LOCUS Exported File 8752 bp ds-DNA circular SYN 08-FEB-2012 DEFINITION Addgene ID 34977. ACCESSION . VERSION . KEYWORDS pDS190 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 8752) 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..8752 /organism="synthetic DNA construct" /lab_host="Saccharomyces cerevisiae" /mol_type="other DNA" promoter 1..1463 /gene=" " /note="ADH prom" /note="color: #c0c0c0; direction: RIGHT" CDS 1540..2121 /codon_start=1 /note="ePDZb" /note="color: #666699" /translation="MPELGFSISGGVGGRGNPFRPDDDGIFVTRVQPEGPASKLLQPGD KIIQANGYSFINIEHGQAVSLLKTFQNTVELIIVREVGNGAKQEIRVRVEKDGGSGGVS SVPTNLEVVAATPTSLLISWDAYYDSHVSYYRITYGETGGNSPVQEFTVPGSKSTATIS GLKPGVDYTITVYAHYNYHYYSSPISINYRT" terminator 2134..2381 /note="CYC1 term" /note="color: #c0c0c0" protein_bind 2394..2427 /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(2688..3497) /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 3896..3929 /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(3994..4010) /note="M13 fwd" /note="common sequencing primer, one of multiple similar variants" /note="color: #a020f0; direction: LEFT" rep_origin 4155..4610 /direction=RIGHT /note="f1 ori" /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" /note="color: #ffff00" CDS complement(4741..5544) /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(5545..5765) /gene="S. cerevisiae URA3" /note="URA3 promoter" /note="color: #ffffff; direction: LEFT" misc_feature 6030..6533 /note="CEN/ARS" /note="S. cerevisiae CEN6 centromere fused to an autonomously replicating sequence" /note="color: #454f59" promoter 6570..6674 /gene="bla" /note="AmpR promoter" /note="color: #ffffff; direction: RIGHT" CDS 6675..7535 /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: 6675 .. 6743 / #ccffcc / signal sequence 2: 6744 .. 7535 / #ccffcc Cleavage site after base 6743" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS LIKHW" rep_origin 7706..8294 /direction=RIGHT /note="ori" /note="high-copy-number colE1/pMB1/pBR322/pUC origin of replication" /note="color: #ffff00" primer_bind 8680..8696 /note="M13 rev" /note="common sequencing primer, one of multiple similar variants" /note="color: #a020f0; direction: RIGHT" ORIGIN 1 cgccgggatc gaagaaatga tggtaaatga aataggaaat caaggagcat gaaggcaaaa 61 gacaaatata agggtcgaac gaaaaataaa gtgaaaagtg ttgatatgat gtatttggct 121 ttgcggcgcc gaaaaaacga gtttacgcaa ttgcacaatc atgctgactc tgtggcggac 181 ccgcgctctt gccggcccgg cgataacgct gggcgtgagg ctgtgcccgg cggagttttt 241 tgcgcctgca ttttccaagg tttaccctgc gctaaggggc gagattggag aagcaataag 301 aatgccggtt ggggttgcga tgatgacgac cacgacaact ggtgtcatta tttaagttgc 361 cgaaagaacc tgagtgcatt tgcaacatga gtatactaga agaatgagcc aagacttgcg 421 agacgcgagt ttgccggtgg tgcgaacaat agagcgacca tgaccttgaa ggtgagacgc 481 gcataaccgc tagagtactt tgaagaggaa acagcaatag ggttgctacc agtataaata 541 gacaggtaca tacaacactg gaaatggttg tctgtttgag tacgctttca attcatttgg 601 gtgtgcactt tattatgtta caatatggaa gggaacttta cacttctcct atgcacatat 661 attaattaaa gtccaatgct agtagagaag gggggtaaca cccctccgcg ctcttttccg 721 atttttttct aaaccgtgga atatttcgga tatccttttg ttgtttccgg