LOCUS Exported File 7685 bp ds-DNA circular SYN 08-FEB-2012 DEFINITION Addgene ID 34978. ACCESSION . VERSION . KEYWORDS pDS191 SOURCE synthetic DNA construct ORGANISM recombinant plasmid REFERENCE 1 (bases 1 to 7685) AUTHORS Devin Strickland TITLE Direct Submission JOURNAL Exported Feb 8, 2012 from SnapGene 0.6.2.2 http://www.snapgene.com COMMENT TULIPs cassette inserted into pGREG503 between SacI and XhoI FEATURES Location/Qualifiers source 1..7685 /organism="recombinant plasmid" /lab_host="Saccharomyces cerevisiae" /mol_type="other DNA" promoter 1..403 /gene="TEF Promoter" /note="TEF prom" /note="color: #c0c0c0; direction: RIGHT" CDS 473..1054 /codon_start=1 /note="ePDZb" /note="color: #666699" /translation="MPELGFSISGGVGGRGNPFRPDDDGIFVTRVQPEGPASKLLQPGD KIIQANGYSFINIEHGQAVSLLKTFQNTVELIIVREVGNGAKQEIRVRVEKDGGSGGVS SVPTNLEVVAATPTSLLISWDAYYDSHVSYYRITYGETGGNSPVQEFTVPGSKSTATIS GLKPGVDYTITVYAHYNYHYYSSPISINYRT" terminator 1067..1314 /note="CYC1 term" /note="color: #c0c0c0" protein_bind 1327..1360 /bound_moiety="Cre recombinase" /note="loxP" /note="Cre-mediated recombination occurs in the 8-bp core sequence (GCATACAT)." /note="color: #31849b; direction: RIGHT" CDS complement(1621..2430) /codon_start=1 /gene="aph(3')-Ia" /product="aminoglycoside phosphotransferase" /note="KanMX" /note="confers resistance to kanamycin" /note="color: #ccffcc" /translation="MGKEKTHVSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF" protein_bind 2829..2862 /bound_moiety="Cre recombinase" /note="loxP" /note="Cre-mediated recombination occurs in the 8-bp core sequence (GCATACAT)." /note="color: #31849b; direction: RIGHT" primer_bind complement(2927..2943) /note="M13 fwd" /note="common sequencing primer, one of multiple similar variants" /note="color: #a020f0; direction: LEFT" rep_origin 3088..3543 /direction=RIGHT /note="f1 ori" /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" /note="color: #ffff00" CDS complement(3674..4477) /codon_start=1 /gene="S. cerevisiae URA3" /product="orotidine-5'-phosphate decarboxylase, required for uracil biosynthesis" /note="URA3" /note="yeast auxotrophic marker, counterselectable with 5-fluoroorotic acid (5-FOA)" /note="color: #ff7f50" /translation="MSKATYKERAATHPSPVAAKLFNIMHEKQTNLCASLDVRTTKELL ELVEALGPKICLLKTHVDILTDFSMEGTVKPLKALSAKYNFLLFEDRKFADIGNTVKLQ YSAGVYRIAEWADITNAHGVVGPGIVSGLKQAAEEVTKEPRGLLMLAELSCKGSLSTGE YTKGTVDIAKSDKDFVIGFIAQRDMGGRDEGYDWLIMTPGVGLDDKGDALGQQYRTVDD VVSTGSDIIIVGRGLFAKGRDAKVEGERYRKAGWEAYLRRCGQQN" promoter complement(4478..4698) /gene="S. cerevisiae URA3" /note="URA3 promoter" /note="color: #ffffff; direction: LEFT" misc_feature 4963..5466 /note="CEN/ARS" /note="S. cerevisiae CEN6 centromere fused to an autonomously replicating sequence" /note="color: #454f59" promoter 5503..5607 /gene="bla" /note="AmpR promoter" /note="color: #ffffff; direction: RIGHT" CDS 5608..6468 /codon_start=1 /gene="bla" /product="beta-lactamase" /note="AmpR" /note="confers resistance to ampicillin, carbenicillin, and related antibiotics" /note="This feature has 2 segments: 1: 5608 .. 5676 / #ccffcc / signal sequence 2: 5677 .. 