LOCUS Exported 3423 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 pJW2072 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 3423) 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 3423) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..3423 /organism="synthetic DNA construct" /mol_type="other DNA" terminator 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 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 //