     ""ATG"""
ORIGIN
        1 tagagcgatg agtggtctcg aatggctact aatctttcta gagttgctgg acttgaaaag
       61 caacttgcta ttgaacttaa ggttaagcaa ggtgctgaaa atatgattca aacttattct
      121 aatggatcta ctaaggatag aaagcttctt cttactgctc aacaaatgct tcaagattct
      181 aagactaaga ttgatattat tgcaatgcaa cttgcaagag ctcttcaagc tggtcaactt
      241 gaaaatcaag ctgctcctgg ttcgatggtg agcaagggcg aggagctgtt caccggggtg
      301 gtgcccatcc tggtcgagct ggacggcgac gtaaacggcc acaagttcag cgtgtccggc
      361 gagggcgagg gcgatgccac ctacggcaag ctgaccctga agttcatctg caccaccggc
      421 aagctgcccg tgccctggcc caccctcgtg accaccctga cctacggcgt gcagtgcttc
      481 agccgctacc ccgaccacat gaagcagcac gacttcttca agtccgccat gcccgaaggc
      541 tacgtccagg agcgcaccat cttcttcaag gacgacggca actacaagac ccgcgccgag
      601 gtgaagttcg agggcgacac cctggtgaac cgcatcgagc tgaagggcat cgacttcaag
      661 gaggacggca acatcctggg gcacaagctg gagtacaact acaacagcca caacgtctat
      721 atcatggccg acaagcagaa gaacggcatc aaggtgaact tcaagatccg ccacaacatc
      781 gaggacggca gcgtgcagct cgccgaccac taccagcaga acacccccat cggcgacggc
      841 cccgtgctgc tgcccgacaa ccactacctg agcacccagt ccgccctgag caaagacccc
      901 aacgagaagc gcgatcacat ggtcctgctg gagttcgtga ccgccgccgg gatcactctc
      961 ggcatggacg agctgtacaa gggaggtggc tcatctggcg gaggtggttc gtgagaccac
     1021 tcatcgcgac tagtagcggc cgctgcagtc cggcaaaaaa gggcaaggtg tcaccaccct
     1081 gccctttttc tttaaaaccg aaaagattac ttcgcgttat gcaggcttcc tcgctcactg
     1141 actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa
     1201 tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc
     1261 aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccacagg ctccgccccc
     1321 ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat
     1381 aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc
     1441 cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct
     1501 cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg
     1561 aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc
     1621 cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga
     1681 ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa
     1741 gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta
     1801 gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc
     1861 agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg
     1921 acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga
     1981 tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg
     2041 agtaaacttg gtctgacagc tcgaggcttg gattctcacc aataaaaaac gcccggcggc
     2101 aaccgagcgt tctgaacaaa tccagatgga gttctgaggt cattactgga tctatcaaca
     2161 ggagtccaag cgagctcgat atcaaattac gccccgccct gccactcgtc gcagtactgt
     2221 tgtaattcat taagcattct gccgacatgg aagccatcac aaacggcatg atgaacctga
     2281 atcgccagcg gcatcagcac cttgtcgcct tgcgtataat atttgcccat ggtgaaaacg
     2341 ggggcgaaga agttgtccat attggccacg tttaaatcaa aactggtgaa actcacccag
     2401 ggattggctg atacgaaaaa catattctca ataaaccctt tagggaaata ggccaggttt
     2461 tcaccgtaac acgccacatc ttgcgaatat atgtgtagaa actgccggaa atcgtcgtgg
     2521 tattcactcc agagtgatga aaacgtttca gtttgctcat ggaaaacggt gtaacaaggg
     2581 tgaacactat cccatatcac cagctcaccg tctttcattg ccatacgaaa ttccggatga
     2641 gcattcatca ggcgggcaag aatgtgaata aaggccggat aaaacttgtg cttatttttc
     2701 tttacggtct ttaaaaaggc cgtaatatcc agctgaacgg tctggttata ggtacattga
     2761 gcaactgact gaaatgcctc aaaatgttct ttacgatgcc attgggatat atcaacggtg
     2821 gtatatccag tgattttttt ctccatttta gcttccttag ctcctgaaaa tctcgataac
     2881 tcaaaaaata cgcccggtag tgatcttatt tcattatggt gaaagttgga acctcttacg
     2941 tgcccgatca actcgagtgc cacctgacgt ctaagaaacc attattatca tgacattaac
     3001 ctataaaaat aggcgtatca cgaggcagaa tttcagataa aaaaaatcct tagctttcgc
     3061 taaggatgat ttctggaatt cgcggccgct tc
//
