LOCUS       sequence-320582-        3603 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  CRISPR repair template: add insertion site specific homology arms by
            PCR before insertion.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3603)
  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 3603)
  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: pJW2098
FEATURES             Location/Qualifiers
     source          1..3603
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      complement(62..148)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     197..219
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     211..228
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     212..228
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             1199..1219
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             1241..1372
                     /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             1400..1465
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     primer_bind     complement(1519..1538)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1520..1538)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1543..1559)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1543..1559)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1661..1680)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             1672..2481
                     /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     complement(1739..1758)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      2628..3216
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3117..3136
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3370..3387
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(3546..3573)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
ORIGIN
        1 ggcgtcctgc ccgccaccct ccgggccgtt gcttcacaac gttcaaatcc gctcccggcg
       61 gatttgtcct actcaggaga gcgttcaccg acaaacaaca gataaaacga aaggcccagt
      121 cttccgactg agcctttcgt tttatttgat gcctggcagt tccctactct cgcgttaacg
      181 ctagcatgga tgttttccca gtcacgacgt tgtaaaacga cggccagtct taagctcggg
      241 ccccggtggc ggtggatcgg gaggaggagg ttcgggtggc ggaggcagcg gaggaggcgg
      301 aagtggcggc ggtggttcag gaggaggcgg atccatggtc tccaagggag aggctgtcat
      361 caaggagttc atgcgtttca aggtccacat ggagggatcc atgaacggac acgagttcga
      421 gatcgaggga gagggagagg gacgtccata cgagggaacc caaaccgcca agctcaaggt
      481 caccaaggga ggtaagtttg tgataatcca atttcaattc gcaatggtca tcgttttttc
      541 aggaccactc ccattctcct gggacatcct ctccccacaa ttcatgtacg gatcccgtgc
      601 cttcatcaag cacccagccg acatcccaga ctactacaag caatccttcc cagagggatt
      661 caagtgggag cgtgtgatga acttcgaggt aagtttttac tccgctttta acaatggttg
      721 tttgacatca ttttttcagg acggaggagc cgtcaccgtc acccaagaca cctccctcga
      781 ggacggaacc ctcatctaca aggtcaagct ccgtggaacc aacttcccac cagacggacc
      841 agtcatgcaa aagaagacca tgggatggga ggcctccacc gagcgtctct acccagagga
      901 cggagtcctc aaaggtaagt ttgatgaaac ggtttcgtct tatatacact aatggtactt
      961 ttcaggagac atcaagatgg ccctccgtct caaggacgga ggacgttacc tcgccgactt
     1021 caagaccacc tacaaggcca agaagccagt ccaaatgcca ggagcctaca acgtcgaccg
     1081 taagctcgac atcacctccc acaacgagga ctacaccgtc gtcgagcaat acgagcgttc
     1141 cgagggacgt cactccaccg gaggaatgga cgagctctac aagggcggca gcggaggaga
     1201 gaatctgtac tttcaatccg gatccggagg tggcgggatg cctaaagatc cagccaaacc
     1261 tccggccaag gcacaagttg tgggatggcc accggtgaga tcataccgga agaacgtgat
     1321 ggtttcctgc caaaaatcaa gcggtggccc ggaggcggcg gcgttcgtga agggtggtgg
     1381 cggttctggc tcaggatctg actacaaaga ccatgacggt gattataaag atcatgatat
     1441 cgattacaag gatgacgatg acaagggagc atcgggagcc tcaggagcat cgtccggagg
     1501 cgccatccag ctgatatccc ctatagtgag tcgtattaca tggtcatagc tgtttcctgg
     1561 cagctctggc ccgtgtctca aaatctctga tgttacattg cacaagataa aaatatatca
     1621 tcatgaacaa taaaactgtc tgcttacata aacagtaata caaggggtgt tatgagccat
     1681 attcaacggg aaacgtcgag gccgcgatta aattccaaca tggatgctga tttatatggg
     1741 tataaatggg ctcgcgataa tgtcgggcaa tcaggtgcga caatctatcg cttgtatggg
     1801 aagcccgatg cgccagagtt gtttctgaaa catggcaaag gtagcgttgc caatgatgtt
     1861 acagatgaga tggtcagact aaactggctg acggaattta tgcctcttcc gaccatcaag
     1921 cattttatcc gtactcctga tgatgcatgg ttactcacca ctgcgatccc cggaaaaaca
     1981 gcattccagg tattagaaga atatcctgat tcaggtgaaa atattgttga tgcgctggca
     2041 gtgttcctgc gccggttgca ttcgattcct gtttgtaatt gtccttttaa cagcgatcgc
     2101 gtatttcgtc tcgctcaggc gcaatcacga atgaataacg gtttggttga tgcgagtgat
     2161 tttgatgacg agcgtaatgg ctggcctgtt gaacaagtct ggaaagaaat gcataaactt
     2221 ttgccattct caccggattc agtcgtcact catggtgatt tctcacttga taaccttatt
     2281 tttgacgagg ggaaattaat aggttgtatt gatgttggac gagtcggaat cgcagaccga
     2341 taccaggatc ttgccatcct atggaactgc ctcggtgagt tttctccttc attacagaaa
     2401 cggctttttc aaaaatatgg tattgataat cctgatatga ataaattgca gtttcatttg
     2461 atgctcgatg agtttttcta atcagaattg gttaattggt tgtaacactg gcagagcatt
     2521 acgctgactt gacgggacgg cgcaagctca tgaccaaaat cccttaacgt gagttacgcg
     2581 tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt
     2641 tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt
     2701 ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag
     2761 ataccaaata ctgttcttct agtgtagccg tagttaggcc accacttcaa gaactctgta
     2821 gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat
     2881 aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg
     2941 ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg
     3001 agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac
     3061 aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga
     3121 aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt
     3181 ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta
     3241 cggttcctgg ccttttgctg gccttttgct cacatgttct ttcctgcgtt atcccctgat
     3301 tctgtggata accgtattac cgcctttgag tgagctgata ccgctcgccg cagccgaacg
     3361 accgagcgca gcgagtcagt gagcgaggaa gcggaagagc gcccaatacg caaaccgcct
     3421 ctccccgcgc gttggccgat tcattaatgc agctggcacg acaggtttcc cgactggaaa
     3481 gcgggcagtg agcgcaacgc aattaatacg cgtaccgcta gccaggaaga gtttgtagaa
     3541 acgcaaaaag gccatccgtc aggatggcct tctgcttagt ttgatgcctg gcagtttatg
     3601 gcg
//
