LOCUS Exported 4279 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS pRSII404 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 4279) AUTHORS Chee MK, Haase SB TITLE New and Redesigned pRS Plasmid Shuttle Vectors for Genetic Manipulation of Saccharomycescerevisiae. JOURNAL G3 (Bethesda). 2012 May;2(5):515-26. Epub 2012 May 1. PUBMED 22670222 REFERENCE 2 (bases 1 to 4279) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..4279 /organism="synthetic DNA construct" /mol_type="other DNA" primer_bind complement(29..51) /label=pGEX 3' /note="pGEX vectors, reverse primer" primer_bind 151..170 /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" promoter 191..472 /gene="S. cerevisiae TRP1" /label=TRP1 promoter CDS 473..1147 /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" rep_origin complement(1250..1705) /direction=LEFT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(1387..1408) /label=F1ori-F /note="F1 origin, forward primer" primer_bind 1599..1618 /label=F1ori-R /note="F1 origin, reverse primer" CDS complement(1682..2062) /codon_start=1 /gene="lacZ fragment" /product="LacZ-alpha fragment of beta-galactosidase" /label=lacZ-alpha /translation="MTMITPSAQLTLTKGNKSWSSTAVAAALELVDPPGCRNSISSLSI PSTSRGGPVPNSPYSESYYARSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEAR TDRPSQQLRSLNGEWTRPVAAH" primer_bind 1831..1853 /label=M13/pUC Forward /note="In lacZ gene" primer_bind 1845..1862 /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind 1846..1862 /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" primer_bind 1872..1891 /label=T7 /note="T7 promoter, forward primer" promoter 1872..1890 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" misc_feature 1899..2006 /label=MCS /note="pBluescript multiple cloning site" primer_bind 1915..1931 /label=pBluescriptKS /note="For pBluescript vector" primer_bind 1916..1932 /label=KS primer /note="common sequencing primer, one of multiple similar variants" primer_bind complement(1966..1982) /label=SK primer /note="common sequencing primer, one of multiple similar variants" primer_bind complement(1966..1982) /label=pBluescriptSK /note="For pBluescript vector" primer_bind complement(2019..2039) /label=T3 /note="T3 promoter, forward primer" promoter complement(2019..2037) /label=T3 promoter /note="promoter for bacteriophage T3 RNA polymerase" primer_bind complement(2058..2074) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(2058..2074) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(2071..2093) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 2082..2098 /label=lac operator /bound_moiety="lac repressor encoded by lacI" /note="The lac repressor binds to the lac operator to inhibit transcription in E. coli. This inhibition can be relieved by adding lactose or isopropyl-beta-D-thiogalactopyranoside (IPTG)." promoter complement(2106..2136) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 2151..2172 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(2289..2306) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(2460..3048) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(2540..2559) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(3219..4079) /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 3842..3861 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(4080..4184) /gene="bla" /label=AmpR promoter primer_bind 4252..4270 /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" ORIGIN 1 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca 61 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg 121 ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc 181 accatagatc aacgacatta ctatatatat aatataggaa gcatttaata gaacagcatc 241 gtaatatatg tgtactttgc agttatgacg ccagatggca gtagtggaag atattcttta 301 ttgaaaaata gcttgtcacc ttacgtacaa tcttgatccg gagcttttct ttttttgccg 361 attaagaatt aattcggtcg aaaaaagaaa aggagagggc caagagggag ggcattggtg 421 actattgagc acgtgagtat acgtgattaa gcacacaaag gcagcttgga gtatgtctgt 481 tattaatttc acaggtagtt ctggtccatt ggtgaaagtt tgcggcttgc agagcacaga 541 ggccgcagaa tgtgcactag attccgatgc tgacttgctg ggtattatat gtgtgcccaa 601 tagaaagaga acaattgacc cggttattgc aaggaaaatt tcaagtcttg taaaagcata 661 taaaaatagt tcaggcactc cgaaatactt ggttggcgtg tttcgtaatc aacctaagga 721 ggatgttttg gctctggtca atgattacgg cattgatatc gtccaactgc atggagatga 781 gtcgtggcaa gaataccaag agttcctcgg tttgccagtt attaaaagac tcgtatttcc 841 aaaagactgc aacatactac tcagtgcagc ttcacagaaa cctcattcgt ttattccctt 901 gtttgattca gaagcaggtg ggacaggtga acttttggat tggaactcga tttctgactg 961 ggttggaagg caagagagcc ccgagagctt acattttatg ttagctggtg gactgacgcc 1021 agaaaatgtt ggtgatgcgc ttagattaaa tggcgttatt ggtgttgatg taagcggagg 1081 tgtggagaca aatggtgtaa aagactctaa caaaatagca aatttcgtca aaaatgctaa 1141 gaaataggtt attactgagt agtatttatt taagtattgt ttgtgcactt gccgatctat 1201 gcggtgtgaa ataccgcaca gatgcgtaag gagaaaatac cgcatcagga aattgtaagc 1261 gttaatattt tgttaaaatt cgcgttaaat ttttgttaaa tcagctcatt ttttaaccaa 1321 taggccgaaa tcggcaaaat cccttataaa tcaaaagaat agaccgagat agggttgagt 1381 gttgttccag tttggaacaa gagtccacta ttaaagaacg tggactccaa cgtcaaaggg 1441 cgaaaaaccg tctatcaggg cgatggccca ctacgtgaac catcacccta atcaagtttt 1501 ttggggtcga ggtgccgtaa agcactaaat cggaacccta aagggagccc ccgatttaga 1561 gcttgacggg gaaagccggc gaacgtggcg agaaaggaag ggaagaaagc gaaaggagcg 1621 ggcgctaggg cgctggcaag tgtagcggtc acgctgcgcg taaccaccac acccgccgcg 1681 cttaatgcgc cgctacaggg cgcgtccatt cgccattcag gctgcgcaac tgttgggaag 1741 ggcgatcggt gcgggcctct tcgctattac gccagctggc gaaaggggga tgtgctgcaa 1801 ggcgattaag ttgggtaacg ccagggtttt cccagtcacg acgttgtaaa acgacggcca 1861 gtgagcgcgc gtaatacgac tcactatagg gcgaattggg taccgggccc cccctcgagg 1921 tcgacggtat cgataagctt gatatcgaat tcctgcagcc cgggggatcc actagttcta 1981 gagcggccgc caccgcggtg gagctccagc ttttgttccc tttagtgagg gttaattgcg 2041 cgcttggcgt aatcatggtc atagctgttt cctgtgtgaa attgttatcc gctcacaatt 2101 ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta atgagtgagc 2161 taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc 2221 cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct 2281 tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca 2341 gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac 2401 atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt 2461 ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg 2521 cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc 2581 tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc 2641 gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc 2701 aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac 2761 tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt 2821 aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct 2881 aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc 2941 ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt 3001 ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg 3061 atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc 3121 atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa 3181 tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag 3241 gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg 3301 tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga 3361 gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag 3421 cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa 3481 gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc 3541 atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca 3601 aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg 3661 atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat 3721 aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc 3781 aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg 3841 gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg 3901 gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt 3961 gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca 4021 ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata 4081 ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac 4141 atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa 4201 gtgccacctg acgtctaaga aaccattatt atcatgacat taacctataa aaataggcgt 4261 atcacgaggc cctttcgtc //