LOCUS Exported 4290 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS pTF270 (pFA6a-PSc-6xGly-3xFlag-KanMX) SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 4290) TITLE Formosa lab yeast 3' tagging constructs JOURNAL Unpublished REFERENCE 2 (bases 1 to 4290) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..4290 /organism="synthetic DNA construct" /mol_type="other DNA" promoter complement(199..303) /gene="bla" /label=AmpR promoter primer_bind 371..389 /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" primer_bind complement(427..449) /label=pGEX 3' /note="pGEX vectors, reverse primer" primer_bind 549..568 /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" promoter 649..667 /label=SP6 promoter /note="promoter for bacteriophage SP6 RNA polymerase" primer_bind 649..666 /label=SP6 /note="SP6 promoter, forward primer" CDS 728..751 /codon_start=1 /product="recognition and cleavage site for human rhinovirus 3C and PreScission proteases" /label=HRV 3C site /translation="LEVLFQGP" CDS 776..841 /codon_start=1 /product="three tandem FLAG(R) epitope tags, followed by an enterokinase cleavage site" /label=3xFLAG /translation="DYKDHDGDYKDHDIDYKDDDDK" terminator 870..1057 /gene="S. cerevisiae ADH1" /label=ADH1 terminator /note="transcription terminator for the S. cerevisiae alcohol dehydrogenase 1 (ADH1) gene" primer_bind complement(925..949) /label=pHybLex-R /note="ADH terminator, reverse primer" gene 1132..2488 /label=kanMX /note="yeast selectable marker conferring kanamycin resistance (Wach et al., 1994)" promoter 1132..1475 /label=TEF promoter /note="Ashbya gossypii TEF promoter" CDS 1476..2285 /codon_start=1 /gene="aph(3')-Ia" /product="aminoglycoside phosphotransferase" /label=KanR /note="confers resistance to kanamycin" /translation="MGKEKTHVSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF" primer_bind complement(1543..1562) /label=Kan-R /note="Kanamycin resistance gene, reverse primer" terminator 2291..2488 /label=TEF terminator /note="Ashbya gossypii TEF terminator" primer_bind complement(2592..2611) /label=T7 /note="T7 promoter, forward primer" promoter complement(2593..2611) /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" primer_bind complement(2698..2715) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(2869..3457) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(2949..2968) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(3628..198) /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 4251..4270 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" ORIGIN 1 cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag 61 ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt 121 ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg 181 gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta 241 ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc 301 gcgcacattt ccccgaaaag tgccacctga cgtctaagaa accattatta tcatgacatt 361 aacctataaa aataggcgta tcacgaggcc ctttcgtctc gcgcgtttcg gtgatgacgg 421 tgaaaacctc tgacacatgc agctcccgga gacggtcaca gcttgtctgt aagcggatgc 481 cgggagcaga caagcccgtc agggcgcgtc agcgggtgtt ggcgggtgtc ggggctggct 541 taactatgcg gcatcagagc agattgtact gagagtgcac catatggaca tattgtcgtt 601 agaacgcggc tacaattaat acataacctt atgtatcata cacatacgat ttaggtgaca 661 ctatagaacg cggccgccag ctgaagcttc gtacgctgca ggtcgacgga tccccgggtt 721 aattaacttg gaagtattgt tccaaggtcc aggtggaggc ggtggaggct ctagagacta 781 caaggaccac gacggtgact acaaggacca cgacattgac tacaaggatg acgatgacaa 841 gtgactcgag gcgcgccact tctaaataag cgaatttctt atgatttatg atttttatta 901 ttaaataagt tataaaaaaa ataagtgtat acaaatttta aagtgactct taggttttaa 961 aacgaaaatt cttattcttg agtaactctt tcctgtaggt caggttgctt tctcaggtat 1021 agtatgaggt cgctcttatt gaccacacct ctaccggcag atccgctagg gataacaggg 1081 taatatagat ctgtttagct tgcctcgtcc ccgccgggtc acccggccag cgacatggag 1141 gcccagaata ccctccttga cagtcttgac gtgcgcagct caggggcatg atgtgactgt 1201 cgcccgtaca tttagcccat acatccccat gtataatcat ttgcatccat acattttgat 1261 ggccgcacgg cgcgaagcaa aaattacggc tcctcgctgc agacctgcga gcagggaaac 1321 gctcccctca cagacgcgtt gaattgtccc cacgccgcgc ccctgtagag aaatataaaa 1381 ggttaggatt tgccactgag gttcttcttt catatacttc cttttaaaat cttgctagga 1441 tacagttctc acatcacatc cgaacataaa caaccatggg taaggaaaag actcacgttt 1501 cgaggccgcg attaaattcc aacatggatg ctgatttata tgggtataaa tgggctcgcg 1561 ataatgtcgg gcaatcaggt gcgacaatct atcgattgta tgggaagccc gatgcgccag 1621 agttgtttct gaaacatggc aaaggtagcg ttgccaatga tgttacagat gagatggtca 1681 gactaaactg gctgacggaa tttatgcctc ttccgaccat caagcatttt atccgtactc 1741 ctgatgatgc atggttactc accactgcga tccccggcaa aacagcattc caggtattag 1801 aagaatatcc tgattcaggt gaaaatattg ttgatgcgct ggcagtgttc ctgcgccggt 1861 tgcattcgat tcctgtttgt aattgtcctt ttaacagcga tcgcgtattt cgtctcgctc 1921 aggcgcaatc acgaatgaat aacggtttgg ttgatgcgag tgattttgat gacgagcgta 1981 atggctggcc tgttgaacaa gtctggaaag aaatgcataa gcttttgcca ttctcaccgg 2041 attcagtcgt cactcatggt gatttctcac ttgataacct tatttttgac gaggggaaat 2101 taataggttg tattgatgtt ggacgagtcg gaatcgcaga ccgataccag gatcttgcca 2161 tcctatggaa ctgcctcggt gagttttctc cttcattaca gaaacggctt tttcaaaaat 2221 atggtattga taatcctgat atgaataaat tgcagtttca tttgatgctc gatgagtttt 2281 tctaatcagt actgacaata aaaagattct tgttttcaag aacttgtcat ttgtatagtt 2341 tttttatatt gtagttgttc tattttaatc aaatgttagc gtgatttata ttttttttcg 2401 cctcgacatc atctgcccag atgcgaagtt aagtgcgcag aaagtaatat catgcgtcaa 2461 tcgtatgtga atgctggtcg ctatactgct gtcgattcga tactaacgcc gccatccagt 2521 ttaaacgagc tcgaattcat cgatgatatc agatccacta gtggcctatg cggccgcgga 2581 tctgccggtc tccctatagt gagtcgtatt aatttcgata agccaggtta acctgcatta 2641 atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc 2701 gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa 2761 ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa 2821 aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct 2881 ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac 2941 aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc 3001 gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc 3061 tcaatgctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg 3121 tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga 3181 gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta acaggattag 3241 cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta 3301 cactagaagg acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag 3361 agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg 3421 caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac 3481 ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc 3541 aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag 3601 tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc 3661 agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac 3721 gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc 3781 accggctcca gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg 3841 tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag 3901 tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc 3961 acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac 4021 atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag 4081 aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac 4141 tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg 4201 agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc 4261 gccacatagc agaactttaa aagtgctcat //