LOCUS Exported 6418 bp ds-DNA circular SYN 14-AUG-2016DEFINITION synthetic circular DNAKEYWORDS pYC25SOURCE synthetic DNA construct ORGANISM synthetic DNA constructREFERENCE 1 (bases 1 to 6418) AUTHORS Biofeng Lab. TITLE Direct SubmissionFEATURES Location/Qualifiers source 1..6418 /organism="synthetic DNA construct" /mol_type="other DNA" promoter 666..737 /gene="bla" /note="AmpR promoter" CDS 738..1598 /codon_start=1 /gene="bla" /product="beta-lactamase" /note="AmpR" /note="confers resistance to ampicillin, carbenicillin, and related antibiotics" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS LIKHW" rep_origin 1753..2372 /direction=RIGHT /note="pBR322" /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" promoter 2681..2711 /note="lac promoter" /note="promoter for the E. coli lac operon" protein_bind 2719..2735 /bound_moiety="lac repressor encoded by lacI" /note="lac operator" /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 2743..2759 /note="M13 rev" /note="common sequencing primer, one of multiple similar variants" promoter 2780..2798 /note="T3 promoter" /note="promoter for bacteriophage T3 RNA polymerase" promoter 2816..3214 /gene="S. cerevisiae TEF1" /note="TEF1 promoter" /note="promoter for EF-1-alpha" misc_feature 3218..3235 /note="MCS" primer_bind 3218..3234 /note="SK primer" /note="common sequencing primer, one of multiple similar variants" CDS 3238..3954 /codon_start=1 /note="YFP" /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK LICTTGKLPVPWPTLVTTLGYGLMCFARYPDHMKRHDFFKSAMPEGYVQERTIFFKDDG NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIKA NFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLLE FVTAAGITHGMDELYK" primer_bind complement(3993..4009) /note="KS primer" /note="common sequencing primer, one of multiple similar variants" promoter complement(4424..4442) /note="T7 promoter" /note="promoter for bacteriophage T7 RNA polymerase" primer_bind complement(4452..4468) /note="M13 fwd" /note="common sequencing primer, one of multiple similar variants" rep_origin 4613..5068 /direction=RIGHT /note="f1 ori" /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" CDS complement(5199..6002) /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)" /translation="MSKATYKERAATHPSPVAAKLFNIMHEKQTNLCASLDVRTTKELL ELVEALGPKICLLKTHVDILTDFSMEGTVKPLKALSAKYNFLLFEDRKFADIGNTVKLQ YSAGVYRIAEWADITNAHGVVGPGIVSGLKQAAEEVTKEPRGLLMLAELSCKGSLSTGE YTKGTVDIAKSDKDFVIGFIAQRDMGGRDEGYDWLIMTPGVGLDDKGDALGQQYRTVDD VVSTGSDIIIVGRGLFAKGRDAKVEGERYRKAGWEAYLRRCGQQN" promoter complement(6003..6218) /gene="S. cerevisiae URA3" /note="URA3 promoter"ORIGIN 1 gacgaaaggg cctcgtgata cgcctatttt tataggttaa tgtcatgata ataatggttt 61 cttagcaggt tagactaaca tgcaaaaagt ttatattcat ttttgtagtt attcttgttt 121 tcattaggat gattacacac tgtggatgtg gattaatcga tcattgttaa tttagtgtag 181 ctactaccat ttcaaacaaa acgtaaattc ttgcagtcgt actggatctg tgaatctatt 241 agtatatatg aattaaagta gcttgtacat attattctgt tgaatcatat cgcagagcat 301 tggttgaaat ccaaaatata aaaatgtaat atcacaaaaa ataatactaa ttctaacatt 361 aatgggtcag atttttagtg aatacttaaa tttataatat cgtctattta agctagcaaa 421 tggaacaaca tttaaagtaa gaacatcata tctacatgaa aatgtatatt tcaatctgac 481 taataacgca gagcacatct ttcagtgatg tctgtcacat gatcaaaaag aattgtattt 541 aaataattca taataaaagc ttaaaaaatt acaataatga aaataaagta aataatgaca 601 tgggtaagag ttccgaaata gaatcttagt gtacaaaaca aaaattgcat