LOCUS Exported 4736 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS pDS297 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 4736) AUTHORS Strickland D, Lin Y, Wagner E, Hope CM, Zayner J, Antoniou C, Sosnick TR, Weiss EL, Glotzer M TITLE TULIPs: tunable, light-controlled interacting protein tags for cell biology. JOURNAL Nat Methods. 2012 Mar 4. doi: 10.1038/nmeth.1904. PUBMED 22388287 REFERENCE 2 (bases 1 to 4736) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..4736 /organism="synthetic DNA construct" /mol_type="other DNA" promoter 23..464 /gene="S. cerevisiae GAL1" /label=GAL1 promoter /note="inducible promoter, regulated by Gal4" protein_bind 23..140 /label=UAS /bound_moiety="Gal4" /note="upstream activating sequence mediating Gal4-dependent induction" primer_bind 435..458 /label=GAL1 /note="S. cerevisiae GAL1 promoter, forward primer" primer_bind complement(536..552) /label=KS primer /note="common sequencing primer, one of multiple similar variants" primer_bind complement(537..553) /label=pBluescriptKS /note="For pBluescript vector" terminator 859..1106 /gene="S. cerevisiae CYC1" /label=CYC1 terminator /note="transcription terminator for CYC1" primer_bind complement(875..893) /label=CYC1 /note="CYC1 transcription termination signal, reverse primer" primer_bind complement(1181..1200) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind 1300..1322 /label=pGEX 3' /note="pGEX vectors, reverse primer" CDS complement(1440..2243) /codon_start=1 /gene="S. cerevisiae URA3" /product="orotidine-5'-phosphate decarboxylase, required for uracil biosynthesis" /label=URA3 /note="yeast auxotrophic marker, counterselectable with 5-fluoroorotic acid (5-FOA)" /translation="MSKATYKERAATHPSPVAAKLFNIMHEKQTNLCASLDVRTTKELL ELVEALGPKICLLKTHVDILTDFSMEGTVKPLKALSAKYNFLLFEDRKFADIGNTVKLQ YSAGVYRIAEWADITNAHGVVGPGIVSGLKQAAEEVTKEPRGLLMLAELSCKGSLSTGE YTKGTVDIAKSDKDFVIGFIAQRDMGGRDEGYDWLIMTPGVGLDDKGDALGQQYRTVDD VVSTGSDIIIVGRGLFAKGRDAKVEGERYRKAGWEAYLRRCGQQN" promoter complement(2244..2459) /gene="S. cerevisiae URA3" /label=URA3 promoter primer_bind complement(2471..2489) /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" promoter 2557..2661 /gene="bla" /label=AmpR promoter CDS 2662..3522 /codon_start=1 /gene="bla" /product="beta-lactamase" /label=AmpR /note="confers resistance to ampicillin, carbenicillin, and related antibiotics" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS LIKHW" primer_bind complement(2880..2899) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 3693..4281 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 4182..4201 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 4435..4452 /label=L4440 /note="L4440 vector, forward primer" protein_bind 4569..4590 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." promoter 4605..4635 /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 4643..4659 /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)." primer_bind 4648..4670 /label=M13/pUC Reverse /note="In lacZ gene" primer_bind 4667..4683 /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind 4667..4683 /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" promoter 4701..4719 /label=T3 promoter /note="promoter for bacteriophage T3 RNA polymerase" 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 aaagactcta gactcgagtc atgtaattag ttatgtcacg cttacattca cgccctcccc 901 ccacatccgc tctaaccgaa aaggaaggag ttagacaacc tgaagtctag gtccctattt 961 atttttttat agttatgtta gtattaagaa cgttatttat atttcaaatt tttctttttt 1021 ttctgtacag acgcgtgtac gcatgtaaca ttatactgaa aaccttgctt gagaaggttt 1081 tgggacgctc gaaggcttta atttgcggcc ggcgcctgat gcggtatttt ctccttacgc 1141 atctgtgcgg tatttcacac cgcatatggt gcactctcag tacaatctgc tctgatgccg 1201 catagttaag ccagccccga cacccgccaa cacccgctga cgcgccctga cgggcttgtc 1261 tgctcccggc atccgcttac agacaagctg tgaccgtctc cgggagctgc atgtgtcaga 1321 ggttttcacc gtcatcaccg aaacgcgcga