LOCUS Exported 2836 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION NT Donor-cassette for SapTrap. ACCESSION . VERSION . KEYWORDS pJW1659 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 2836) 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 2836) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..2836 /organism="synthetic DNA construct" /mol_type="other DNA" primer_bind 91..108 /label=L4440 /note="L4440 vector, forward primer" terminator 267..294 /label=rrnB T2 terminator /note="transcription terminator T2 from the E. coli rrnB gene" terminator 386..472 /gene="Escherichia coli rrnB" /label=rrnB T1 terminator /note="transcription terminator T1 from the E. coli rrnB gene" primer_bind 521..543 /label=M13/pUC Forward /note="In lacZ gene" primer_bind 535..552 /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind 536..552 /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" protein_bind 568..667 /gene="mutant version of attL" /label=attL1 /bound_moiety="LR Clonase(TM)" /note="recombination site for the Gateway(R) LR reaction" CDS 868..891 /codon_start=1 /product="FLAG(R) epitope tag, followed by an enterokinase cleavage site" /label=FLAG /translation="DYKDDDDK" CDS 895..918 /codon_start=1 /product="FLAG(R) epitope tag, followed by an enterokinase cleavage site" /label=FLAG /translation="DYKDDDDK" CDS 922..945 /codon_start=1 /product="FLAG(R) epitope tag, followed by an enterokinase cleavage site" /label=FLAG /translation="DYKDDDDK" protein_bind complement(960..1059) /gene="mutant version of attL" /label=attL2 /bound_moiety="LR Clonase(TM)" /note="recombination site for the Gateway(R) LR reaction" primer_bind complement(1076..1095) /label=T7 /note="T7 promoter, forward primer" promoter complement(1077..1095) /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" primer_bind complement(1100..1116) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(1100..1116) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(1218..1237) /label=pENTR-R /note="pENTR vectors, reverse primer" CDS 1229..2038 /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(1296..1315) /label=Kan-R /note="Kanamycin resistance gene, reverse primer" rep_origin 2185..2773 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 2674..2693 /label=pBR322ori-F /note="pBR322 origin, forward primer" ORIGIN 1 tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgcctttga gtgagctgat 61 accgctcgcc gcagccgaac gaccgagcgc agcgagtcag tgagcgagga agcggaagag 121 cgcccaatac gcaaaccgcc tctccccgcg cgttggccga ttcattaatg cagctggcac 181 gacaggtttc ccgactggaa agcgggcagt gagcgcaacg caattaatac gcgtaccgct 241 agccaggaag agtttgtaga aacgcaaaaa ggccatccgt caggatggcc ttctgcttag 301 tttgatgcct ggcagtttat ggcgggcgtc ctgcccgcca ccctccgggc cgttgcttca 361 caacgttcaa atccgctccc ggcggatttg tcctactcag gagagcgttc accgacaaac 421 aacagataaa acgaaaggcc cagtcttccg actgagcctt tcgttttatt tgatgcctgg 481 cagttcccta ctctcgcgtt aacgctagca tggatgtttt cccagtcacg acgttgtaaa 541 acgacggcca gtcttaagct cgggccccaa ataatgattt tattttgact gatagtgacc 601 tgttcgttgc aacaaattga tgagcaatgc ttttttataa tgccaacttt gtacaaaaaa 661 gcaggctgct cttcgggtgg cggaggaggc agtggtagtg gcagtatgcc taaagatcca 721 gccaaacctc cggccaaggc acaagttgtg ggatggccac cggtgagatc ataccggaag 781 aacgtgatgg tttcctgcca aaaatcaagc ggtggcccgg aggcggcggc gttcgtgaag 841 ggtggtggcg gttctggctc aggatctgat tataaagacg atgacgataa gcgtgactac 901 aaggacgacg acgacaagcg tgattacaag gatgacgatg acaagagaac gcgaagagca 961 cccagctttc ttgtacaaag ttggcattat aagaaagcat tgcttatcaa tttgttgcaa 1021 cgaacaggtc actatcagtc aaaataaaat cattatttgc catccagctg atatccccta 1081 tagtgagtcg tattacatgg tcatagctgt ttcctggcag ctctggcccg tgtctcaaaa 1141 tctctgatgt tacattgcac aagataaaaa tatatcatca tgaacaataa aactgtctgc 1201 ttacataaac agtaatacaa ggggtgttat gagccatatt caacgggaaa cgtcgaggcc 1261 gcgattaaat tccaacatgg atgctgattt atatgggtat aaatgggctc gcgataatgt 1321 cgggcaatca ggtgcgacaa tctatcgctt gtatgggaag cccgatgcgc cagagttgtt 1381 tctgaaacat ggcaaaggta gcgttgccaa tgatgttaca gatgagatgg tcagactaaa 1441 ctggctgacg gaatttatgc ctcttccgac catcaagcat tttatccgta ctcctgatga 1501 tgcatggtta ctcaccactg cgatccccgg aaaaacagca ttccaggtat tagaagaata 1561 tcctgattca ggtgaaaata ttgttgatgc gctggcagtg ttcctgcgcc ggttgcattc 1621 gattcctgtt tgtaattgtc cttttaacag cgatcgcgta tttcgtctcg ctcaggcgca 1681 atcacgaatg aataacggtt tggttgatgc gagtgatttt gatgacgagc gtaatggctg 1741 gcctgttgaa caagtctgga aagaaatgca taaacttttg ccattctcac cggattcagt 1801 cgtcactcat ggtgatttct cacttgataa ccttattttt gacgagggga aattaatagg 1861 ttgtattgat gttggacgag tcggaatcgc agaccgatac caggatcttg ccatcctatg 1921 gaactgcctc ggtgagtttt ctccttcatt acagaaacgg ctttttcaaa aatatggtat 1981 tgataatcct gatatgaata aattgcagtt tcatttgatg ctcgatgagt ttttctaatc 2041 agaattggtt aattggttgt aacactggca gagcattacg ctgacttgac gggacggcgc 2101 aagctcatga ccaaaatccc ttaacgtgag ttacgcgtcg ttccactgag cgtcagaccc 2161 cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt 2221 gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac 2281 tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg ttcttctagt 2341 gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct 2401 gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga 2461 ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac 2521 acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg 2581 agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt 2641 cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc 2701 tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg 2761 gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc 2821 ttttgctcac atgttc //