cattagagat 661 ccccacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat 721 attgaaaaag gaagagtatg agtattcaac atttccgtgt cgcccttatt cccttttttg 781 cggcattttg ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta aaagatgctg 841 aagatcagtt gggtgcacga gtgggttaca tcgaactgga tctcaacagc ggtaagatcc 901 ttgagagttt tcgccccgaa gaacgttttc caatgatgag cacttttaaa gttctgctat 961 gtggcgcggt attatcccgt attgacgccg ggcaagagca actcggtcgc cgcatacact 1021 attctcagaa tgacttggtt gagtactcac cagtcacaga aaagcatctt acggatggca 1081 tgacagtaag agaattatgc agtgctgcca taaccatgag tgataacact gcggccaact 1141 tacttctgac aacgatcgga ggaccgaagg agctaaccgc ttttttgcac aacatggggg 1201 atcatgtaac tcgccttgat cgttgggaac cggagctgaa tgaagccata ccaaacgacg 1261 agcgtgacac cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg 1321 aactacttac tctagcttcc cggcaacaat taatagactg gatggaggcg gataaagttg 1381 caggaccact tctgcgctcg gcccttccgg ctggctggtt tattgctgat aaatctggag 1441 ccggtgagcg tgggtctcgc ggtatcattg cagcactggg gccagatggt aagccctccc 1501 gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga 1561 tcgctgagat aggtgcctca ctgattaagc attggtaact gtcagaccaa gtttactcat 1621 atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc 1681 tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag 1741 accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct 1801 gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac 1861 caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat actgtccttc 1921 tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg 1981 ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt 2041 tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt 2101 gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc 2161 tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca 2221 gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata 2281 gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg 2341 ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct 2401 ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta 2461 ccgcctttga gtgagctgat accgctcgcc gcagccgaac gaccgagcgc agcgagtcag 2521 tgagcgagga agcggaagag cgcccaatac gcaaaccgcc tctccccgcg cgttggccga 2581 ttcattaatg cagctggcac gacaggtttc ccgactggaa agcgggcagt gagcgcaacg 2641 caattaatgt gagttacctc actcattagg caccccaggc tttacacttt atgcttccgg 2701 ctcctatgtt gtgtggaatt gtgagcggat aacaatttca cacaggaaac agctatgacc 2761 atgattacgc caagcgcgca attaaccctc actaaaggga acaaaagctg gagctcatag 2821 cttcaaaatg tttctactcc ttttttactc ttccagattt tctcggactc cgcgcatcgc 2881 cgtaccactt caaaacaccc aagcacagca tactaaattt cccctctttc ttcctctagg 2941 gtgtcgttaa ttacccgtac taaaggtttg gaaaagaaaa aagagaccgc ctcgtttctt 3001 tttcttcgtc gaaaaaggca ataaaaattt ttatcacgtt tctttttctt gaaaattttt 3061 tttttgattt ttttctcttt cgatgacctc ccattgatat ttaagttaat aaacggtctt 3121 caatttctca agtttcagtt tcatttttct tgttctatta caactttttt tacttcttgc 3181 tcattagaaa gaaagcatag caatctaatc taagttttct agaactagtg gatcctaatg 3241 agtaaaggag aagaactttt cactggagtt gtcccaattc ttgttgaatt agatggtgat 3301 gttaatgggc acaaattttc tgtcagtgga gagggtgaag gtgatgcaac atacggaaaa 3361 cttaccctta aattgatttg cactactgga aaactacctg ttccatggcc aacacttgtc 3421 actactttgg gttatggtct aatgtgcttt gctagatacc cagatcatat gaaacggcat 3481 gactttttca agagtgccat gcccgaaggt tatgtacagg aaagaactat atttttcaaa 3541 gatgacggga actacaagac acgtgctgaa gtcaagtttg