gtgtacaata 781 tggacttcct cttttctggc aaccaaaccc atacatcggg attcctataa taccttcgtt 841 ggtctcccta acatgtaggt ggcggagggg agatatacaa tagaacagat accagacaag 901 acataatggg ctaaacaaga ctacaccaat tacactgcct cattgatggt ggtacataac 961 gaactaatac tgtagcccta gacttgatag ccatcatcat atcgaagttt cactaccctt 1021 tttccatttg ccatctattg aagtaataat aggcgcatgc aacttctttt cttttttttt 1081 cttttctctc tcccccgttg ttgtctcacc atatccgcaa tgacaaaaaa atgatggaag 1141 acactaaagg aaaaaattaa cgacaaagac agcaccaaca gatgtcgttg ttccagagct 1201 gatgaggggt atctcgaagc acacgaaact ttttccttcc ttcattcacg cacactactc 1261 tctaatgagc aacggtatac ggccttcctt ccagttactt gaatttgaaa taaaaaaaag 1321 tttgctgtct tgctatcaag tataaataga cctgcaatta ttaatctttt gtttcctcgt 1381 cattgttctc gttccctttc ttccttgttt ctttttctgc acaatatttc aagctatacc 1441 aagcatacaa tcaactccaa gctggccgct ctagaactag tggatccccc gggctgcagg 1501 aattggatat caagcttatc gataccgtcg acctcgacaa tgccagaact tggatttagc 1561 atatcaggtg gtgtcggggg tagaggaaac ccattcagac ctgatgatga tggtatattt 1621 gtaacaaggg tacaacctga aggaccagca tcaaaattac tgcagccagg tgataaaatt 1681 attcaggcta atggctacag ttttataaat attgaacatg gacaagcagt gtccttgcta 1741 aaaactttcc agaatacagt tgaactcatc attgtacgag aagttggtaa cggtgcaaaa 1801 caagagattc gagtgagggt tgaaaaagac ggtggttctg gaggcgtttc ttctgttccg 1861 accaatctgg aagttgttgc tgcgaccccg actagcctgc tgatcagctg ggatgcgtac 1921 tacgactctc atgtttctta ttaccgtatc acgtacggtg aaaccggtgg taactccccg 1981 gttcaggaat tcactgtacc tggttccaag tctactgcta ccatcagcgg cctgaaaccg 2041 ggtgttgact ataccatcac tgtatacgca cattacaact accattacta ctctagccca 2101 atctcgatta actaccgtac ctctagactc gagtcatgta attagttatg tcacgcttac 2161 attcacgccc tccccccaca tccgctctaa ccgaaaagga aggagttaga caacctgaag 2221 tctaggtccc tatttatttt tttatagtta tgttagtatt aagaacgtta tttatatttc 2281 aaatttttct tttttttctg tacagacgcg tgtacgcatg taacattata ctgaaaacct 2341 tgcttgagaa ggttttggga cgctcgaagg ctttaatttg cggccggtac ctaataactt 2401 cgtatagcat acattatacg aagttatatt aagggttctc gacgttttcg acactggatg 2461 gcggcgttag tatcgaatcg acagcagtat agcgaccagc attcacatac gattgacgca 2521 tgatattact ttctgcgcac ttaacttcgc atctgggcag atgatgtcga ggcgaaaaaa 2581 aatataaatc acgctaacat ttgattaaaa tagaacaact acaatataaa aaaactatac 2641 aaatgacaag ttcttgaaaa caagaatctt tttattgtca gtactgatta gaaaaactca 2701 tcgagcatca aatgaaactg caatttattc atatcaggat tatcaatacc atatttttga 2761 aaaagccgtt tctgtaatga aggagaaaac tcaccgaggc agttccatag gatggcaaga 2821 tcctggtatc ggtctgcgat tccgactcgt ccaacatcaa tacaacctat taatttcccc 2881 tcgtcaaaaa taaggttatc aagtgagaaa tcaccatgag tgacgactga atccggtgag 2941 aatggcaaaa gcttatgcat ttctttccag acttgttcaa caggccagcc attacgctcg 3001 tcatcaaaat cactcgcatc aaccaaaccg ttattcattc gtgattgcgc ctgagcgaga 3061 cgaaatacgc gatcgctgtt aaaaggacaa ttacaaacag gaatcgaatg caaccggcgc 3121 aggaacactg ccagcgcatc aacaatattt tcacctgaat caggatattc ttctaatacc 3181 tggaatgctg ttttgccggg gatcgcagtg gtgagtaacc atgcatcatc aggagtacgg 3241 ataaaatgct tgatggtcgg aagaggcata aattccgtca gccagtttag tctgaccatc 3301 tcatctgtaa catcattggc aacgctacct ttgccatgtt tcagaaacaa ctctggcgca 3361 tcgggcttcc catacaatcg atagattgtc gcacctgatt