LOCUS Exported 5718 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS pRSII426 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 5718) 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 5718) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..5718 /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 194..409 /gene="S. cerevisiae URA3" /label=URA3 promoter CDS 410..1213 /codon_start=1 /gene="S. cerevisiae URA3" /product="orotidine-5'-phosphate decarboxylase, required for uracil biosynthesis" /label=URA3 /note="yeast auxotrophic marker, counterselectable with 5-fluoroorotic acid (5-FOA)" /translation="MSKATYKERAATHPSPVAAKLFNIMHEKQTNLCASLDVRTTKELL ELVEALGPKICLLKTHVDILTDFSMEGTVKPLKALSAKYNFLLFEDRKFADIGNTVKLQ YSAGVYRIAEWADITNAHGVVGPGIVSGLKQAAEEVTKEPRGLLMLAELSCKGSLSTGE YTKGTVDIAKSDKDFVIGFIAQRDMGGRDEGYDWLIMTPGVGLDDKGDALGQQYRTVDD VVSTGSDIIIVGRGLFAKGRDAKVEGERYRKAGWEAYLRRCGQQN" rep_origin complement(1344..1799) /direction=LEFT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(1481..1502) /label=F1ori-F /note="F1 origin, forward primer" primer_bind 1693..1712 /label=F1ori-R /note="F1 origin, reverse primer" CDS complement(1776..2156) /codon_start=1 /gene="lacZ fragment" /product="LacZ-alpha fragment of beta-galactosidase" /label=lacZ-alpha /translation="MTMITPSAQLTLTKGNKSWSSTAVAAALELVDPPGCRNSISSLSI PSTSRGGPVPNSPYSESYYARSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEAR TDRPSQQLRSLNGEWTRPVAAH" primer_bind 1925..1947 /label=M13/pUC Forward /note="In lacZ gene" primer_bind 1939..1956 /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind 1940..1956 /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" primer_bind 1966..1985 /label=T7 /note="T7 promoter, forward primer" promoter 1966..1984 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" misc_feature 1993..2100 /label=MCS /note="pBluescript multiple cloning site" primer_bind 2009..2025 /label=pBluescriptKS /note="For pBluescript vector" primer_bind 2010..2026 /label=KS primer /note="common sequencing primer, one of multiple similar variants" primer_bind complement(2060..2076) /label=SK primer /note="common sequencing primer, one of multiple similar variants" primer_bind complement(2060..2076) /label=pBluescriptSK /note="For pBluescript vector" primer_bind complement(2113..2133) /label=T3 /note="T3 promoter, forward primer" promoter complement(2113..2131) /label=T3 promoter /note="promoter for bacteriophage T3 RNA polymerase" primer_bind complement(2152..2168) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(2152..2168) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(2165..2187) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 2176..2192 /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(2200..2230) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 2245..2266 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(2383..2400) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(2554..3142) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(2634..2653) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(3313..4173) /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 3936..3955 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(4174..4278) /gene="bla" /label=AmpR promoter rep_origin 4305..5651 /label=2u ori /note="yeast 2u plasmid origin of replication" primer_bind 5691..5709 /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 accataccac cttttcaatt catcattttt tttttattct tttttttgat ttcggtttcc 241 ttgaaatttt tttgattcgg taatctccga acagaaggaa gaacgaagga aggagcacag 301 acttagattg gtatatatac gcatatgtag tgttgaagaa acatgaaatt gcccagtatt 361 cttaacccaa ctgcacagaa caaaaacatg caggaaacga agataaatca tgtcgaaagc 421 tacatataag gaacgtgctg ctactcatcc tagtcctgtt gctgccaagc tatttaatat 481 catgcacgaa aagcaaacaa acttgtgtgc ttcattggat gttcgtacca ccaaggaatt 541 actggagtta gttgaagcat taggtcccaa aatttgttta ctaaaaacac atgtggatat 601 cttgactgat ttttccatgg agggcacagt taagccgcta aaggcattat ccgccaagta 661 caatttttta ctcttcgaag acagaaaatt tgctgacatt ggtaatacag tcaaattgca 721 gtactctgcg ggtgtataca gaatagcaga atgggcagac attacgaatg cacacggtgt 781 ggtgggacca ggtattgtta gcggtttgaa gcaggcggca gaagaagtaa caaaggaacc 841 tagaggcctt ttgatgttag cagaattgtc atgcaagggc tccctatcta ctggagaata 901 tactaagggt actgttgaca ttgcgaagag cgacaaagat tttgttatcg gctttattgc 961 tcaaagagac atgggtggaa gagatgaagg ttacgattgg ttgattatga cacccggtgt 1021 gggtttagat gacaagggag acgcattggg tcaacagtat agaaccgtgg atgatgtggt 1081 ctctacagga tctgacatta ttattgttgg aagaggacta tttgcaaagg gaagggatgc 1141 taaggtagag ggtgaacgtt acagaaaagc aggctgggaa gcatatttga gaagatgcgg 1201 ccagcaaaac taaaaaactg tattataagt aaatgcatgt atactaaact cacaaattag 1261 agcttcaatt taattatatc agttattacc ctatgcggtg tgaaataccg cacagatgcg 1321 taaggagaaa ataccgcatc aggaaattgt aagcgttaat attttgttaa aattcgcgtt 1381 aaatttttgt