LOCUS Exported 4195 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS pTF275 (pFA6a-TEV-6xGly-2xStrep-His3MX) SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 4195) TITLE Formosa lab yeast 3' tagging constructs JOURNAL Unpublished REFERENCE 2 (bases 1 to 4195) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..4195 /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..748 /codon_start=1 /product="tobacco etch virus (TEV) protease recognition and cleavage site" /label=TEV site /translation="ENLYFQG" CDS 776..799 /codon_start=1 /product="peptide that binds Strep-Tactin(R), an engineered form of streptavidin" /label=Strep-Tag II /translation="WSHPQFEK" CDS 836..859 /codon_start=1 /product="peptide that binds Strep-Tactin(R), an engineered form of streptavidin" /label=Strep-Tag II /translation="WSHPQFEK" terminator 888..1075 /gene="S. cerevisiae ADH1" /label=ADH1 terminator /note="transcription terminator for the S. cerevisiae alcohol dehydrogenase 1 (ADH1) gene" primer_bind complement(943..967) /label=pHybLex-R /note="ADH terminator, reverse primer" promoter 1150..1493 /label=TEF promoter /note="Ashbya gossypii TEF promoter" terminator 2196..2393 /label=TEF terminator /note="Ashbya gossypii TEF terminator" primer_bind complement(2497..2516) /label=T7 /note="T7 promoter, forward primer" promoter complement(2498..2516) /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" primer_bind complement(2603..2620) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(2774..3362) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(2854..2873) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(3533..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 4156..4175 /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 aattaacgag aatttatact tccaaggttc tggtggaggc ggtggaggct ctagatggtc 781 tcacccacag ttcgagaagg gtggaggctc tggcggtgga tccggaggcg gttcatggtc 841 tcacccacag ttcgagaagt gactcgaggc gcgccacttc taaataagcg aatttcttat 901 gatttatgat ttttattatt aaataagtta taaaaaaaat aagtgtatac aaattttaaa 961 gtgactctta ggttttaaaa cgaaaattct tattcttgag taactctttc ctgtaggtca 1021 ggttgctttc tcaggtatag tatgaggtcg ctcttattga ccacacctct accggcagat 1081 ccgctaggga taacagggta atatagatct gtttagcttg cctcgtcccc gccgggtcac 1141 ccggccagcg acatggaggc ccagaatacc ctccttgaca gtcttgacgt gcgcagctca 1201 ggggcatgat gtgactgtcg cccgtacatt tagcccatac atccccatgt ataatcattt 1261 gcatccatac attttgatgg ccgcacggcg cgaagcaaaa attacggctc ctcgctgcag 1321 acctgcgagc agggaaacgc tcccctcaca gacgcgttga attgtcccca cgccgcgccc 1381 ctgtagagaa atataaaagg ttaggatttg ccactgaggt tcttctttca tatacttcct 1441 tttaaaatct tgctaggata cagttctcac atcacatccg aacataaaca accatggcag 1501 aaccagccca aaaaaagcaa aaacaaactg ttcaggagcg caaggcgttt atctcccgta 1561 tcactaatga aactaaaatt caaatcgcta tttcgctgaa tggtggttat attcaaataa 1621 aagattcgat tcttcctgca aagaaggatg acgatgtagc ttcccaagct actcagtcac 1681 aggtcatcga tattcacaca ggtgttggct ttttggatca tatgatccat gcgttggcaa 1741 aacactctgg ttggtctctt attgttgaat gtatcggtga cctgcacatt gacgatcacc 1801 atactaccga agattgcggt atcgcattag ggcaagcgtt caaagaagca atgggtgctg 1861 tccgtggtgt aaaaagattc ggtactgggt tcgcaccatt ggatgaggcg ctatcacgtg 1921 ccgtagtcga tttatctaat agaccatttg ctgtaatcga ccttggattg aagagagaga 1981 tgattggtga tttatccact gaaatgattc cacacttttt ggaaagtttc gcggaggcgg 2041 ccagaattac tttgcatgtt gattgtctga gaggtttcaa cgatcaccac agaagtgaga 2101 gtgcgttcaa ggctttggct gttgccataa gagaagctat ttctagcaat ggcaccaatg 2161 acgttccctc aaccaaaggt gttttgatgt gaagtactga caataaaaag attcttgttt 2221 tcaagaactt gtcatttgta tagttttttt atattgtagt tgttctattt taatcaaatg 2281 ttagcgtgat ttatattttt tttcgcctcg acatcatctg cccagatgcg aagttaagtg 2341 cgcagaaagt aatatcatgc gtcaatcgta tgtgaatgct ggtcgctata ctgctgtcga 2401 ttcgatacta acgccgccat ccagtttaaa cgagctcgaa ttcatcgatg atatcagatc 2461 cactagtggc ctatgcggcc gcggatctgc cggtctccct atagtgagtc gtattaattt 2521 cgataagcca ggttaacctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc 2581 gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc 2641 ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata 2701 acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg 2761 cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct 2821 caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa 2881 gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc 2941 tcccttcggg aagcgtggcg ctttctcaat gctcacgctg taggtatctc agttcggtgt 3001 aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg 3061 ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg 3121 cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct 3181 tgaagtggtg gcctaactac ggctacacta gaaggacagt atttggtatc tgcgctctgc 3241 tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg 3301 ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc 3361 aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt 3421 aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa 3481 aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat 3541 gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct 3601 gactccccgt cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg 3661 caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag 3721 ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta 3781 attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg 3841 ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg 3901 gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct 3961 ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta 4021 tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg 4081 gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc 4141 cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg ctcat //