LOCUS Exported 3483 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Crispr repair template: add insertion site specific homology arms by PCR before insertion. ACCESSION . VERSION . KEYWORDS pJW2088 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 3483) AUTHORS Ashley G, Duong T, Levenson MT, Martinez MAQ, Johnsen LC, Hibshman JD, Saeger HN, Palmisano NJ, Doonan R, Martinez-Mendez R, Davidson B, 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 REFERENCE 2 (bases 1 to 3483) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..3483 /organism="synthetic DNA construct" /mol_type="other DNA" primer_bind 54..76 /label=M13/pUC Forward /note="In lacZ gene" primer_bind 68..85 /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind 69..85 /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" CDS 1071..1091 /codon_start=1 /product="tobacco etch virus (TEV) protease recognition and cleavage site" /label=TEV site /translation="ENLYFQS" CDS 1137..1202 /codon_start=1 /product="three tandem FLAG(R) epitope tags, followed by an enterokinase cleavage site" /label=3xFLAG /translation="DYKDHDGDYKDHDIDYKDDDDK" primer_bind complement(1256..1275) /label=T7 /note="T7 promoter, forward primer" promoter complement(1257..1275) /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" primer_bind complement(1280..1296) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(1280..1296) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(1398..1417) /label=pENTR-R /note="pENTR vectors, reverse primer" CDS 1409..2218 /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(1476..1495) /label=Kan-R /note="Kanamycin resistance gene, reverse primer" rep_origin 2365..2953 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 2854..2873 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 3107..3124 /label=L4440 /note="L4440 vector, forward primer" terminator 3283..3310 /label=rrnB T2 terminator /note="transcription terminator T2 from the E. coli rrnB gene" terminator 3402..5 /gene="Escherichia coli rrnB" /label=rrnB T1 terminator /note="transcription terminator T1 from the E. coli rrnB gene" ORIGIN 1 atttgatgcc tggcagttcc ctactctcgc gttaacgcta gcatggatgt tttcccagtc 61 acgacgttgt aaaacgacgg ccagtcttaa gctcgggccc cggtggcggt ggatcgggag 121 gaggaggttc gggtggcgga ggcagtggag gtggcggaag tggcggcggt ggttcaggag 181 gaggcggatc catgagtaaa ggagaagaat tgttcactgg agttgtccca atcctcgtcg 241 agctcgacgg agacgtcaac ggacacaagt tctccgtctc cggagaggga gagggagacg 301 ccacctacgg aaagctcacc ctcaagttca tctgcaccac cggaaagctc ccagtcccat 361 ggccaaccct cgtcaccacc ttctgctacg gagtccaatg cttctcccgt tacccagacc 421 acatgaagcg tcacgacttc ttcaagtccg ccatgccaga gggatacgtc caagagcgta 481 ccatcttctt caaggtaagt ttaaacatat atatactaac tactgattat ttaaattttc 541 aggacgacgg aaactacaag acccgtgccg aggtcaagtt cgagggagac accctcgtca 601 accgtatcga gctcaaggta agtttaaaca gttcggtact aactaaccat acatatttaa 661 attttcaggg aatcgacttc aaggaggacg gaaacatcct cggacacaag ctcgagtaca 721 actacaactc ccacaacgtc tacatcatgg ccgacaagca aaagaacgga atcaaggtca 781 acttcaaggt aagtttaaac atgattttac taactaacta atctgattta aattttcaga 841 tccgtcacaa catcgaggac ggatccgtcc aactcgccga ccactaccaa caaaacaccc 901 caatcggaga cggaccagtc ctcctcccag acaaccacta cctctccacc caatccgccc 961 tctccaagga cccaaacgag aagcgtgacc acatggtcct cctcgagttc gtcaccgccg 1021 ccggaatcac ccacggaatg gacgagctct acaagggcgg cagcggagga gagaatctgt 1081 actttcaatc cggatccgga ggtggcgggg gtggtggcgg ttctggctca ggatctgact 1141 acaaagacca tgacggtgat tataaagatc atgatatcga ttacaaggat gacgatgaca 1201 agggagcatc gggagcctca ggagcatcgt ccggaggcgc catccagctg atatccccta 1261 tagtgagtcg tattacatgg tcatagctgt ttcctggcag ctctggcccg tgtctcaaaa 1321 tctctgatgt tacattgcac aagataaaaa tatatcatca tgaacaataa aactgtctgc 1381 ttacataaac agtaatacaa ggggtgttat gagccatatt caacgggaaa cgtcgaggcc 1441 gcgattaaat tccaacatgg atgctgattt atatgggtat aaatgggctc gcgataatgt 1501 cgggcaatca ggtgcgacaa tctatcgctt gtatgggaag cccgatgcgc cagagttgtt 1561 tctgaaacat ggcaaaggta gcgttgccaa tgatgttaca gatgagatgg tcagactaaa 1621 ctggctgacg gaatttatgc ctcttccgac catcaagcat tttatccgta ctcctgatga 1681 tgcatggtta ctcaccactg cgatccccgg aaaaacagca ttccaggtat tagaagaata 1741 tcctgattca ggtgaaaata ttgttgatgc gctggcagtg ttcctgcgcc ggttgcattc 1801 gattcctgtt tgtaattgtc cttttaacag cgatcgcgta tttcgtctcg ctcaggcgca 1861 atcacgaatg aataacggtt tggttgatgc gagtgatttt gatgacgagc gtaatggctg 1921 gcctgttgaa caagtctgga aagaaatgca taaacttttg ccattctcac cggattcagt 1981 cgtcactcat ggtgatttct cacttgataa ccttattttt gacgagggga aattaatagg 2041 ttgtattgat gttggacgag tcggaatcgc agaccgatac caggatcttg ccatcctatg 2101 gaactgcctc ggtgagtttt ctccttcatt acagaaacgg ctttttcaaa aatatggtat 2161 tgataatcct gatatgaata aattgcagtt tcatttgatg ctcgatgagt ttttctaatc 2221 agaattggtt aattggttgt aacactggca gagcattacg ctgacttgac gggacggcgc 2281 aagctcatga ccaaaatccc ttaacgtgag ttacgcgtcg ttccactgag cgtcagaccc 2341 cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt 2401 gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac 2461 tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg ttcttctagt 2521 gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct 2581 gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga 2641 ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac 2701 acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg 2761 agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt 2821 cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc 2881 tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg 2941 gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc 3001 ttttgctcac atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc 3061 ctttgagtga gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag 3121 cgaggaagcg gaagagcgcc caatacgcaa accgcctctc cccgcgcgtt ggccgattca 3181 ttaatgcagc tggcacgaca ggtttcccga ctggaaagcg ggcagtgagc gcaacgcaat 3241 taatacgcgt accgctagcc aggaagagtt tgtagaaacg caaaaaggcc atccgtcagg 3301 atggccttct gcttagtttg atgcctggca gtttatggcg ggcgtcctgc ccgccaccct 3361 ccgggccgtt gcttcacaac gttcaaatcc gctcccggcg gatttgtcct actcaggaga 3421 gcgttcaccg acaaacaaca gataaaacga aaggcccagt cttccgactg agcctttcgt 3481 ttt //