LOCUS       sequence-318659-        3423 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 3423)
  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 3423)
  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: pJW2072
FEATURES             Location/Qualifiers
     source          1..3423
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      complement(2..88)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     137..159
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     151..168
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     152..168
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             1136..1225
                     /codon_start=1
                     /product="3 tandem Myc epitope tags"
                     /label=3xMyc
                     /translation="EQKLISEEDLEQKLISEEDLEQKLISEEDL"
     primer_bind     complement(1279..1298)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1280..1298)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1303..1319)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1303..1319)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1421..1440)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             1432..2241
                     /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(1499..1518)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      2388..2976
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2877..2896
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3130..3147
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(3306..3333)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
ORIGIN
        1 gatttgtcct actcaggaga gcgttcaccg acaaacaaca gataaaacga aaggcccagt
       61 cttccgactg agcctttcgt tttatttgat gcctggcagt tccctactct cgcgttaacg
      121 ctagcatgga tgttttccca gtcacgacgt tgtaaaacga cggccagtct taagctcggg
      181 ccccggtggc ggtggatcgg gaggaggagg ttcgggtggc ggaggcagtg gaggtggcgg
      241 aagtggcggc ggtggttcag gaggaggcgg atccatggtc tccaaaggag aagctgtgat
      301 caaggagttc atgcgcttca aggttcacat ggaaggaagc atgaatggtc acgagttcga
      361 aatcgaagga gaaggggagg gccgcccgta cgagggaact cagaccgcca agctgaaggt
      421 caccaaggga ggtaagtttg tgataatcca atttcaattc gcaatggtca tcgttttttc
      481 aggaccactt ccattctcat gggatattct ctccccacaa ttcatgtacg gctcccgtgc
      541 tttcatcaaa cacccagccg acattccaga ttactacaag caatctttcc cagaaggctt
      601 caagtgggag cgtgtgatga acttcgagga tggtggagca gttacagtaa ctcaggacac
      661 ttctctcgag gtaagttttt actccgcttt taacaatggt tgtttgacat cattttttca
      721 ggatggcact ttgatctaca aggtcaagct ccgtggtacc aatttcccac cagatggacc
      781 agttatgcag aagaagacga tgggatggga ggcttccacc gaacgattgt acccagaaga
      841 tggagttctc aagggagaca tcaaaatggc tcttcgcctc aaggacggag gtaagtttga
      901 tgaaacggtt tcgtcttata tacactaatg gtacttttca ggacgttacc tggcggattt
      961 caagaccaca tacaaggcaa agaagccagt tcaaatgcca ggagcatata acgttgaccg
     1021 caagcttgat attacttccc ataatgaaga ctacacagtt gtagaacaat acgaacgatc
     1081 cgagggacgt cattcgaccg gaggaatgga cgagctgtac aagggagccg gatctgaaca
     1141 aaaacttatt tctgaagagg atcttgagca aaagctcatc tccgaggagg acctcgagca
     1201 gaagttgatc agcgaggaag acttgggagc atcgggagcc tcaggagcat cgtccggagg
     1261 cgccatccag ctgatatccc ctatagtgag tcgtattaca tggtcatagc tgtttcctgg
     1321 cagctctggc ccgtgtctca aaatctctga tgttacattg cacaagataa aaatatatca
     1381 tcatgaacaa taaaactgtc tgcttacata aacagtaata caaggggtgt tatgagccat
     1441 attcaacggg aaacgtcgag gccgcgatta aattccaaca tggatgctga tttatatggg
     1501 tataaatggg ctcgcgataa tgtcgggcaa tcaggtgcga caatctatcg cttgtatggg
     1561 aagcccgatg cgccagagtt gtttctgaaa catggcaaag gtagcgttgc caatgatgtt
     1621 acagatgaga tggtcagact aaactggctg acggaattta tgcctcttcc gaccatcaag
     1681 cattttatcc gtactcctga tgatgcatgg ttactcacca ctgcgatccc cggaaaaaca
     1741 gcattccagg tattagaaga atatcctgat tcaggtgaaa atattgttga tgcgctggca
     1801 gtgttcctgc gccggttgca ttcgattcct gtttgtaatt gtccttttaa cagcgatcgc
     1861 gtatttcgtc tcgctcaggc gcaatcacga atgaataacg gtttggttga tgcgagtgat
     1921 tttgatgacg agcgtaatgg ctggcctgtt gaacaagtct ggaaagaaat gcataaactt
     1981 ttgccattct caccggattc agtcgtcact catggtgatt tctcacttga taaccttatt
     2041 tttgacgagg ggaaattaat aggttgtatt gatgttggac gagtcggaat cgcagaccga
     2101 taccaggatc ttgccatcct atggaactgc ctcggtgagt tttctccttc attacagaaa
     2161 cggctttttc aaaaatatgg tattgataat cctgatatga ataaattgca gtttcatttg
     2221 atgctcgatg agtttttcta atcagaattg gttaattggt tgtaacactg gcagagcatt
     2281 acgctgactt gacgggacgg cgcaagctca tgaccaaaat cccttaacgt gagttacgcg
     2341 tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt
     2401 tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt
     2461 ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag
     2521 ataccaaata ctgttcttct agtgtagccg tagttaggcc accacttcaa gaactctgta
     2581 gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat
     2641 aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg
     2701 ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg
     2761 agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac
     2821 aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga
     2881 aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt
     2941 ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta
     3001 cggttcctgg ccttttgctg gccttttgct cacatgttct ttcctgcgtt atcccctgat
     3061 tctgtggata accgtattac cgcctttgag tgagctgata ccgctcgccg cagccgaacg
     3121 accgagcgca gcgagtcagt gagcgaggaa gcggaagagc gcccaatacg caaaccgcct
     3181 ctccccgcgc gttggccgat tcattaatgc agctggcacg acaggtttcc cgactggaaa
     3241 gcgggcagtg agcgcaacgc aattaatacg cgtaccgcta gccaggaaga gtttgtagaa
     3301 acgcaaaaag gccatccgtc aggatggcct tctgcttagt ttgatgcctg gcagtttatg
     3361 gcgggcgtcc tgcccgccac cctccgggcc gttgcttcac aacgttcaaa tccgctcccg
     3421 gcg
//
