LOCUS       sequence-320769-        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"
     primer_bind     92..109
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(268..295)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(387..473)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     522..544
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     536..553
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     537..553
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             1524..1544
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             1566..1697
                     /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             1725..1790
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     primer_bind     complement(1844..1863)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1845..1863)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1868..1884)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1868..1884)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1986..2005)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             1997..2806
                     /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(2064..2083)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      2953..3541
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3442..3461
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga
       61 taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga
      121 gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctggca
      181 cgacaggttt cccgactgga aagcgggcag tgagcgcaac gcaattaata cgcgtaccgc
      241 tagccaggaa gagtttgtag aaacgcaaaa aggccatccg tcaggatggc cttctgctta
      301 gtttgatgcc tggcagttta tggcgggcgt cctgcccgcc accctccggg ccgttgcttc
      361 acaacgttca aatccgctcc cggcggattt gtcctactca ggagagcgtt caccgacaaa
      421 caacagataa aacgaaaggc ccagtcttcc gactgagcct ttcgttttat ttgatgcctg
      481 gcagttccct actctcgcgt taacgctagc atggatgttt tcccagtcac gacgttgtaa
      541 aacgacggcc agtcttaagc tcgggccccg gtggcggtgg atcgggagga ggaggttcgg
      601 gtggcggagg cagcggagga ggcggaagtg gcggcggtgg ttcaggagga ggcggatcca
      661 tggtctccaa gggagaggct gtcatcaagg agttcatgcg tttcaaggtc cacatggagg
      721 gatccatgaa cggacacgag ttcgagatcg agggagaggg agagggacgt ccatacgagg
      781 gaacccaaac cgccaagctc aaggtcacca agggaggtaa gtttgtgata atccaatttc
      841 aattcgcaat ggtcatcgtt ttttcaggac cactcccatt ctcctgggac atcctctccc
      901 cacaattcat gtacggatcc cgtgccttca tcaagcaccc agccgacatc ccagactact
      961 acaagcaatc cttcccagag ggattcaagt gggagcgtgt gatgaacttc gaggtaagtt
     1021 tttactccgc ttttaacaat ggttgtttga catcattttt tcaggacgga ggagccgtca
     1081 ccgtcaccca agacacctcc ctcgaggacg gaaccctcat ctacaaggtc aagctccgtg
     1141 gaaccaactt cccaccagac ggaccagtca tgcaaaagaa gaccatggga tgggaggcct
     1201 ccaccgagcg tctctaccca gaggacggag tcctcaaagg taagtttgat gaaacggttt
     1261 cgtcttatat acactaatgg tacttttcag gagacatcaa gatggccctc cgtctcaagg
     1321 acggaggacg ttacctcgcc gacttcaaga ccacctacaa ggccaagaag ccagtccaaa
     1381 tgccaggagc ctacaacgtc gaccgtaagc tcgacatcac ctcccacaac gaggactaca
     1441 ccgtcgtcga gcaatacgag cgttccgagg gacgtcactc caccggagga atggacgagc
     1501 tctacaaggg cggcagcgga ggagagaatc tgtactttca atccggatcc ggaggtggcg
     1561 ggatgcctaa agatccagcc aaacctccgg ccaaggcaca agttgtggga tggccaccgg
     1621 tgagatcata ccggaagaac gtgatggttt cctgccaaaa atcaagcggt ggcccggagg
     1681 cggcggcgtt cgtgaagggt ggtggcggtt ctggctcagg atctgactac aaagaccatg
     1741 acggtgatta taaagatcat gatatcgatt acaaggatga cgatgacaag ggagcatcgg
     1801 gagcctcagg agcatcgtcc ggaggcgcca tccagctgat atcccctata gtgagtcgta
     1861 ttacatggtc atagctgttt cctggcagct ctggcccgtg tctcaaaatc tctgatgtta
     1921 cattgcacaa gataaaaata tatcatcatg aacaataaaa ctgtctgctt acataaacag
     1981 taatacaagg ggtgttatga gccatattca acgggaaacg tcgaggccgc gattaaattc
     2041 caacatggat gctgatttat atgggtataa atgggctcgc gataatgtcg ggcaatcagg
     2101 tgcgacaatc tatcgcttgt atgggaagcc cgatgcgcca gagttgtttc tgaaacatgg
     2161 caaaggtagc gttgccaatg atgttacaga tgagatggtc agactaaact ggctgacgga
     2221 atttatgcct cttccgacca tcaagcattt tatccgtact cctgatgatg catggttact
     2281 caccactgcg atccccggaa aaacagcatt ccaggtatta gaagaatatc ctgattcagg
     2341 tgaaaatatt gttgatgcgc tggcagtgtt cctgcgccgg ttgcattcga ttcctgtttg
     2401 taattgtcct tttaacagcg atcgcgtatt tcgtctcgct caggcgcaat cacgaatgaa
     2461 taacggtttg gttgatgcga gtgattttga tgacgagcgt aatggctggc ctgttgaaca
     2521 agtctggaaa gaaatgcata aacttttgcc attctcaccg gattcagtcg tcactcatgg
     2581 tgatttctca cttgataacc ttatttttga cgaggggaaa ttaataggtt gtattgatgt
     2641 tggacgagtc ggaatcgcag accgatacca ggatcttgcc atcctatgga actgcctcgg
     2701 tgagttttct ccttcattac agaaacggct ttttcaaaaa tatggtattg ataatcctga
     2761 tatgaataaa ttgcagtttc atttgatgct cgatgagttt ttctaatcag aattggttaa
     2821 ttggttgtaa cactggcaga gcattacgct gacttgacgg gacggcgcaa gctcatgacc
     2881 aaaatccctt aacgtgagtt acgcgtcgtt ccactgagcg tcagaccccg tagaaaagat
     2941 caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa
     3001 accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa
     3061 ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt
     3121 aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt
     3181 accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata
     3241 gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt
     3301 ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac
     3361 gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga
     3421 gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg
     3481 ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa
     3541 aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat
     3601 gtt
//
