LOCUS Exported 4512 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS pTF272 (pFA6a-TEV-6xGly-3xFlag-HphMX) SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 4512) TITLE Formosa lab yeast 3' tagging constructs JOURNAL Unpublished REFERENCE 2 (bases 1 to 4512) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..4512 /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..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..2707 /label=hphMX6 /note="yeast selectable marker conferring hygromycin resistance" promoter 1132..1475 /label=TEF promoter /note="Ashbya gossypii TEF promoter" CDS 1476..2504 /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 2510..2707 /label=TEF terminator /note="Ashbya gossypii TEF terminator" primer_bind complement(2814..2833) /label=T7 /note="T7 promoter, forward primer" promoter complement(2815..2833) /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" primer_bind complement(2920..2937) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(3091..3679) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(3171..3190) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(3850..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 4473..4492 /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 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 taaaaagcct gaactcaccg 1501 cgacgtctgt cgagaagttt ctgatcgaaa agttcgacag cgtctccgac ctgatgcagc 1561 tctcggaggg cgaagaatct cgtgctttca gcttcgatgt aggagggcgt ggatatgtcc 1621 tgcgggtaaa tagctgcgcc gatggtttct acaaagatcg ttatgtttat cggcactttg 1681 catcggccgc gctcccgatt ccggaagtgc ttgacattgg ggaattcagc gagagcctga 1741 cctattgcat ctcccgccgt gcacagggtg tcacgttgca agacctgcct gaaaccgaac 1801 tgcccgctgt tctgcagccg gtcgcggagg ccatggatgc gatcgctgcg gccgatctta 1861 gccagacgag cgggttcggc ccattcggac cgcaaggaat cggtcaatac actacatggc 1921 gtgatttcat atgcgcgatt gctgatcccc atgtgtatca ctggcaaact gtgatggacg 1981 acaccgtcag tgcgtccgtc gcgcaggctc tcgatgagct gatgctttgg gccgaggact 2041 gccccgaagt ccggcacctc gtgcacgcgg atttcggctc caacaatgtc ctgacggaca 2101 atggccgcat aacagcggtc attgactgga gcgaggcgat gttcggggat tcccaatacg 2161 aggtcgccaa catcttcttc tggaggccgt ggttggcttg tatggagcag cagacgcgct 2221 acttcgagcg gaggcatccg gagcttgcag gatcgccgcg gctccgggcg tatatgctcc 2281 gcattggtct tgaccaactc tatcagagct tggttgacgg caatttcgat gatgcagctt 2341 gggcgcaggg tcgatgcgac gcaatcgtcc gatccggagc cgggactgtc gggcgtacac 2401 aaatcgcccg cagaagcgcg gccgtctgga ccgatggctg tgtagaagta ctcgccgata 2461 gtggaaaccg acgccccagc actcgtccga gggcaaagga ataatcagta ctgacaataa 2521 aaagattctt gttttcaaga acttgtcatt tgtatagttt ttttatattg tagttgttct 2581 attttaatca aatgttagcg tgatttatat tttttttcgc ctcgacatca tctgcccaga 2641 tgcgaagtta agtgcgcaga aagtaatatc atgcgtcaat cgtatgtgaa tgctggtcgc 2701 tatactgctg tcgattcgat actaacgccg ccatccagtg tcgaaaacga gctcgaattc 2761 atcgatgata tcagatccac tagtggccta tgcggccgcg gatctgccgg tctccctata 2821 gtgagtcgta ttaatttcga taagccaggt taacctgcat taatgaatcg gccaacgcgc 2881 ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg 2941 ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc 3001 cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag 3061 gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca 3121 tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca 3181 ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg 3241 atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcaatgct cacgctgtag 3301 gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt 3361 tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca 3421 cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg 3481 cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa ggacagtatt 3541 tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc 3601 cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg 3661 cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg 3721 gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta 3781 gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg 3841 gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg 3901 ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc 3961 atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc 4021 agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc 4081 ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag 4141 tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat 4201 ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg 4261 caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt 4321 gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag 4381 atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg 4441 accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt 4501 aaaagtgctc at //