LOCUS Exported 3898 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION NT Donor-cassette for SapTrap. ACCESSION . VERSION . KEYWORDS pJW1348 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 3898) 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 3898) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..3898 /organism="synthetic DNA construct" /mol_type="other DNA" primer_bind 4..26 /label=M13/pUC Forward /note="In lacZ gene" primer_bind 18..35 /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind 19..35 /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" primer_bind 42..61 /label=T7 /note="T7 promoter, forward primer" promoter 42..60 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" promoter complement(590..608) /label=SP6 promoter /note="promoter for bacteriophage SP6 RNA polymerase" primer_bind complement(591..608) /label=SP6 /note="SP6 promoter, forward primer" primer_bind complement(626..642) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(626..642) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(639..661) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 650..666 /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(674..704) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 719..740 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(857..874) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(1028..1616) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(1108..1127) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(1754..2128) /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 2125..2134 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS complement(2334..3128) /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(2445..2464) /label=Neo-F /note="Neomycin resistance gene, forward primer" primer_bind 3055..3074 /label=Neo-R /note="Neomycin resistance gene, reverse primer" CDS complement(3477..3779) /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(3579..3598) /label=ccdB-fwd /note="ccdB gene, forward primer" ORIGIN 1 tttcccagtc acgacgttgt aaaacgacgg ccagtgaatt gtaatacgac tcactatagg 61 gcgaattggg ccctctagat gcatgctcga gcggccgcca gtgtgatgga tatctgcaga 121 attcgccctt attctgtggc tcttcgggtg gtggcggtgg atcgggagga ggaggttcgg 181 gtggcggagg cagtggaggt ggcggaagtg gcggcggtgg ttcaggagga ggcggatcca 241 cgcgaagagc gtttgatgcc tggcagttta tggcgggatc gaggattacc actacgctac 301 gtgactacgc tgatgtacag atccatactg catgttcatt gacgatctac ctttcctgcg 361 ttatcccctg attctgtggc tcttcgggtg gtggcggtgg atcgggagga ggaggttcgg 421 gtggcggagg cagtggaggt ggcggaagtg gcggcggtgg ttcaggagga ggcggatcca 481 cgcgaagagc gtttgatgaa gggcgaattc aggcctgaat tccagcacac tggcggccgt 541 tactagtgga tccgagctcg gtaccaagct tgatgcatag cttgagtatt ctatagtgtc 601 acctaaatag cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc 661 tcacaattcc acacaacata cgagccggaa gcataaagtg taaagcctgg ggtgcctaat 721 gagtgagcta actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc 781 tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg 841 ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag 901 cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag 961 gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc 1021 tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc 1081 agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc 1141 tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt 1201 cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg 1261 ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat 1321 ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag 1381 ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt 1441 ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc 1501 cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta 1561 gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag 1621 atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga 1681 ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa 1741 gttttagcac gtgtcagtcc tgctcctcgg ccacgaagtg cacgcagttg ccggccgggt 1801 cgcgcagggc gaactcccgc ccccacggct gctcgccgat ctcggtcatg gccggcccgg 1861 aggcgtcccg gaagttcgtg gacacgacct ccgaccactc ggcgtacagc tcgtccaggc 1921 cgcgcaccca cacccaggcc agggtgttgt ccggcaccac ctggtcctgg accgcgctga 1981 tgaacagggt cacgtcgtcc cggaccacac cggcgaagtc gtcctccacg aagtcccggg 2041 agaacccgag ccggtcggtc cagaactcga ccgctccggc gacgtcgcgc gcggtgagca 2101 ccggaacggc actggtcaac ttggccatgg tggccctcct cacgtgctat tattgaagca 2161 tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac 2221 aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgatgcggtg tgaaataccg 2281 cacagatgcg taaggagaaa ataccgcatc aggaaattgt aagcgttaat aattcagaag 2341 aactcgtcaa gaaggcgata gaaggcgatg cgctgcgaat cgggagcggc gataccgtaa 2401 agcacgagga agcggtcagc ccattcgccg ccaagctctt cagcaatatc acgggtagcc 2461 aacgctatgt cctgatagcg gtccgccaca cccagccggc cacagtcgat gaatccagaa 2521 aagcggccat tttccaccat gatattcggc aagcaggcat cgccatgggt cacgacgaga 2581 tcctcgccgt cgggcatgct cgccttgagc ctggcgaaca gttcggctgg cgcgagcccc 2641 tgatgctctt cgtccagatc atcctgatcg acaagaccgg cttccatccg agtacgtgct 2701 cgctcgatgc gatgtttcgc ttggtggtcg aatgggcagg tagccggatc aagcgtatgc 2761 agccgccgca ttgcatcagc catgatggat actttctcgg caggagcaag gtgagatgac 2821 aggagatcct gccccggcac ttcgcccaat agcagccagt cccttcccgc ttcagtgaca 2881 acgtcgagca cagctgcgca aggaacgccc gtcgtggcca gccacgatag ccgcgctgcc 2941 tcgtcttgca gttcattcag ggcaccggac aggtcggtct tgacaaaaag aaccgggcgc 3001 ccctgcgctg acagccggaa cacggcggca tcagagcagc cgattgtctg ttgtgcccag 3061 tcatagccga atagcctctc cacccaagcg gccggagaac ctgcgtgcaa tccatcttgt 3121 tcaatcatgc gaaacgatcc tcatcctgtc tcttgatcag agcttgatcc cctgcgccat 3181 cagatccttg gcggcaagaa agccatccag tttactttgc agggcttccc aaccttacca 3241 gagggcgccc cagctggcaa ttccggttcg cttgctgtcc ataaaaccgc ccagtctagc 3301 tatcgccatg taagcccact gcaagctacc tgctttctct ttgcgcttgc gttttccctt 3361 gtccagatag cccagtagct gacattcatc cggggtcagc accgtttctg cggactggct 3421 ttctacgtga aaaggatcta ggtgaagatc ctttttgata atctcatgcc tgacatttat 3481 attccccaga acatcaggtt aatggcgttt ttgatgtcat tttcgcggtg gctgagatca 3541 gccacttctt ccccgataac ggagaccggc acactggcca tatcggtggt catcatgcgc 3601 cagctttcat ccccgatatg caccaccggg taaagttcac gggagacttt atctgacagc 3661 agacgtgcac tggccagggg gatcaccatc cgtcgccccg gcgtgtcaat aatatcactc 3721 tgtacatcca caaacagacg ataacggctc tctcttttat aggtgtaaac cttaaactgc 3781 cgtacgtata ggctgcgcaa ctgttgggaa gggcgatcgg tgcgggcctc ttcgctatta 3841 cgccagctgg cgaaaggggg atgtgctgca aggcgattaa gttgggtaac gccagggt //