LOCUS Exported File 5027 bp ds-DNA circular SYN 29-FEB-2012 DEFINITION Addgene ID 34984. ACCESSION . VERSION . KEYWORDS pDS295 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 5027) AUTHORS Glotzer Lab TITLE Direct Submission JOURNAL Exported Feb 29, 2012 from SnapGene 0.7.0 http://www.snapgene.com COMMENT TULIPs cassette cloned into YIplac211 between KpnI and SacI FEATURES Location/Qualifiers source 1..5027 /organism="synthetic DNA construct" /lab_host="Saccharomyces cerevisiae" /mol_type="other DNA" promoter 22..471 /gene=" " /note="GAL1 prom" /note="color: #c0c0c0; direction: RIGHT" CDS 556..1137 /codon_start=1 /note="ePDZb" /note="color: #666699" /translation="MPELGFSISGGVGGRGNPFRPDDDGIFVTRVQPEGPASKLLQPGD KIIQANGYSFINIEHGQAVSLLKTFQNTVELIIVREVGNGAKQEIRVRVEKDGGSGGVS SVPTNLEVVAATPTSLLISWDAYYDSHVSYYRITYGETGGNSPVQEFTVPGSKSTATIS GLKPGVDYTITVYAHYNYHYYSSPISINYRT" terminator 1150..1397 /note="CYC1 term" /note="color: #ffff99" CDS complement(1731..2534) /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(2535..2755) /gene="S. cerevisiae URA3" /note="URA3 promoter" /note="color: #ffffff; direction: LEFT" promoter 2848..2952 /gene="bla" /note="AmpR promoter" /note="color: #ffffff; direction: RIGHT" CDS 2953..3813 /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: 2953 .. 3021 / #ccffcc / signal sequence 2: 3022 .. 3813 / #ccffcc Cleavage site after base 3021" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS LIKHW" rep_origin 3984..4572 /direction=RIGHT /note="ori" /note="high-copy-number colE1/pMB1/pBR322/pUC origin of replication" /note="color: #ffff00" primer_bind 4958..4974 /note="M13 rev" /note="common sequencing primer, one of multiple similar variants" /note="color: #a020f0; direction: RIGHT" ORIGIN 1 cgtttaaacg gcgcgcctag tacggattag aagccgccga gcgggtgaca gccctccgaa 61 ggaagactct cctccgtgcg tcctcgtctt caccggtcgc gttcctgaaa cgcagatgtg 121 cctcgcgccg cactgctccg aacaataaag attctacaat actagctttt atggttatga 181 agaggaaaaa ttggcagtaa cctggcccca caaaccttca aatgaacgaa tcaaattaac 241 aaccatagga tgataatgcg attagttttt tagccttatt tctggggtaa ttaatcagcg 301 aagcgatgat ttttgatcta ttaacagata tataaatgca aaaactgcat aaccacttta 361 actaatactt tcaacatttt cggtttgtat tacttcttat tcaaatgtaa taaaagtatc 421 aacaaaaaat tgttaatata cctctatact ttaacgtcaa ggagaaaaac cccggattct 481 gaagcggccg cactagtgga tcccccgggc tgcaggaatt ggatatcaag cttatcgata 541 ccgtcgacct cgacaatgcc agaacttgga tttagcatat caggtggtgt cgggggtaga 601 ggaaacccat tcagacctga tgatgatggt atatttgtaa caagggtaca acctgaagga 661 ccagcatcaa aattactgca gccaggtgat aaaattattc aggctaatgg ctacagtttt 721 ataaatattg aacatggaca agcagtgtcc ttgctaaaaa ctttccagaa tacagttgaa 781 ctcatcattg tacgagaagt tggtaacggt gcaaaacaag agattcgagt gagggttgaa 841 aaagacggtg gttctggagg cgtttcttct gttccgacca atctggaagt tgttgctgcg 901 accccgacta gcctgctgat cagctgggat gcgtactacg actctcatgt ttcttattac 961 cgtatcacgt acggtgaaac cggtggtaac tccccggttc aggaattcac tgtacctggt 1021 tccaagtcta ctgctaccat cagcggcctg aaaccgggtg ttgactatac catcactgta 1081 tacgcacatt acaactacca ttactactct agcccaatct cgattaacta ccgtacctct 1141 agactcgagt catgtaatta gttatgtcac gcttacattc acgccctccc cccacatccg 1201 ctctaaccga aaaggaagga gttagacaac ctgaagtcta ggtccctatt tattttttta 1261 tagttatgtt agtattaaga acgttattta tatttcaaat ttttcttttt tttctgtaca 1321 gacgcgtgta cgcatgtaac attatactga aaaccttgct tgagaaggtt ttgggacgct 1381 cgaaggcttt aatttgcggc cggcgcctga tgcggtattt tctccttacg catctgtgcg 1441 gtatttcaca ccgcatatgg tgcactctca gtacaatctg ctctgatgcc gcatagttaa 1501 gccagccccg acacccgcca acacccgctg acgcgccctg acgggcttgt ctgctcccgg 1561 catccgctta cagacaagct gtgaccgtct ccgggagctg catgtgtcag aggttttcac 1621 cgtcatcacc gaaacgcgcg agacgaaagg gcgggtaata actgatataa ttaaattgaa 1681 gctctaattt gtgagtttag tatacatgca tttacttata atacagtttt