gcccgacatt atcgcgagcc 3421 catttatacc catataaatc agcatccatg ttggaattta atcgcggcct cgaaacgtga 3481 gtcttttcct tacccatggt tgtttatgtt cggatgtgat gtgagaactg tatcctagca 3541 agattttaaa aggaagtata tgaaagaaga acctcagtgg caaatcctaa ccttttatat 3601 ttctctacag gggcgcggcg tggggacaat tcaacgcgtc tgtgagggga gcgtttccct 3661 gctcgcaggt ctgcagcgag gagccgtaat ttttgcttcg cgccgtgcgg ccatcaaaat 3721 gtatggatgc aaatgattat acatggggat gtatgggcta aatgtacggg cgacagtcac 3781 atcatgcccc tgagctgcgc acgtcaagac tgtcaaggag ggtattctgg gcctccatgt 3841 cgctggccgg gtgacccggc ggggacgagg caagctaaac agatctctag acctaataac 3901 ttcgtatagc atacattata cgaagttata ttaagggttg tcttaattaa gggtacccaa 3961 ttcgccctat agtgagtcgt attacgcgcg ctcactggcc gtcgttttac aacgtcgtga 4021 ctgggaaaac cctggcgtta cccaacttaa tcgccttgca gcacatcccc ctttcgccag 4081 ctggcgtaat agcgaagagg cccgcaccga tcgcccttcc caacagttgc gcagcctgaa 4141 tggcgaatgg cgcgacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac 4201 gcgcagcgtg accgctacac ttgccagcgc cctagcgccc gctcctttcg ctttcttccc 4261 ttcctttctc gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt 4321 agggttccga tttagtgctt tacggcacct cgaccccaaa aaacttgatt agggtgatgg 4381 ttcacgtagt gggccatcgc cctgatagac ggtttttcgc cctttgacgt tggagtccac 4441 gttctttaat agtggactct tgttccaaac tggaacaaca ctcaacccta tctcggtcta 4501 ttcttttgat ttataaggga ttttgccgat ttcggcctat tggttaaaaa atgagctgat 4561 ttaacaaaaa tttaacgcga attttaacaa aatattaacg tttacaattt cctgatgcgg 4621 tattttctcc ttacgcatct gtgcggtatt tcacaccgca tagggtaata actgatataa 4681 ttaaattgaa gctctaattt gtgagtttag tatacatgca tttacttata atacagtttt 4741 ttagttttgc tggccgcatc ttctcaaata tgcttcccag cctgcttttc tgtaacgttc 4801 accctctacc ttagcatccc ttccctttgc aaatagtcct cttccaacaa taataatgtc 4861 agatcctgta gagaccacat catccacggt tctatactgt tgacccaatg cgtctccctt 4921 gtcatctaaa cccacaccgg gtgtcataat caaccaatcg taaccttcat ctcttccacc 4981 catgtctctt tgagcaataa agccgataac aaaatctttg tcgctcttcg caatgtcaac 5041 agtaccctta gtatattctc cagtagatag ggagcccttg catgacaatt ctgctaacat 5101 caaaaggcct ctaggttcct ttgttacttc ttctgccgcc tgcttcaaac cgctaacaat 5161 acctgggccc accacaccgt gtgcattcgt aatgtctgcc cattctgcta ttctgtatac 5221 acccgcagag tactgcaatt tgactgtatt accaatgtca gcaaattttc tgtcttcgaa 5281 gagtaaaaaa ttgtacttgg cggataatgc ctttagcggc ttaactgtgc cctccatgga 5341 aaaatcagtc aagatatcca catgtgtttt tagtaaacaa attttgggac ctaatgcttc 5401 aactaactcc agtaattcct tggtggtacg aacatccaat gaagcacaca agtttgtttg 5461 cttttcgtgc atgatattaa atagcttggc agcaacagga ctaggatgag tagcagcacg 5521 ttccttatat gtagctttcg acatgattta tcttcgtttc ctgcaggttt ttgttctgtg 5581 cagttgggtt aagaatactg ggcaatttca tgtttcttca acactacata tgcgtatata 5641 taccaatcta agtctgtgct ccttccttcg ttcttccttc tgttcggaga ttaccgaatc 5701 aaaaaaattt caaagaaacc gaaatcaaaa aaaagaataa aaaaaaaatg atgaattgaa 5761 ttgaaaagct gtggtatggt gcactctcag tacaatctgc tctgatgccg catagttaag 5821 ccagccccga cacccgccaa cacccgctga cgcgccctga cgggcttgtc tgctcccggc 5881 atccgcttac agacaagctg tgaccgtctc cgggagctgc atgtgtcaga ggttttcacc 5941 gtcatcaccg aaacgcgcga gacgaaaggg cctcgtgata cgcctatttt tataggttaa 6001 tgtcatgata