LOCUS Exported 3421 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS pFA6a-TRP1 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 3421) AUTHORS Longtine MS, McKenzie A 3rd, Demarini DJ, Shah NG, Wach A, Brachat A, Philippsen P, Pringle JR TITLE Additional modules for versatile and economical PCR-based gene deletion and modification in Saccharomyces cerevisiae. JOURNAL Yeast. 1998 Jul;14(10):953-61. PUBMED 9717241 REFERENCE 2 (bases 1 to 3421) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..3421 /organism="synthetic DNA construct" /mol_type="other DNA" promoter 76..223 /gene="S. cerevisiae TRP1" /label=TRP1 promoter CDS 224..898 /codon_start=1 /gene="S. cerevisiae TRP1" /product="phosphoribosylanthranilate isomerase, required for tryptophan biosynthesis" /label=TRP1 /note="yeast auxotrophic marker" /translation="MSVINFTGSSGPLVKVCGLQSTEAAECALDSDADLLGIICVPNRK RTIDPVIARKISSLVKAYKNSSGTPKYLVGVFRNQPKEDVLALVNDYGIDIVQLHGDES WQEYQEFLGLPVIKRLVFPKDCNILLSAASQKPHSFIPLFDSEAGGTGELLDWNSISDW VGRQESPESLHFMLAGGLTPENVGDALRLNGVIGVDVSGGVETNGVKDSNKIANFVKNA KK" RBS 978..986 /label=Shine-Dalgarno sequence /note="full consensus sequence for ribosome-binding sites upstream of start codons in E. coli; complementary to a region in the 3' end of the 16S rRNA (Chen et al., 1994)" primer_bind complement(1058..1077) /label=T7 /note="T7 promoter, forward primer" promoter complement(1059..1077) /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" primer_bind complement(1164..1181) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(1335..1923) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(1415..1434) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(2094..2954) /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 2717..2736 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(2955..3059) /gene="bla" /label=AmpR promoter primer_bind 3127..3145 /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" primer_bind complement(3183..3205) /label=pGEX 3' /note="pGEX vectors, reverse primer" primer_bind 3305..3324 /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" promoter 3405..2 /label=SP6 promoter /note="promoter for bacteriophage SP6 RNA polymerase" primer_bind 3405..1 /label=SP6 /note="SP6 promoter, forward primer" ORIGIN 1 gaacgcggcc gccagctgaa gcttcgtacg ctgcaggtcg acggatcccc gggttaatta 61 aggcgcgcca gatctgtaca atcttgatcc ggagcttttc tttttttgcc gattaagaat 121 taattcggtc gaaaaaagaa aaggagaggg ccaagaggga gggcattggt gactattgag 181 cacgtgagta tacgtgatta agcacacaaa ggcagcttgg agtatgtctg ttattaattt 241 cacaggtagt tctggtccat tggtgaaagt ttgcggcttg cagagcacag aggccgcaga 301 atgtgctcta gattccgatg ctgacttgct gggtattata tgtgtgccca atagaaagag 361 aacaattgac ccggttattg caaggaaaat ttcaagtctt gtaaaagcat ataaaaatag 421 ttcaggcact ccgaaatact tggttggcgt gtttcgtaat caacctaagg aggatgtttt 481 ggctctggtc aatgattacg gcattgatat cgtccaactg catggagatg agtcgtggca 541 agaataccaa gagttcctcg gtttgccagt tattaaaaga ctcgtatttc caaaagactg 601 caacatacta ctcagtgcag cttcacagaa acctcattcg tttattccct tgtttgattc 661 agaagcaggt gggacaggtg aacttttgga ttggaactcg atttctgact gggttggaag 721 gcaagagagc cccgaaagct tacattttat gttagctggt ggactgacgc cagaaaatgt 781 tggtgatgcg cttagattaa atggcgttat tggtgttgat gtaagcggag gtgtggagac 841 aaatggtgta aaagactcta acaaaatagc aaatttcgtc aaaaatgcta agaaataggt 901 tattactgag tagtatttat ttaagtattg tttgtgcact tgccgatcta tgcggtgtga 961 aataccgcac agatgcgtaa ggaggtttaa acgagctcga attcatcgat gatatcagat 1021 ccactagtgg cctatgcggc cgcggatctg ccggtctccc tatagtgagt cgtattaatt 1081 tcgataagcc aggttaacct gcattaatga atcggccaac gcgcggggag aggcggtttg 1141 cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg 1201 cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat 1261 aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc 1321 gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc 1381 tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga 1441 agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt 1501 ctcccttcgg gaagcgtggc gctttctcaa tgctcacgct gtaggtatct cagttcggtg 1561 taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc 1621 gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg 1681 gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc 1741 ttgaagtggt ggcctaacta cggctacact agaaggacag tatttggtat ctgcgctctg 1801 ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc 1861 gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct 1921 caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt 1981 taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa 2041 aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa 2101 tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc 2161 tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct 2221 gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca 2281 gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt 2341 aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt 2401 gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc 2461 ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc 2521 tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt 2581 atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact 2641 ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc 2701 ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt 2761 ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg 2821 atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct 2881 gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa 2941 tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt 3001 ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc 3061 acatttcccc gaaaagtgcc acctgacgtc taagaaacca ttattatcat gacattaacc 3121 tataaaaata ggcgtatcac gaggcccttt cgtctcgcgc gtttcggtga tgacggtgaa 3181 aacctctgac acatgcagct cccggagacg gtcacagctt gtctgtaagc ggatgccggg 3241 agcagacaag cccgtcaggg cgcgtcagcg ggtgttggcg ggtgtcgggg ctggcttaac 3301 tatgcggcat cagagcagat tgtactgaga gtgcaccata tggacatatt gtcgttagaa 3361 cgcggctaca attaatacat aaccttatgt atcatacaca tacgatttag gtgacactat 3421 a //