LOCUS Exported 3671 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION CT Donor-cassette for SapTrap. ACCESSION . VERSION . KEYWORDS pJW1805 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 3671) 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 3671) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..3671 /organism="synthetic DNA construct" /mol_type="other DNA" primer_bind 51..73 /label=M13/pUC Forward /note="In lacZ gene" primer_bind 65..82 /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind 66..82 /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" primer_bind 89..108 /label=T7 /note="T7 promoter, forward primer" promoter 89..107 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" CDS 216..305 /codon_start=1 /product="3 tandem Myc epitope tags" /label=3xMyc /translation="EQKLISEEDLEQKLISEEDLEQKLISEEDL" promoter complement(410..428) /label=SP6 promoter /note="promoter for bacteriophage SP6 RNA polymerase" primer_bind complement(411..428) /label=SP6 /note="SP6 promoter, forward primer" primer_bind complement(446..462) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(446..462) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(459..481) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 470..486 /label=lac operator /bound_moiety="lac repressor encoded by lacI" /note="The lac repressor binds to the lac operator to inhibit transcription in E. coli. This inhibition can be relieved by adding lactose or isopropyl-beta-D-thiogalactopyranoside (IPTG)." promoter complement(494..524) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 539..560 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(677..694) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(848..1436) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(928..947) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(1574..1948) /codon_start=1 /gene="Sh ble from Streptoalloteichus hindustanus" /product="antibiotic-binding protein" /label=BleoR /note="confers resistance to bleomycin, phleomycin, and Zeocin(TM)" /translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR EFALRDPAGNCVHFVAEEQD" regulatory 1945..1954 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS complement(2154..2948) /codon_start=1 /gene="aph(3')-II (or nptII)" /product="aminoglycoside phosphotransferase from Tn5" /label=NeoR/KanR /note="confers resistance to neomycin, kanamycin, and G418 (Geneticin(R))" /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF" primer_bind complement(2265..2284) /label=Neo-F /note="Neomycin resistance gene, forward primer" primer_bind 2875..2894 /label=Neo-R /note="Neomycin resistance gene, reverse primer" CDS complement(3297..3599) /codon_start=1 /gene="ccdB" /product="CcdB, a bacterial toxin that poisons DNA gyrase" /label=ccdB /note="Plasmids containing the ccdB gene cannot be propagated in standard E. coli strains." /translation="QFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDKV SRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI" primer_bind complement(3399..3418) /label=ccdB-fwd /note="ccdB gene, forward primer" ORIGIN 1 gaaaggggga tgtgctgcaa ggcgattaag ttgggtaacg ccagggtttt cccagtcacg 61 acgttgtaaa acgacggcca gtgaattgta atacgactca ctatagggcg aattgggccc 121 tctagatgca tgctcgagcg gccgccagtg tgatggatat ctgcagaatt cgcccttgaa 181 agctgggtcc tagctcttcg gcgggagccg gatctgaaca aaaacttatt tctgaagagg 241 atcttgagca aaagctcatc tccgaggagg acctcgagca gaagttgatc agcgaggaag 301 acttgatgcg aagagcgacc cagctttctt aagggcgaat tccagcacac tggcggccgt 361 tactagtgga tccgagctcg gtaccaagct tgatgcatag cttgagtatt ctatagtgtc 421 acctaaatag cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc 481 tcacaattcc acacaacata cgagccggaa gcataaagtg taaagcctgg ggtgcctaat 541 gagtgagcta actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc 601 tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg 661 ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag 721 cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag 781 gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc 841 tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc 901 agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc 961 tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt 1021 cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg 1081 ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat 1141 ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag 1201 ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt 1261 ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc 1321 cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta 1381 gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag 1441 atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga 1501 ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa 1561 gttttagcac gtgtcagtcc tgctcctcgg ccacgaagtg cacgcagttg ccggccgggt 1621 cgcgcagggc gaactcccgc ccccacggct gctcgccgat ctcggtcatg gccggcccgg 1681 aggcgtcccg gaagttcgtg gacacgacct ccgaccactc ggcgtacagc tcgtccaggc 1741 cgcgcaccca cacccaggcc agggtgttgt ccggcaccac ctggtcctgg accgcgctga 1801 tgaacagggt cacgtcgtcc cggaccacac cggcgaagtc gtcctccacg aagtcccggg 1861 agaacccgag ccggtcggtc cagaactcga ccgctccggc gacgtcgcgc gcggtgagca 1921 ccggaacggc actggtcaac ttggccatgg tggccctcct cacgtgctat tattgaagca 1981 tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac 2041 aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgatgcggtg tgaaataccg 2101 cacagatgcg taaggagaaa ataccgcatc aggaaattgt aagcgttaat aattcagaag 2161 aactcgtcaa gaaggcgata gaaggcgatg cgctgcgaat cgggagcggc gataccgtaa 2221 agcacgagga agcggtcagc ccattcgccg ccaagctctt cagcaatatc acgggtagcc 2281 aacgctatgt cctgatagcg gtccgccaca cccagccggc cacagtcgat gaatccagaa 2341 aagcggccat tttccaccat gatattcggc aagcaggcat cgccatgggt cacgacgaga 2401 tcctcgccgt cgggcatgct cgccttgagc ctggcgaaca gttcggctgg cgcgagcccc 2461 tgatgctctt cgtccagatc atcctgatcg acaagaccgg cttccatccg agtacgtgct 2521 cgctcgatgc gatgtttcgc ttggtggtcg aatgggcagg tagccggatc aagcgtatgc 2581 agccgccgca ttgcatcagc catgatggat actttctcgg caggagcaag gtgagatgac 2641 aggagatcct gccccggcac ttcgcccaat agcagccagt cccttcccgc ttcagtgaca 2701 acgtcgagca cagctgcgca aggaacgccc gtcgtggcca gccacgatag ccgcgctgcc 2761 tcgtcttgca gttcattcag ggcaccggac aggtcggtct tgacaaaaag aaccgggcgc 2821 ccctgcgctg acagccggaa cacggcggca tcagagcagc cgattgtctg ttgtgcccag 2881 tcatagccga atagcctctc cacccaagcg gccggagaac ctgcgtgcaa tccatcttgt 2941 tcaatcatgc gaaacgatcc tcatcctgtc tcttgatcag agcttgatcc cctgcgccat 3001 cagatccttg gcggcaagaa agccatccag tttactttgc agggcttccc aaccttacca 3061 gagggcgccc cagctggcaa ttccggttcg cttgctgtcc ataaaaccgc ccagtctagc 3121 tatcgccatg taagcccact gcaagctacc tgctttctct ttgcgcttgc gttttccctt 3181 gtccagatag cccagtagct gacattcatc cggggtcagc accgtttctg cggactggct 3241 ttctacgtga aaaggatcta ggtgaagatc ctttttgata atctcatgcc tgacatttat 3301 attccccaga acatcaggtt aatggcgttt ttgatgtcat tttcgcggtg gctgagatca 3361 gccacttctt ccccgataac ggagaccggc acactggcca tatcggtggt catcatgcgc 3421 cagctttcat ccccgatatg caccaccggg taaagttcac gggagacttt atctgacagc 3481 agacgtgcac tggccagggg gatcaccatc cgtcgccccg gcgtgtcaat aatatcactc 3541 tgtacatcca caaacagacg ataacggctc tctcttttat aggtgtaaac cttaaactgc 3601 cgtacgtata ggctgcgcaa ctgttgggaa gggcgatcgg tgcgggcctc ttcgctatta 3661 cgccagctgg c //