LOCUS Exported 4530 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS pTF280 (pFA6a-PSc-6xGly-2xStrep-HphMX) SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 4530) TITLE Formosa lab yeast 3' tagging constructs JOURNAL Unpublished REFERENCE 2 (bases 1 to 4530) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..4530 /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..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" gene 1150..2725 /label=hphMX6 /note="yeast selectable marker conferring hygromycin resistance" promoter 1150..1493 /label=TEF promoter /note="Ashbya gossypii TEF promoter" CDS 1494..2522 /codon_start=1 /gene="aph(4)-Ia" /product="aminoglycoside phosphotransferase from E. coli" /label=HygR /note="confers resistance to hygromycin" /translation="MGKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGR GYVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDL PETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHW QTVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAM FGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVD GNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRA KE" terminator 2528..2725 /label=TEF terminator /note="Ashbya gossypii TEF terminator" primer_bind complement(2832..2851) /label=T7 /note="T7 promoter, forward primer" promoter complement(2833..2851) /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" primer_bind complement(2938..2955) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(3109..3697) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(3189..3208) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(3868..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 4491..4510 /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 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 ctcgctgcgg 1321 acctgcgagc agggaaacgc tcccctcaca gacgcgttga attgtcccca cgccgcgccc 1381 ctgtagagaa atataaaagg ttaggatttg ccactgaggt tcttctttca tatacttcct 1441 tttaaaatct tgctaggata cagttctcac atcacatccg aacataaaca accatgggta 1501 aaaagcctga actcaccgcg acgtctgtcg agaagtttct gatcgaaaag ttcgacagcg 1561 tctccgacct gatgcagctc tcggagggcg aagaatctcg tgctttcagc ttcgatgtag 1621 gagggcgtgg atatgtcctg cgggtaaata gctgcgccga tggtttctac aaagatcgtt 1681 atgtttatcg gcactttgca tcggccgcgc tcccgattcc ggaagtgctt gacattgggg 1741 aattcagcga gagcctgacc tattgcatct cccgccgtgc acagggtgtc acgttgcaag 1801 acctgcctga aaccgaactg cccgctgttc tgcagccggt cgcggaggcc atggatgcga 1861 tcgctgcggc cgatcttagc cagacgagcg ggttcggccc attcggaccg caaggaatcg 1921 gtcaatacac tacatggcgt gatttcatat gcgcgattgc tgatccccat gtgtatcact 1981 ggcaaactgt gatggacgac accgtcagtg cgtccgtcgc gcaggctctc gatgagctga 2041 tgctttgggc cgaggactgc cccgaagtcc ggcacctcgt gcacgcggat ttcggctcca 2101 acaatgtcct gacggacaat ggccgcataa cagcggtcat tgactggagc gaggcgatgt 2161 tcggggattc ccaatacgag gtcgccaaca tcttcttctg gaggccgtgg ttggcttgta 2221 tggagcagca gacgcgctac ttcgagcgga ggcatccgga gcttgcagga tcgccgcggc 2281 tccgggcgta tatgctccgc attggtcttg accaactcta tcagagcttg gttgacggca 2341 atttcgatga tgcagcttgg gcgcagggtc gatgcgacgc aatcgtccga tccggagccg 2401 ggactgtcgg gcgtacacaa atcgcccgca gaagcgcggc cgtctggacc gatggctgtg 2461 tagaagtact cgccgatagt ggaaaccgac gccccagcac tcgtccgagg gcaaaggaat 2521 aatcagtact gacaataaaa agattcttgt tttcaagaac ttgtcatttg tatagttttt 2581 ttatattgta gttgttctat tttaatcaaa tgttagcgtg atttatattt tttttcgcct 2641 cgacatcatc tgcccagatg cgaagttaag tgcgcagaaa gtaatatcat gcgtcaatcg 2701 tatgtgaatg ctggtcgcta tactgctgtc gattcgatac taacgccgcc atccagtgtc 2761 gaaaacgagc tcgaattcat cgatgatatc agatccacta gtggcctatg cggccgcgga 2821 tctgccggtc tccctatagt gagtcgtatt aatttcgata agccaggtta acctgcatta 2881 atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc 2941 gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa 3001 ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa 3061 aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct 3121 ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac 3181 aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc 3241 gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc 3301 tcaatgctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg 3361 tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga 3421 gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta acaggattag 3481 cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta 3541 cactagaagg acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag 3601 agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg 3661 caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac 3721 ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc 3781 aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag 3841 tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc 3901 agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac 3961 gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc 4021 accggctcca gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg 4081 tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag 4141 tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc 4201 acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac 4261 atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag 4321 aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac 4381 tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg 4441 agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc 4501 gccacatagc agaactttaa aagtgctcat //