gacgaaaggg cgggtaataa ctgatataat 1381 taaattgaag ctctaatttg tgagtttagt atacatgcat ttacttataa tacagttttt 1441 tagttttgct ggccgcatct tctcaaatat gcttcccagc ctgcttttct gtaacgttca 1501 ccctctacct tagcatccct tccctttgca aatagtcctc ttccaacaat aataatgtca 1561 gatcctgtag agaccacatc atccacggtt ctatactgtt gacccaatgc gtctcccttg 1621 tcatctaaac ccacaccggg tgtcataatc aaccaatcgt aaccttcatc tcttccaccc 1681 atgtctcttt gagcaataaa gccgataaca aaatctttgt cgctcttcgc aatgtcaaca 1741 gtacccttag tatattctcc agtagatagg gagcccttgc atgacaattc tgctaacatc 1801 aaaaggcctc taggttcctt tgttacttct tctgccgcct gcttcaaacc gctaacaata 1861 cctgggccca ccacaccgtg tgcattcgta atgtctgccc attctgctat tctgtataca 1921 cccgcagagt actgcaattt gactgtatta ccaatgtcag caaattttct gtcttcgaag 1981 agtaaaaaat tgtacttggc ggataatgcc tttagcggct taactgtgcc ctccatggaa 2041 aaatcagtca agatatccac atgtgttttt agtaaacaaa ttttgggacc taatgcttca 2101 actaactcca gtaattcctt ggtggtacga acatccaatg aagcacacaa gtttgtttgc 2161 ttttcgtgca tgatattaaa tagcttggca gcaacaggac taggatgagt agcagcacgt 2221 tccttatatg tagctttcga catgatttat cttcgtttcc tgcatgtttt tgttctgtgc 2281 agttgggtta agaatactgg gcaatttcat gtttcttcaa cactacatat gcgtatatat 2341 accaatctaa gtctgtgctc cttccttcgt tcttccttct gttcggagat taccgaatca 2401 aaaaaatttc aaagaaaccg aaatcaaaaa aaagaataaa aaaaaaatga tgaattgaat 2461 tgaaaagctg gcctcgtgat acgcctattt ttataggtta atgtcatgat aataatggtt 2521 tcttagacgt caggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt 2581 ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa 2641 taatattgaa aaaggaagag tatgagtatt caacatttcc gtgtcgccct tattcccttt 2701 tttgcggcat tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat 2761 gctgaagatc agttgggtgc acgagtgggt tacatcgaac tggatctcaa cagcggtaag 2821 atccttgaga gttttcgccc cgaagaacgt tttccaatga tgagcacttt taaagttctg 2881 ctatgtggcg cggtattatc ccgtattgac gccgggcaag agcaactcgg tcgccgcata 2941 cactattctc agaatgactt ggttgagtac tcaccagtca cagaaaagca tcttacggat 3001 ggcatgacag taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc 3061 aacttacttc tgacaacgat cggaggaccg aaggagctaa ccgctttttt gcacaacatg 3121 ggggatcatg taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac 3181 gacgagcgtg acaccacgat gcctgtagca atggcaacaa cgttgcgcaa actattaact 3241 ggcgaactac ttactctagc ttcccggcaa caattaatag actggatgga ggcggataaa 3301 gttgcaggac cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct 3361 ggagccggtg agcgtgggtc tcgcggtatc attgcagcac tggggccaga tggtaagccc 3421 tcccgtatcg tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga 3481 cagatcgctg agataggtgc ctcactgatt aagcattggt aactgtcaga ccaagtttac 3541 tcatatatac tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag 3601 atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg 3661 tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc 3721 tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag 3781 ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtc 3841 cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac 3901 ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc 3961 gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt 4021 tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt 4081 gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc 4141 ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt 4201 tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca 4261 ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt 4321 tgctggcctt ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt 4381 attaccgcct ttgagtgagc tgataccgct cgccgcagcc gaacgaccga gcgcagcgag 4441 tcagtgagcg aggaagcgga agagcgccca atacgcaaac cgcctctccc cgcgcgttgg 4501 ccgattcatt aatgcagctg gcacgacagg tttcccgact ggaaagcggg cagtgagcgc 4561 aacgcaatta atgtgagtta gctcactcat taggcacccc aggctttaca ctttatgctt 4621 ccggctcgta tgttgtgtgg aattgtgagc ggataacaat ttcacacagg aaacagctat 4681 gaccatgatt acgccaagct aattaaccct cactaaaggg aacaaaagct ggagct //