6468 / #ccffcc Cleavage site after base 5676" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS LIKHW" rep_origin 6639..7227 /direction=RIGHT /note="ori" /note="high-copy-number colE1/pMB1/pBR322/pUC origin of replication" /note="color: #ffff00" primer_bind 7613..7629 /note="M13 rev" /note="common sequencing primer, one of multiple similar variants" /note="color: #a020f0; direction: RIGHT" ORIGIN 1 catagcttca aaatgtttct actccttttt tactcttcca gattttctcg gactccgcgc 61 atcgccgtac cacttcaaaa cacccaagca cagcatacta aatttcccct ctttcttcct 121 ctagggtgtc gttaattacc cgtactaaag gtttggaaaa gaaaaaagag accgcctcgt 181 ttctttttct tcgtcgaaaa aggcaataaa aatttttatc acgtttcttt ttcttgaaaa 241 tttttttttt gatttttttc tctttcgatg acctcccatt gatatttaag ttaataaacg 301 gtcttcaatt tctcaagttt cagtttcatt tttcttgttc tattacaact ttttttactt 361 cttgctcatt agaaagaaag catagcaatc taatctaagt tttctagaac tagtggatcc 421 cccgggctgc aggaattgga tatcaagctt atcgataccg tcgacctcga caatgccaga 481 acttggattt agcatatcag gtggtgtcgg gggtagagga aacccattca gacctgatga 541 tgatggtata tttgtaacaa gggtacaacc tgaaggacca gcatcaaaat tactgcagcc 601 aggtgataaa attattcagg ctaatggcta cagttttata aatattgaac atggacaagc 661 agtgtccttg ctaaaaactt tccagaatac agttgaactc atcattgtac gagaagttgg 721 taacggtgca aaacaagaga ttcgagtgag ggttgaaaaa gacggtggtt ctggaggcgt 781 ttcttctgtt ccgaccaatc tggaagttgt tgctgcgacc ccgactagcc tgctgatcag 841 ctgggatgcg tactacgact ctcatgtttc ttattaccgt atcacgtacg gtgaaaccgg 901 tggtaactcc ccggttcagg aattcactgt acctggttcc aagtctactg ctaccatcag 961 cggcctgaaa ccgggtgttg actataccat cactgtatac gcacattaca actaccatta 1021 ctactctagc ccaatctcga ttaactaccg tacctctaga ctcgagtcat gtaattagtt 1081 atgtcacgct tacattcacg ccctcccccc acatccgctc taaccgaaaa ggaaggagtt 1141 agacaacctg aagtctaggt ccctatttat ttttttatag ttatgttagt attaagaacg 1201 ttatttatat ttcaaatttt tctttttttt ctgtacagac gcgtgtacgc atgtaacatt 1261 atactgaaaa ccttgcttga gaaggttttg ggacgctcga aggctttaat ttgcggccgg 1321 tacctaataa cttcgtatag catacattat acgaagttat attaagggtt ctcgacgttt 1381 tcgacactgg atggcggcgt tagtatcgaa tcgacagcag tatagcgacc agcattcaca 1441 tacgattgac gcatgatatt actttctgcg cacttaactt cgcatctggg cagatgatgt 1501 cgaggcgaaa aaaaatataa atcacgctaa catttgatta aaatagaaca actacaatat 1561 aaaaaaacta tacaaatgac aagttcttga aaacaagaat ctttttattg tcagtactga 1621 ttagaaaaac tcatcgagca tcaaatgaaa ctgcaattta ttcatatcag gattatcaat 1681 accatatttt tgaaaaagcc gtttctgtaa tgaaggagaa aactcaccga ggcagttcca 1741 taggatggca agatcctggt atcggtctgc gattccgact cgtccaacat caatacaacc 1801 tattaatttc ccctcgtcaa aaataaggtt atcaagtgag aaatcaccat gagtgacgac 1861 tgaatccggt gagaatggca aaagcttatg catttctttc cagacttgtt caacaggcca 1921 gccattacgc tcgtcatcaa aatcactcgc atcaaccaaa ccgttattca ttcgtgattg 1981 cgcctgagcg agacgaaata cgcgatcgct gttaaaagga caattacaaa caggaatcga 2041 atgcaaccgg cgcaggaaca ctgccagcgc atcaacaata ttttcacctg aatcaggata 2101 ttcttctaat acctggaatg ctgttttgcc ggggatcgca gtggtgagta accatgcatc 2161 atcaggagta cggataaaat gcttgatggt cggaagaggc ataaattccg tcagccagtt 2221 tagtctgacc atctcatctg taacatcatt ggcaacgcta cctttgccat gtttcagaaa 2281 caactctggc gcatcgggct tcccatacaa tcgatagatt gtcgcacctg attgcccgac 2341 