LOCUS       sequence-76761-e         906 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  for Cre-lox cassette exchange of C-terminal HA-auxin-inducible 
            degron tagged gene sequences into the S. pombe urg1 locus.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 906)
  AUTHORS   Watson AT, Daigaku Y, Mohebi S, Etheridge TJ, Chahwan C, Murray JM, 
            Carr AM
  TITLE     Optimisation of the Schizosaccharomyces pombe urg1 Expression 
            System.
  JOURNAL   PLoS One. 2013 Dec 20;8(12):e83800. doi: 
            10.1371/journal.pone.0083800.
  PUBMED    24376751
REFERENCE   2  (bases 1 to 906)
  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: "PLoS One.";
            date: "2013-12-20"; volume: "8(12):e83800. doi"; pages: " 
            10.1371/journal.pone.0083800"
COMMENT     Sequence Label: pAW8ENdeI-cHAIAA17
FEATURES             Location/Qualifiers
     source          1..906
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     protein_bind    114..135
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(150..155,156..173,174..180)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    188..204
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     193..215
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     212..228
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     212..228
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     protein_bind    complement(247..280)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     CDS             369..395
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             642..845
                     /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 gcggaagagc gcccaatacg caaaccgcct ctccccgcgc gttggccgat tcattaatgc
       61 agctggcacg acaggtttcc cgactggaaa gcgggcagtg agcgcaacgc aattaatgtg
      121 agttagctca ctcattaggc accccaggct ttacacttta tgcttccggc tcgtatgttg
      181 tgtggaattg tgagcggata acaatttcac acaggaaaca gctatgacca tgattacgcc
      241 aagcttataa cttcgtatag catacattat acgaagttat tgctagccgt tttcttcaat
      301 tactaaatta gaactaattc aatcatatgc tggcatgcgg agagctcgga gtcgacgaga
      361 ctagtggtta tccttatgat gttcctgatt atgcttctag tactggttct acaggatcaa
      421 ccggctccac ggggtcgatg ggtagatcta tgatgggcag tgtcgagctg aatctgaggg
      481 agactgagct gtgtcttggt cttcccggtg gagatacagt ggctccggta accggaaaca
      541 agagagggtt ctcagagacg gttgatctga agctaaatct gaataatgag cctgcaaaca
      601 aggaaggatc tacgactcat gacgtcgtga cttttgattc caaggagaag agtgcttgtc
      661 ctaaagatcc agccaaacct ccggccaagg cacaagttgt gggatggcca ccggtgagat
      721 cataccggaa gaacgtgatg gtttcctgcc aaaaatcaag cggtggcccg gaggcggcgg
      781 cgttcgtgaa ggtatcaatg gacggagcac cgtacttgag gaaaatcgat ttgaggatgt
      841 ataaaagcta cgatgagctt tctaatgctt tgtccaacat gttcagctct tttaccatgg
      901 gcaaac
//