ataatggttt cttaggacgg atcgcttgcc tgtaacttac acgcgcctcg 6061 tatcttttaa tgatggaata atttgggaat ttactctgtg tttatttatt tttatgtttt 6121 gtatttggat tttagaaagt aaataaagaa ggtagaagag ttacggaatg aagaaaaaaa 6181 aataaacaaa ggtttaaaaa atttcaacaa aaagcgtact ttacatatat atttattaga 6241 caagaaaagc agattaaata gatatacatt cgattaacga taagtaaaat gtaaaatcac 6301 aggattttcg tgtgtggtct tctacacaga caagatgaaa caattcggca ttaatacctg 6361 agagcaggaa gagcaagata aaaggtagta tttgttggcg atccccctag agtcttttac 6421 atcttcggaa aacaaaaact attttttctt taatttcttt ttttactttc tatttttaat 6481 ttatatattt atattaaaaa atttaaatta taattatttt tatagcacgt gatgaaaagg 6541 acccaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa 6601 tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt 6661 gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg 6721 cattttgcct tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctgaag 6781 atcagttggg tgcacgagtg ggttacatcg aactggatct caacagcggt aagatccttg 6841 agagttttcg ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg 6901 gcgcggtatt atcccgtatt gacgccgggc aagagcaact cggtcgccgc atacactatt 6961 ctcagaatga cttggttgag tactcaccag tcacagaaaa gcatcttacg gatggcatga 7021 cagtaagaga attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac 7081 ttctgacaac gatcggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc 7141 atgtaactcg ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc 7201 gtgacaccac gatgcctgta gcaatggcaa caacgttgcg caaactatta actggcgaac 7261 tacttactct agcttcccgg caacaattaa tagactggat ggaggcggat aaagttgcag 7321 gaccacttct gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg 7381 gtgagcgtgg gtctcgcggt atcattgcag cactggggcc agatggtaag ccctcccgta 7441 tcgtagttat ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg 7501 ctgagatagg tgcctcactg attaagcatt ggtaactgtc agaccaagtt tactcatata 7561 tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt 7621 ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc 7681 ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct 7741 tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa 7801 ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact gtccttctag 7861 tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc 7921 tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg 7981 actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca 8041 cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat 8101 gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg 8161 tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc 8221 ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc 8281 ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc 8341 cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg 8401 cctttgagtg agctgatacc gctcgccgca gccgaacgac cgagcgcagc gagtcagtga 8461 gcgaggaagc ggaagagcgc ccaatacgca aaccgcctct ccccgcgcgt tggccgattc 8521 attaatgcag ctggcacgac aggtttcccg actggaaagc gggcagtgag cgcaacgcaa 8581 ttaatgtgag ttacctcact cattaggcac cccaggcttt acactttatg cttccggctc 8641 ctatgttgtg tggaattgtg agcggataac aatttcacac aggaaacagc tatgaccatg 8701 attacgccaa gcgcgcaatt aaccctcact aaagggaaca aaagctggag ct //