attatcgcga gcccatttat acccatataa atcagcatcc atgttggaat ttaatcgcgg 2401 cctcgaaacg tgagtctttt ccttacccat ggttgtttat gttcggatgt gatgtgagaa 2461 ctgtatccta gcaagatttt aaaaggaagt atatgaaaga agaacctcag tggcaaatcc 2521 taacctttta tatttctcta caggggcgcg gcgtggggac aattcaacgc gtctgtgagg 2581 ggagcgtttc cctgctcgca ggtctgcagc gaggagccgt aatttttgct tcgcgccgtg 2641 cggccatcaa aatgtatgga tgcaaatgat tatacatggg gatgtatggg ctaaatgtac 2701 gggcgacagt cacatcatgc ccctgagctg cgcacgtcaa gactgtcaag gagggtattc 2761 tgggcctcca tgtcgctggc cgggtgaccc ggcggggacg aggcaagcta aacagatctc 2821 tagacctaat aacttcgtat agcatacatt atacgaagtt atattaaggg ttgtcttaat 2881 taagggtacc caattcgccc tatagtgagt cgtattacgc gcgctcactg gccgtcgttt 2941 tacaacgtcg tgactgggaa aaccctggcg ttacccaact taatcgcctt gcagcacatc 3001 cccctttcgc cagctggcgt aatagcgaag aggcccgcac cgatcgccct tcccaacagt 3061 tgcgcagcct gaatggcgaa tggcgcgacg cgccctgtag cggcgcatta agcgcggcgg 3121 gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccctagcg cccgctcctt 3181 tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc 3241 gggggctccc tttagggttc cgatttagtg ctttacggca cctcgacccc aaaaaacttg 3301 attagggtga tggttcacgt agtgggccat cgccctgata gacggttttt cgccctttga 3361 cgttggagtc cacgttcttt aatagtggac tcttgttcca aactggaaca acactcaacc 3421 ctatctcggt ctattctttt gatttataag ggattttgcc gatttcggcc tattggttaa 3481 aaaatgagct gatttaacaa aaatttaacg cgaattttaa caaaatatta acgtttacaa 3541 tttcctgatg cggtattttc tccttacgca tctgtgcggt atttcacacc gcatagggta 3601 ataactgata taattaaatt gaagctctaa tttgtgagtt tagtatacat gcatttactt 3661 ataatacagt tttttagttt tgctggccgc atcttctcaa atatgcttcc cagcctgctt 3721 ttctgtaacg ttcaccctct accttagcat cccttccctt tgcaaatagt cctcttccaa 3781 caataataat gtcagatcct gtagagacca catcatccac ggttctatac tgttgaccca 3841 atgcgtctcc cttgtcatct aaacccacac cgggtgtcat aatcaaccaa tcgtaacctt 3901 catctcttcc acccatgtct ctttgagcaa taaagccgat aacaaaatct ttgtcgctct 3961 tcgcaatgtc aacagtaccc ttagtatatt ctccagtaga tagggagccc ttgcatgaca 4021 attctgctaa catcaaaagg cctctaggtt cctttgttac ttcttctgcc gcctgcttca 4081 aaccgctaac aatacctggg cccaccacac cgtgtgcatt cgtaatgtct gcccattctg 4141 ctattctgta tacacccgca gagtactgca atttgactgt attaccaatg tcagcaaatt 4201 ttctgtcttc gaagagtaaa aaattgtact tggcggataa tgcctttagc ggcttaactg 4261 tgccctccat ggaaaaatca gtcaagatat ccacatgtgt ttttagtaaa caaattttgg 4321 gacctaatgc ttcaactaac tccagtaatt ccttggtggt acgaacatcc aatgaagcac 4381 acaagtttgt ttgcttttcg tgcatgatat taaatagctt ggcagcaaca ggactaggat 4441 gagtagcagc acgttcctta tatgtagctt tcgacatgat ttatcttcgt ttcctgcagg 4501 tttttgttct gtgcagttgg gttaagaata ctgggcaatt tcatgtttct tcaacactac 4561 atatgcgtat atataccaat ctaagtctgt gctccttcct tcgttcttcc ttctgttcgg 4621 agattaccga atcaaaaaaa tttcaaagaa accgaaatca aaaaaaagaa taaaaaaaaa 4681 atgatgaatt gaattgaaaa gctgtggtat ggtgcactct cagtacaatc tgctctgatg 4741 ccgcatagtt aagccagccc cgacacccgc caacacccgc tgacgcgccc tgacgggctt 4801 gtctgctccc ggcatccgct tacagacaag ctgtgaccgt ctccgggagc tgcatgtgtc 4861 agaggttttc accgtcatca ccgaaacgcg cgagacgaaa gggcctcgtg atacgcctat 4921 ttttataggt taatgtcatg ataataatgg tttcttagga cggatcgctt gcctgtaact 4981 