LOCUS       sequence-309696-        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"
     primer_bind     1..17
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1..17
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     22..41
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        22..40
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    58..157
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             188..208
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             326..457
                     /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             485..550
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     protein_bind    complement(563..662)
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(678..695)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(678..694)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(687..709)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     terminator      758..844
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      936..963
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     primer_bind     complement(1122..1139)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1293..1881)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1373..1392)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(2028..2837)
                     /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     2751..2770
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     2829..2848
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
ORIGIN
        1 caggaaacag ctatgaccat gtaatacgac tcactatagg ggatatcagc tggatggcaa
       61 ataatgattt tattttgact gatagtgacc tgttcgttgc aacaaattga taagcaatgc
      121 tttcttataa tgccaacttt gtacaagaaa gctgggtcct agctcttcgg gtggcggcag
      181 cggaggagag aatctgtact ttcaatccgg atccggaggt ggcgggatgg cttcttctct
      241 tcgtcagatc ctcgactctc agaagatgga gtggcgttct aacgctggag gatctggcgg
      301 aggaggcagt ggtagtggca gtatgcctaa agatccagcc aaacctccgg ccaaggcaca
      361 agttgtggga tggccaccgg tgagatcata ccggaagaac gtgatggttt cctgccaaaa
      421 atcaagcggt ggcccggagg cggcggcgtt cgtgaagggt ggtggcggtt ctggctcagg
      481 atctgactac aaagaccatg acggtgatta taaagatcat gatatcgatt acaaggatga
      541 cgatgacaag acgcgaagag cgacccagct tttttgtaca aagttggcat tataaaaaag
      601 cattgctcat caatttgttg caacgaacag gtcactatca gtcaaaataa aatcattatt
      661 tggggcccga gcttaagact ggccgtcgtt ttacaacgtc gtgactggga aaacatccat
      721 gctagcgtta acgcgagagt agggaactgc caggcatcaa ataaaacgaa aggctcagtc
      781 ggaagactgg gcctttcgtt ttatctgttg tttgtcggtg aacgctctcc tgagtaggac
      841 aaatccgccg ggagcggatt tgaacgttgt gaagcaacgg cccggagggt ggcgggcagg
      901 acgcccgcca taaactgcca ggcatcaaac taagcagaag gccatcctga cggatggcct
      961 ttttgcgttt ctacaaactc ttcctggcta gcggtacgcg tattaattgc gttgcgctca
     1021 ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc
     1081 gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg
     1141 cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta
     1201 tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc
     1261 aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag
     1321 catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac
     1381 caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc
     1441 ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt
     1501 aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc
     1561 gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga
     1621 cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta
     1681 ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta
     1741 tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga
     1801 tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg
     1861 cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag
     1921 tggaacgacg cgtaactcac gttaagggat tttggtcatg agcttgcgcc gtcccgtcaa
     1981 gtcagcgtaa tgctctgcca gtgttacaac caattaacca attctgatta gaaaaactca
     2041 tcgagcatca aatgaaactg caatttattc atatcaggat tatcaatacc atatttttga
     2101 aaaagccgtt tctgtaatga aggagaaaac tcaccgaggc agttccatag gatggcaaga
     2161 tcctggtatc ggtctgcgat tccgactcgt ccaacatcaa tacaacctat taatttcccc
     2221 tcgtcaaaaa taaggttatc aagtgagaaa tcaccatgag tgacgactga atccggtgag
     2281 aatggcaaaa gtttatgcat ttctttccag acttgttcaa caggccagcc attacgctcg
     2341 tcatcaaaat cactcgcatc aaccaaaccg ttattcattc gtgattgcgc ctgagcgaga
     2401 cgaaatacgc gatcgctgtt aaaaggacaa ttacaaacag gaatcgaatg caaccggcgc
     2461 aggaacactg ccagcgcatc aacaatattt tcacctgaat caggatattc ttctaatacc
     2521 tggaatgctg tttttccggg gatcgcagtg gtgagtaacc atgcatcatc aggagtacgg
     2581 ataaaatgct tgatggtcgg aagaggcata aattccgtca gccagtttag tctgaccatc
     2641 tcatctgtaa catcattggc aacgctacct ttgccatgtt tcagaaacaa ctctggcgca
     2701 tcgggcttcc catacaagcg atagattgtc gcacctgatt gcccgacatt atcgcgagcc
     2761 catttatacc catataaatc agcatccatg ttggaattta atcgcggcct cgacgtttcc
     2821 cgttgaatat ggctcataac accccttgta ttactgttta tgtaagcaga cagttttatt
     2881 gttcatgatg atatattttt atcttgtgca atgtaacatc agagattttg agacacgggc
     2941 cagagctgc
//