aaggtgatac ccttgttaat 3601 agaatcgagt taaaaggtat tgattttaaa gaagatggaa acattcttgg acacaaattg 3661 gaatacaact ataactcaca caatgtatac atcactgcag acaaacaaaa gaatggaatc 3721 aaagctaact tcaaaattag acacaacatt gaagatggag gtgttcaact agcagaccat 3781 tatcaacaaa atactccaat tggcgatggc cctgtccttt taccagacaa ccattacctg 3841 tcctatcaat ctgccctttc gaaagatccc aacgaaaaga gagaccacat ggtccttctt 3901 gagtttgtaa cagctgctgg gattacacat ggcatggatg aactatacaa atagagatcc 3961 cccgggctgc aggaattcga tatcaagctt atcgataccg tcgacctcga gaacacaacc 4021 cacagctacc accatcaaca atatttatat atataacgta cacatagaaa tcacacaaac 4081 agagtattta ttcttaacta catgaactac catcagaccg tctgggccac tatataatgt 4141 gccattcata aacgtgatca ctttacgtag caggcaaccc caggtgaaaa tttttcagcg 4201 agctgccaga ttgtcaggtg aaaaactgaa aaaaacttct gggcgatgag cttgtggcgg 4261 gaaattaagt atataaagca gttagtttct tctgcttctt gtggttctgg gtattcttgt 4321 aatatttgga agaatagaaa atcaagacta ctgcctttct tttcatatta tagaggaaga 4381 aatacgcacg aacacgatat agaggtaaag gtacccaatt cgccctatag tgagtcgtat 4441 tacgcgcgct cactggccgt cgttttacaa cgtcgtgact gggaaaaccc tggcgttacc 4501 caacttaatc gccttgcagc acatccccct ttcgccagct ggcgtaatag cgaagaggcc 4561 cgcaccgatc gcccttccca acagttgcgc agcctgaatg gcgaatggcg cgacgcgccc 4621 tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt 4681 gccagcgccc tagcgcccgc tcctttcgct ttcttccctt cctttctcgc cacgttcgcc 4741 ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt tagtgcttta 4801 cggcacctcg accccaaaaa acttgattag ggtgatggtt cacgtagtgg gccatcgccc 4861 tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag tggactcttg 4921 ttccaaactg gaacaacact caaccctatc tcggtctatt cttttgattt ataagggatt 4981 ttgccgattt cggcctattg gttaaaaaat gagctgattt aacaaaaatt taacgcgaat 5041 tttaacaaaa tattaacgtt tacaatttcc tgatgcggta ttttctcctt acgcatctgt 5101 gcggtatttc acaccgcata gggtaataac tgatataatt aaattgaagc tctaatttgt 5161 gagtttagta tacatgcatt tacttataat acagtttttt agttttgctg gccgcatctt 5221 ctcaaatatg cttcccagcc tgcttttctg taacgttcac cctctacctt agcatccctt 5281 ccctttgcaa atagtcctct tccaacaata ataatgtcag atcctgtaga gaccacatca 5341 tccacggttc tatactgttg acccaatgcg tctcccttgt catctaaacc cacaccgggt 5401 gtcataatca accaatcgta accttcatct cttccaccca tgtctctttg agcaataaag 5461 ccgataacaa aatctttgtc gctcttcgca atgtcaacag tacccttagt atattctcca 5521 gtagataggg agcccttgca tgacaattct gctaacatca aaaggcctct aggttccttt 5581 gttacttctt ctgccgcctg cttcaaaccg ctaacaatac ctgggcccac cacaccgtgt 5641 gcattcgtaa tgtctgccca ttctgctatt ctgtatacac ccgcagagta ctgcaatttg 5701 actgtattac caatgtcagc aaattttctg tcttcgaaga gtaaaaaatt gtacttggcg 5761 gataatgcct ttagcggctt aactgtgccc tccatggaaa aatcagtcaa gatatccaca 5821 tgtgttttta gtaaacaaat tttgggacct aatgcttcaa ctaactccag taattccttg 5881 gtggtacgaa catccaatga agcacacaag tttgtttgct tttcgtgcat gatattaaat 5941 agcttggcag caacaggact aggatgagta gcagcacgtt ccttatatgt agctttcgac 6001 atgatttatc ttcgtttcct gcaggttttt gttctgtgca gttgggttaa gaatactggg 6061 caatttcatg tttcttcaac actacatatg cgtatatata ccaatctaag tctgtgctcc 6121 ttccttcgtt cttccttctg ttcggagatt accgaatcaa aaaaatttca aagaaaccga 6181 aatcaaaaaa aagaataaaa aaaaaatgat gaattgaatt gaaaagctgt ggtatggtgc 6241 actctcagta caatctgctc tgatgccgca tagttaagcc agccccgaca cccgccaaca 6301 cccgctgacg cgccctgacg ggcttgtctg ctcccggcat ccgcttacag acaagctgtg 6361 accgtctccg ggaggtgcat gtgtcagagg ttttcaccgt catcaccgaa acgcgcga