tacacgcgcc tcgtatcttt taatgatgga ataatttggg aatttactct gtgtttattt 5041 atttttatgt tttgtatttg gattttagaa agtaaataaa gaaggtagaa gagttacgga 5101 atgaagaaaa aaaaataaac aaaggtttaa aaaatttcaa caaaaagcgt actttacata 5161 tatatttatt agacaagaaa agcagattaa atagatatac attcgattaa cgataagtaa 5221 aatgtaaaat cacaggattt tcgtgtgtgg tcttctacac agacaagatg aaacaattcg 5281 gcattaatac ctgagagcag gaagagcaag ataaaaggta gtatttgttg gcgatccccc 5341 tagagtcttt tacatcttcg gaaaacaaaa actatttttt ctttaatttc tttttttact 5401 ttctattttt aatttatata tttatattaa aaaatttaaa ttataattat ttttatagca 5461 cgtgatgaaa aggacccagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt 5521 ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg 5581 cttcaataat attgaaaaag gaagagtatg agtattcaac atttccgtgt cgcccttatt 5641 cccttttttg cggcattttg ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta 5701 aaagatgctg aagatcagtt gggtgcacga gtgggttaca tcgaactgga tctcaacagc 5761 ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc caatgatgag cacttttaaa 5821 gttctgctat gtggcgcggt attatcccgt attgacgccg ggcaagagca actcggtcgc 5881 cgcatacact attctcagaa tgacttggtt gagtactcac cagtcacaga aaagcatctt 5941 acggatggca tgacagtaag agaattatgc agtgctgcca taaccatgag tgataacact 6001 gcggccaact tacttctgac aacgatcgga ggaccgaagg agctaaccgc ttttttgcac 6061 aacatggggg atcatgtaac tcgccttgat cgttgggaac cggagctgaa tgaagccata 6121 ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta 6181 ttaactggcg aactacttac tctagcttcc cggcaacaat taatagactg gatggaggcg 6241 gataaagttg caggaccact tctgcgctcg gcccttccgg ctggctggtt tattgctgat 6301 aaatctggag ccggtgagcg tgggtctcgc ggtatcattg cagcactggg gccagatggt 6361 aagccctccc gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga 6421 aatagacaga tcgctgagat aggtgcctca ctgattaagc attggtaact gtcagaccaa 6481 gtttactcat atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag 6541 gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac 6601 tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc 6661 gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat 6721 caagagctac caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat 6781 actgtccttc tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct 6841 acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt 6901 cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg 6961 gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta 7021 cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg 7081 gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg 7141 tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc 7201 tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg 7261 gccttttgct ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat 7321 aaccgtatta ccgcctttga gtgagctgat accgctcgcc gcagccgaac gaccgagcgc 7381 agcgagtcag tgagcgagga agcggaagag cgcccaatac gcaaaccgcc tctccccgcg 7441 cgttggccga ttcattaatg cagctggcac gacaggtttc ccgactggaa agcgggcagt 7501 gagcgcaacg caattaatgt gagttacctc actcattagg caccccaggc tttacacttt 7561 atgcttccgg ctcctatgtt gtgtggaatt gtgagcggat aacaatttca cacaggaaac 7621 agctatgacc atgattacgc caagcgcgca attaaccctc actaaaggga acaaaagctg 7681 gagct //