LOCUS       sequence-334925-        2846 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Donor plasmid containing the HDR template for the in-frame, 
            N-terminal 2xFLAG-mAID tagging of AMBRA1 gene.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 2846)
  AUTHORS   Simoneschi D, Rona G, Zhou N, Jeong YT, Jiang S, Milletti G, Arbini 
            AA, O'Sullivan A, Wang AA, Nithikasem S, Keegan S, Siu Y, 
            Cianfanelli V, Maiani E, Nazio F, Cecconi F, Boccalatte F, Fenyo D, 
            Jones DR, Busino L, Pagano M
  TITLE     CRL4(AMBRA1) is a master regulator of D-type cyclins.
  JOURNAL   Nature. 2021 Apr;592(7856):789-793. doi: 10.1038/s41586-021-03445-y.
            Epub 2021 Apr 14.
  PUBMED    33854235
REFERENCE   2  (bases 1 to 2846)
  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"; doi: 
            "10.1038/s41586-021-03445-y"; journalName: "Nature"; date: 
            "2021-04"; volume: "592"; issue: "7856"; pages: "789-793"
COMMENT     Sequence Label: pUC57-2xFLAG-mAID-AMBRA1 HDR donor template
FEATURES             Location/Qualifiers
     source          1..2846
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     CDS             1199..1222
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             1238..1261
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             1307..1510
                     /codon_start=1
                     /product="minimal degron from the auxin-responsive IAA17 
                     protein of Arabidopsis thaliana (Kubota et al., 2013)"
                     /label=mini-AID
                     /note="acts as an auxin-inducible degron (AID) in the 
                     presence of TIR1"
                     /translation="KEKSACPKDPAKPPAKAQVVGWPPVRSYRKNVMVSCQKSSGGPEA
                     AAFVKVSMDGAPYLRKIDLRMYK"
ORIGIN
        1 aacatccttc ttcctaggag ttgcataatg aattaaatat gcagggttaa agtgtcttga
       61 tacctactga aataattcca aaattataca aaaaaaacac caacctcaat gtaaccatat
      121 ctctatccat ggagatagat gaggcaaatt tggcaaaata ttgacaattc ttgagtctag
      181 gtggagaata gtgtatgttt attgtaccat tctttcaact ttcctatatt ttttaaaaca
      241 ttcatagtgg agctaaagaa acaatgtcac agtggataac actgtatata tgtagatatt
      301 cttttgaaat accattttat caggcttaca tagtccattg tatagaccac aatttgttct
      361 ttaatcatta cacttgtgtg tattgtgctg ctagttgcca ttttagtaga attttaattt
      421 ctgatgaagt tttctccttg taagtcttgt ggtagatgag aaaaggagcc tgattttttt
      481 ttttccttcc ctttccttct ctgtcctcta ccctggatct ttagtggaga agagaaagag
      541 aaaatgagtg agaaaggtac tcctaaataa tctttttttt tttgagatgg agtcttgctc
      601 tgtcgcccag gcactggagt gcaatggcac tatctcagct ctctgcaact tctgcgtcct
      661 gggttcaagc aattctcctg cttcagtctg atgagtagct gggattacag gtgcctgcca
      721 ccacacccgg ctaatttttg tattttagta gagacggggt ttcaccatgt tggccaggct
      781 gtctcaaact cctgacctca agcaatccaa tctccttggc ttcccaaagt tcggggatta
      841 caggcgtgag ccactgtgct tggcctgatc ctttctttag ctacaaggtt gagctatgga
      901 gaatatgaga gtatgaaata tttttggaag aaaaattctg ggaggaaggg tcttttctga
      961 gaaagtgtca ttaaatagga aaagtataat ttcccttttg agctagaatt gtatttgcaa
     1021 ttagaagctt ctcgttgcag aagtcgttct ttgacattat tctttgttcc ttaagatttc
     1081 catgaggaca acttacaagg acctgtatgg cctcatttta gctgcttcag tgtgttggag
     1141 ctgttacttg ctccttcatt tcctgggcct ggtgtgacag tggctactga gcgccatgga
     1201 ctacaaggac gacgatgaca aggtaccagg atctggggat tataaagatg atgatgataa
     1261 aggaggagga ggaagcggcg gaggcggaag cggaggagga ggaagcaagg agaagagtgc
     1321 ttgtcctaaa gatccagcca aacctccggc caaggcacaa gttgtgggat ggccaccggt
     1381 gagatcatac cggaagaacg tgatggtttc ctgccaaaaa tcaagcggtg gcccggaggc
     1441 ggcggcgttc gtgaaggtat caatggacgg agcaccgtac ttgaggaaaa tcgatttgag
     1501 gatgtataaa ggaggaggag gaagcggcgg aggcggaagc ggaggaggag gaagcatgaa
     1561 ggttgtccca gaaaagaatg ctgtccggat actctggggg cgagaacggg gtgctcgggc
     1621 catgggagct cagcggcttc tgcaggagct ggtagaagat aaaacccggt ggatgaaatg
     1681 ggagggcaag gtaaggaaaa tggctatatt tcacatagga tttgtgctgg tgatggtaga
     1741 atgagacctt agaatgggtt aacagcagag agttgggata tatttatctt gtgtgggaga
     1801 gttgacatat ctgacttagg gagttgtttg taaatttgaa agaactaatg tgctagaaat
     1861 atctttttcc tttagagcta gacagttttc ctgaaggtaa tttaggttta tctctctcaa
     1921 tttgtagctt gatcattgtc tccagttttt aagcctacac tgtgaatgct aatctagaat
     1981 ggttaatgtg cttttgagat acggtgaacc atggtctaca aatcatgtat tttaactttt
     2041 tttttttttt tgtctcccag agagtagaac tgccggatag tccacgctct accttcttat
     2101 tggccttcag cccagacagg tactcagtat ttatctataa cttaagattt gtgataacag
     2161 tgctttctgt atatttggcc tggcttgatc ttccaagtca gtccctaaag tctctctgaa
     2221 gtactgtatg aacagcactt tggccccgtg tcggaaaggg aaagaaagta gatgagcagt
     2281 gctaacctga aaaatcatga gaacaagcaa aggaaactga gaggatcaaa acttggaaag
     2341 tttctcaata acaatcattt gcctgtgtga tacagctcac cagtggtatg atggtttttc
     2401 cagtgggaaa aaatattttc cgaatgtggc atgtgtcatt tggggacagt gtggacaagc
     2461 caaaagcttt taaagatcag aatatttctg tttgtagata tactgcctaa catacaaagc
     2521 ttagtttctt tcatgttggg attggcaggt agcatcaata ttctatgtcg tccaaagttg
     2581 ataagattgg ttacccttcc acatgcttct gaattcagtt ctactctttg attttctagg
     2641 actctcttag cctccaccca tgtgaaccat aatatctata ttacggaggt gaagactggc
     2701 aagtgtgttc attccctgat tggacaccgc cgtactccat ggtgtgtcac ttttcatccc
     2761 accatctcag gccttattgc ttctggctgc ctagatgggg aggttaggat ttgggattta
     2821 cacgtgagta tttccttaag ataccc
//