taaatcagct cattttttaa ccaataggcc gaaatcggca aaatccctta 1441 taaatcaaaa gaatagaccg agatagggtt gagtgttgtt ccagtttgga acaagagtcc 1501 actattaaag aacgtggact ccaacgtcaa agggcgaaaa accgtctatc agggcgatgg 1561 cccactacgt gaaccatcac cctaatcaag ttttttgggg tcgaggtgcc gtaaagcact 1621 aaatcggaac cctaaaggga gcccccgatt tagagcttga cggggaaagc cggcgaacgt 1681 ggcgagaaag gaagggaaga aagcgaaagg agcgggcgct agggcgctgg caagtgtagc 1741 ggtcacgctg cgcgtaacca ccacacccgc cgcgcttaat gcgccgctac agggcgcgtc 1801 cattcgccat tcaggctgcg caactgttgg gaagggcgat cggtgcgggc ctcttcgcta 1861 ttacgccagc tggcgaaagg gggatgtgct gcaaggcgat taagttgggt aacgccaggg 1921 ttttcccagt cacgacgttg taaaacgacg gccagtgagc gcgcgtaata cgactcacta 1981 tagggcgaat tgggtaccgg gccccccctc gaggtcgacg gtatcgataa gcttgatatc 2041 gaattcctgc agcccggggg atccactagt tctagagcgg ccgccaccgc ggtggagctc 2101 cagcttttgt tccctttagt gagggttaat tgcgcgcttg gcgtaatcat ggtcatagct 2161 gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat 2221 aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc 2281 actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg 2341 cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct 2401 gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt 2461 atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc 2521 caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga 2581 gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata 2641 ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac 2701 cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg 2761 taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc 2821 cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag 2881 acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt 2941 aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt 3001 atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg 3061 atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac 3121 gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca 3181 gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac 3241 ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac 3301 ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt 3361 tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt 3421 accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt 3481 atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc 3541 cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa 3601 tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg 3661 tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt 3721 gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc 3781 agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt 3841 aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg 3901 gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac 3961 tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc 4021 gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt 4081 tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg 4141 aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat attattgaag 4201 catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa 4261 acaaataggg gttccgcgca catttccccg aaaagtgcca cctgaacgaa gcatctgtgc 4321 ttcattttgt agaacaaaaa tgcaacgcga gagcgctaat ttttcaaaca aagaatctga 4381 gctgcatttt tacagaacag aaatgcaacg cgaaagcgct attttaccaa cgaagaatct 4441 gtgcttcatt tttgtaaaac aaaaatgcaa cgcgagagcg ctaatttttc aaacaaagaa 4501 tctgagctgc atttttacag aacagaaatg caacgcgaga gcgctatttt accaacaaag 4561 aatctatact tcttttttgt tctacaaaaa tgcatcccga gagcgctatt tttctaacaa 4621 agcatcttag attacttttt ttctcctttg tgcgctctat aatgcagtct cttgataact 4681 ttttgcactg taggtccgtt aaggttagaa gaaggctact ttggtgtcta ttttctcttc 4741 cataaaaaaa gcctgactcc acttcccgcg tttactgatt actagcgaag ctgcgggtgc 4801 attttttcaa gataaaggca tccccgatta tattctatac cgatgtggat tgcgcatact 4861 ttgtgaacag aaagtgatag cgttgatgat tcttcattgg tcagaaaatt atgaacggtt 4921 tcttctattt tgtctctata tactacgtat aggaaatgtt tacattttcg tattgttttc 4981 gattcactct atgaatagtt cttactacaa tttttttgtc taaagagtaa tactagagat 5041 aaacataaaa aatgtagagg tcgagtttag atgcaagttc aaggagcgaa aggtggatgg 5101 gtaggttata tagggatata gcacagagat atatagcaaa gagatacttt tgagcaatgt 5161 ttgtggaagc ggtattcgca atattttagt agctcgttac agtccggtgc gtttttggtt 5221 ttttgaaagt gcgtcttcag agcgcttttg gttttcaaaa gcgctctgaa gttcctatac 5281 tttctagcta gagaatagga acttcggaat aggaacttca aagcgtttcc gaaaacgagc 5341 gcttccgaaa atgcaacgcg agctgcgcac atacagctca ctgttcacgt cgcacctata 5401 tctgcgtgtt gcctgtatat atatatacat gagaagaacg gcatagtgcg tgtttatgct 5461 taaatgcgta cttatatgcg tctatttatg taggatgaaa ggtagtctag tacctcctgt 5521 gatattatcc cattccatgc ggggtatcgt atgcttcctt cagcactacc ctttagctgt 5581 tctatatgct gccactcctc aattggatta gtctcatcct tcaatgctat catttccttt 5641 gatattggat catctaagaa accattatta tcatgacatt aacctataaa aataggcgta 5701 tcacgaggcc ctttcgtc //