ttagttttgc 1741 tggccgcatc ttctcaaata tgcttcccag cctgcttttc tgtaacgttc accctctacc 1801 ttagcatccc ttccctttgc aaatagtcct cttccaacaa taataatgtc agatcctgta 1861 gagaccacat catccacggt tctatactgt tgacccaatg cgtctccctt gtcatctaaa 1921 cccacaccgg gtgtcataat caaccaatcg taaccttcat ctcttccacc catgtctctt 1981 tgagcaataa agccgataac aaaatctttg tcgctcttcg caatgtcaac agtaccctta 2041 gtatattctc cagtagatag ggagcccttg catgacaatt ctgctaacat caaaaggcct 2101 ctaggttcct ttgttacttc ttctgccgcc tgcttcaaac cgctaacaat acctgggccc 2161 accacaccgt gtgcattcgt aatgtctgcc cattctgcta ttctgtatac acccgcagag 2221 tactgcaatt tgactgtatt accaatgtca gcaaattttc tgtcttcgaa gagtaaaaaa 2281 ttgtacttgg cggataatgc ctttagcggc ttaactgtgc cctccatgga aaaatcagtc 2341 aagatatcca catgtgtttt tagtaaacaa attttgggac ctaatgcttc aactaactcc 2401 agtaattcct tggtggtacg aacatccaat gaagcacaca agtttgtttg cttttcgtgc 2461 atgatattaa atagcttggc agcaacagga ctaggatgag tagcagcacg ttccttatat 2521 gtagctttcg acatgattta tcttcgtttc ctgcatgttt ttgttctgtg cagttgggtt 2581 aagaatactg ggcaatttca tgtttcttca acactacata tgcgtatata taccaatcta 2641 agtctgtgct ccttccttcg ttcttccttc tgttcggaga ttaccgaatc aaaaaaattt 2701 caaagaaacc gaaatcaaaa aaaagaataa aaaaaaaatg atgaattgaa ttgaaaagct 2761 ggcctcgtga tacgcctatt tttataggtt aatgtcatga taataatggt ttcttagacg 2821 tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata 2881 cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga 2941 aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc ttattccctt ttttgcggca 3001 ttttgccttc ctgtttttgc tcacccagaa acgctggtga aagtaaaaga tgctgaagat 3061 cagttgggtg cacgagtggg ttacatcgaa ctggatctca acagcggtaa gatccttgag 3121 agttttcgcc ccgaagaacg ttttccaatg atgagcactt ttaaagttct gctatgtggc 3181 gcggtattat cccgtattga cgccgggcaa gagcaactcg gtcgccgcat acactattct 3241 cagaatgact tggttgagta ctcaccagtc acagaaaagc atcttacgga tggcatgaca 3301 gtaagagaat tatgcagtgc tgccataacc atgagtgata acactgcggc caacttactt 3361 ctgacaacga tcggaggacc gaaggagcta accgcttttt tgcacaacat gggggatcat 3421 gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt 3481 gacaccacga tgcctgtagc aatggcaaca acgttgcgca aactattaac tggcgaacta 3541 cttactctag cttcccggca acaattaata gactggatgg aggcggataa agttgcagga 3601 ccacttctgc gctcggccct tccggctggc tggtttattg ctgataaatc tggagccggt 3661 gagcgtgggt ctcgcggtat cattgcagca ctggggccag atggtaagcc ctcccgtatc 3721 gtagttatct acacgacggg gagtcaggca actatggatg aacgaaatag acagatcgct 3781 gagataggtg cctcactgat taagcattgg taactgtcag accaagttta ctcatatata 3841 ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt 3901 gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc 3961 gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg 4021 caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact 4081 ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt ccttctagtg 4141 tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg 4201 ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac 4261 tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca 4321 cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga 4381 gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc 4441 ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct 4501 gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg 4561 agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct 4621 tttgctcaca tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc 4681 tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc 4741 gaggaagcgg aagagcgccc aatacgcaaa ccgcctctcc ccgcgcgttg gccgattcat 4801 taatgcagct ggcacgacag gtttcccgac tggaaagcgg gcagtgagcg caacgcaatt 4861 aatgtgagtt agctcactca ttaggcaccc caggctttac actttatgct tccggctcgt 4921 atgttgtgtg gaattgtgag cggataacaa tttcacacag gaaacagcta tgaccatgat 4981 tacgccaagc taattaaccc tcactaaagg gaacaaaagc tggagct //