LOCUS Exported 2949 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION CT Donor-cassette for SapTrap. ACCESSION . VERSION . KEYWORDS pJW1788 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 2949) 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 2949) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..2949 /organism="synthetic DNA construct" /mol_type="other DNA" primer_bind 1..17 /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind 1..17 /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind 22..41 /label=T7 /note="T7 promoter, forward primer" promoter 22..40 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" protein_bind 58..157 /gene="mutant version of attL" /label=attL2 /bound_moiety="LR Clonase(TM)" /note="recombination site for the Gateway(R) LR reaction" CDS 188..208 /codon_start=1 /product="tobacco etch virus (TEV) protease recognition and cleavage site" /label=TEV site /translation="ENLYFQS" CDS 485..550 /codon_start=1 /product="three tandem FLAG(R) epitope tags, followed by an enterokinase cleavage site" /label=3xFLAG /translation="DYKDHDGDYKDHDIDYKDDDDK" protein_bind complement(563..662) /gene="mutant version of attL" /label=attL1 /bound_moiety="LR Clonase(TM)" /note="recombination site for the Gateway(R) LR reaction" primer_bind complement(678..695) /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind complement(678..694) /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" primer_bind complement(687..709) /label=M13/pUC Forward /note="In lacZ gene" terminator 758..844 /gene="Escherichia coli rrnB" /label=rrnB T1 terminator /note="transcription terminator T1 from the E. coli rrnB gene" terminator 936..963 /label=rrnB T2 terminator /note="transcription terminator T2 from the E. coli rrnB gene" primer_bind complement(1122..1139) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(1293..1881) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(1373..1392) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(2028..2837) /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 2751..2770 /label=Kan-R /note="Kanamycin resistance gene, reverse primer" primer_bind 2829..2848 /label=pENTR-R /note="pENTR vectors, reverse primer" ORIGIN 1 caggaaacag ctatgaccat gtaatacgac tcactatagg ggatatcagc tggatggcaa 61 ataatgattt tattttgact gatagtgacc tgttcgttgc aacaaattga taagcaatgc 121 tttcttataa tgccaacttt gtacaagaaa gctgggtcct agctcttcgg cgggcggcag 181 cggaggagag aatctgtact ttcaatccgg atccggaggt ggcgggatgg cttcttctct 241 tcgtcagatc ctcgactctc agaagatgga gtggcgttct aacgctggag gatctggcgg 301 aggaggcagt ggtagtggca gtatgcctaa agatccagcc aaacctccgg ccaaggcaca 361 agttgtggga tggccaccgg tgagatcata ccggaagaac gtgatggttt cctgccaaaa 421 atcaagcggt ggcccggagg cggcggcgtt cgtgaagggt ggtggcggtt ctggctcagg 481 atctgactac aaagaccatg acggtgatta taaagatcat gatatcgatt acaaggatga 541 cgatgacaag atgcgaagag cgacccagct tttttgtaca aagttggcat tataaaaaag 601 cattgctcat caatttgttg caacgaacag gtcactatca gtcaaaataa aatcattatt 661 tggggcccga gcttaagact ggccgtcgtt ttacaacgtc gtgactggga aaacatccat 721 gctagcgtta acgcgagagt agggaactgc caggcatcaa ataaaacgaa aggctcagtc 781 ggaagactgg gcctttcgtt ttatctgttg tttgtcggtg aacgctctcc tgagtaggac 841 aaatccgccg ggagcggatt tgaacgttgt gaagcaacgg cccggagggt ggcgggcagg 901 acgcccgcca taaactgcca ggcatcaaac taagcagaag gccatcctga cggatggcct 961 ttttgcgttt ctacaaactc ttcctggcta gcggtacgcg tattaattgc gttgcgctca 1021 ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc 1081 gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg 1141 cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta 1201 tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc 1261 aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag 1321 catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac 1381 caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc 1441 ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt 1501 aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc 1561 gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga 1621 cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta 1681 ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta 1741 tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga 1801 tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg 1861 cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag 1921 tggaacgacg cgtaactcac gttaagggat tttggtcatg agcttgcgcc gtcccgtcaa 1981 gtcagcgtaa tgctctgcca gtgttacaac caattaacca attctgatta gaaaaactca 2041 tcgagcatca aatgaaactg caatttattc atatcaggat tatcaatacc atatttttga 2101 aaaagccgtt tctgtaatga aggagaaaac tcaccgaggc agttccatag gatggcaaga 2161 tcctggtatc ggtctgcgat tccgactcgt ccaacatcaa tacaacctat taatttcccc 2221 tcgtcaaaaa taaggttatc aagtgagaaa tcaccatgag tgacgactga atccggtgag 2281 aatggcaaaa gtttatgcat ttctttccag acttgttcaa caggccagcc attacgctcg 2341 tcatcaaaat cactcgcatc aaccaaaccg ttattcattc gtgattgcgc ctgagcgaga 2401 cgaaatacgc gatcgctgtt aaaaggacaa ttacaaacag gaatcgaatg caaccggcgc 2461 aggaacactg ccagcgcatc aacaatattt tcacctgaat caggatattc ttctaatacc 2521 tggaatgctg tttttccggg gatcgcagtg gtgagtaacc atgcatcatc aggagtacgg 2581 ataaaatgct tgatggtcgg aagaggcata aattccgtca gccagtttag tctgaccatc 2641 tcatctgtaa catcattggc aacgctacct ttgccatgtt tcagaaacaa ctctggcgca 2701 tcgggcttcc catacaagcg atagattgtc gcacctgatt gcccgacatt atcgcgagcc 2761 catttatacc catataaatc agcatccatg ttggaattta atcgcggcct cgacgtttcc 2821 cgttgaatat ggctcataac accccttgta ttactgttta tgtaagcaga cagttttatt 2881 gttcatgatg atatattttt atcttgtgca atgtaacatc agagattttg agacacgggc 2941 cagagctgc //