LOCUS       sequence-309451-        2949 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  NT Donor-cassette for SapTrap.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2949)
  AUTHORS   Ashley GE, Duong T, Levenson MT, Martinez MAQ, Johnson LC, Hibshman 
            JD, Saeger HN, Palmisano NJ, Doonan R, Martinez-Mendez R, Davidson 
            BR, Zhang W, Ragle JM, Medwig-Kinney TN, Sirota SS, Goldstein B, 
            Matus DQ, Dickinson DJ, Reiner DJ, Ward JD
  TITLE     An expanded auxin-inducible degron toolkit for Caenorhabditis 
            elegans.
  JOURNAL   Genetics. 2021 Mar 31;217(3). pii: 6104563. doi: 
            10.1093/genetics/iyab006.
  PUBMED    33677541
REFERENCE   2  (bases 1 to 2949)
  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: "Genetics.";
            date: "2021-03-31"; volume: "217(3). pii: 6104563. doi"; pages: " 
            10.1093/genetics/iyab006"
COMMENT     Sequence Label: pJW1787
FEATURES             Location/Qualifiers
     source          1..2949
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             33..53
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             171..302
                     /codon_start=1
                     /product="minimal degron from the auxin-responsive IAA17 
                     protein of Arabidopsis thaliana (Morawska and Ulrich, 
                     2013)"
                     /label=AID*
                     /note="acts as an auxin-inducible degron (AID) in the 
                     presence of TIR1"
                     /translation="PKDPAKPPAKAQVVGWPPVRSYRKNVMVSCQKSSGGPEAAAFVK"
     CDS             330..395
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     protein_bind    complement(408..507)
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(523..540)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(523..539)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(532..554)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     terminator      603..689
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      781..808
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     primer_bind     complement(967..984)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1138..1726)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1218..1237)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(1873..2682)
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP
                     DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA
                     FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
                     FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
                     RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     2596..2615
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     2674..2693
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     primer_bind     2795..2811
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2795..2811
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     2816..2835
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2816..2834
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    join(2852..2949,1..2)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
ORIGIN
        1 gtcctagctc ttcgggtggc ggcagcggag gagagaatct gtactttcaa tccggatccg
       61 gaggtggcgg gatggcttct tctcttcgtc agatcctcga ctctcagaag atggagtggc
      121 gttctaacgc tggaggatct ggcggaggag gcagtggtag tggcagtatg cctaaagatc
      181 cagccaaacc tccggccaag gcacaagttg tgggatggcc accggtgaga tcataccgga
      241 agaacgtgat ggtttcctgc caaaaatcaa gcggtggccc ggaggcggcg gcgttcgtga
      301 agggtggtgg cggttctggc tcaggatctg actacaaaga ccatgacggt gattataaag
      361 atcatgatat cgattacaag gatgacgatg acaagacgcg aagagcgacc cagctttttt
      421 gtacaaagtt ggcattataa aaaagcattg ctcatcaatt tgttgcaacg aacaggtcac
      481 tatcagtcaa aataaaatca ttatttgggg cccgagctta agactggccg tcgttttaca
      541 acgtcgtgac tgggaaaaca tccatgctag cgttaacgcg agagtaggga actgccaggc
      601 atcaaataaa acgaaaggct cagtcggaag actgggcctt tcgttttatc tgttgtttgt
      661 cggtgaacgc tctcctgagt aggacaaatc cgccgggagc ggatttgaac gttgtgaagc
      721 aacggcccgg agggtggcgg gcaggacgcc cgccataaac tgccaggcat caaactaagc
      781 agaaggccat cctgacggat ggcctttttg cgtttctaca aactcttcct ggctagcggt
      841 acgcgtatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca
      901 gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc
      961 cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc
     1021 tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat
     1081 gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt
     1141 ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg
     1201 aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc
     1261 tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt
     1321 ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa
     1381 gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta
     1441 tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa
     1501 caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa
     1561 ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt
     1621 cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt
     1681 ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat
     1741 cttttctacg gggtctgacg ctcagtggaa cgacgcgtaa ctcacgttaa gggattttgg
     1801 tcatgagctt gcgccgtccc gtcaagtcag cgtaatgctc tgccagtgtt acaaccaatt
     1861 aaccaattct gattagaaaa actcatcgag catcaaatga aactgcaatt tattcatatc
     1921 aggattatca ataccatatt tttgaaaaag ccgtttctgt aatgaaggag aaaactcacc
     1981 gaggcagttc cataggatgg caagatcctg gtatcggtct gcgattccga ctcgtccaac
     2041 atcaatacaa cctattaatt tcccctcgtc aaaaataagg ttatcaagtg agaaatcacc
     2101 atgagtgacg actgaatccg gtgagaatgg caaaagttta tgcatttctt tccagacttg
     2161 ttcaacaggc cagccattac gctcgtcatc aaaatcactc gcatcaacca aaccgttatt
     2221 cattcgtgat tgcgcctgag cgagacgaaa tacgcgatcg ctgttaaaag gacaattaca
     2281 aacaggaatc gaatgcaacc ggcgcaggaa cactgccagc gcatcaacaa tattttcacc
     2341 tgaatcagga tattcttcta atacctggaa tgctgttttt ccggggatcg cagtggtgag
     2401 taaccatgca tcatcaggag tacggataaa atgcttgatg gtcggaagag gcataaattc
     2461 cgtcagccag tttagtctga ccatctcatc tgtaacatca ttggcaacgc tacctttgcc
     2521 atgtttcaga aacaactctg gcgcatcggg cttcccatac aagcgataga ttgtcgcacc
     2581 tgattgcccg acattatcgc gagcccattt atacccatat aaatcagcat ccatgttgga
     2641 atttaatcgc ggcctcgacg tttcccgttg aatatggctc ataacacccc ttgtattact
     2701 gtttatgtaa gcagacagtt ttattgttca tgatgatata tttttatctt gtgcaatgta
     2761 acatcagaga ttttgagaca cgggccagag ctgccaggaa acagctatga ccatgtaata
     2821 cgactcacta taggggatat cagctggatg gcaaataatg attttatttt gactgatagt
     2881 gacctgttcg ttgcaacaaa ttgataagca atgctttctt ataatgccaa ctttgtacaa
     2941 gaaagctgg
//
