LOCUS Exported 6835 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS pRSII325 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6835) 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 6835) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..6835 /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" CDS complement(664..1758) /codon_start=1 /gene="S. cerevisiae LEU2" /product="3-isopropylmalate dehydrogenase, required for leucine biosynthesis" /label=LEU2 /note="yeast auxotrophic marker" /translation="MSAPKKIVVLPGDHVGQEITAEAIKVLKAISDVRSNVKFDFENHL IGGAAIDATGVPLPDEALEASKKADAVLLGAVGGPKWGTGSVRPEQGLLKIRKELQLYA NLRPCNFASDSLLDLSPIKPQFAKGTDFVVVRELVGGIYFGKRKEDDGDGVAWDSEQYT VPEVQRITRMAAFMALQHEPPLPIWSLDKANVLASSRLWRKTVEETIKNEFPTLKVQHQ LIDSAAMILVKNPTHLNGIIITSNMFGDIISDEASVIPGSLGLLPSASLASLPDKNTAF GLYEPCHGSAPDLPKNKVNPIATILSAAMMLKLSLNLPEEGKAIEDAVKKVLDAGIRTG DLGGSNSTTEVGDAVAEEVKKILA" promoter complement(1759..2166) /gene="S. cerevisiae LEU2" /label=LEU2 promoter rep_origin complement(2467..2922) /direction=LEFT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(2604..2625) /label=F1ori-F /note="F1 origin, forward primer" primer_bind 2816..2835 /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 3048..3070 /label=M13/pUC Forward /note="In lacZ gene" primer_bind 3062..3079 /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind 3063..3079 /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" primer_bind 3086..3105 /label=T7 /note="T7 promoter, forward primer" promoter 3086..3104 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" misc_feature 3113..3220 /label=MCS /note="pBluescript multiple cloning site" primer_bind 3137..3153 /label=SK primer /note="common sequencing primer, one of multiple similar variants" primer_bind 3137..3153 /label=pBluescriptSK /note="For pBluescript vector" primer_bind complement(3187..3203) /label=KS primer /note="common sequencing primer, one of multiple similar variants" primer_bind complement(3188..3204) /label=pBluescriptKS /note="For pBluescript vector" promoter complement(3232..3250) /label=T3 promoter /note="promoter for bacteriophage T3 RNA polymerase" primer_bind complement(3271..3287) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(3271..3287) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(3284..3306) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 3295..3311 /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(3319..3349) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 3364..3385 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(3502..3519) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(3673..4261) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(3753..3772) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(4432..5292) /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 5055..5074 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(5293..5397) /gene="bla" /label=AmpR promoter primer_bind 6808..6826 /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 accatatcga ctacgtcgtt aaggccgttt ctgacagagt aaaattcttg agggaacttt 241 caccattatg ggaaatggtt caagaaggta ttgacttaaa ctccatcaaa tggtcaggtc 301 attgagtgtt ttttatttgt tgtatttttt tttttttaga gaaaatcctc caatatataa 361 attaggaatc atagtttcat gattttctgt tacacctaac tttttgtgtg gtgccctcct 421 ccttgtcaat attaatgtta aagtgcaatt ctttttcctt atcacgttga gccattagta 481 tcaatttgct tacctgtatt cctttacatc ctcctttttc tccttcttga taaatgtatg 541 tagattgcgt atatagtttc gtctacccta tgaacatatt ccattttgta atttcgtgtc 601 gtttctatta tgaatttcat ttataaagtt tatgtacaaa tatcataaaa aaagagaatc 661 tttttaagca aggattttct taacttcttc ggcgacagca tcaccgactt cggtggtact 721 gttggaacca cctaaatcac cagttctgat acctgcatcc aaaacctttt taactgcatc 781 ttcaatggcc ttaccttctt caggcaagtt caatgacaat ttcaacatca ttgcagcaga 841 caagatagtg gcgatagggt tgaccttatt ctttggcaaa tctggagcag aaccgtggca 901 tggttcgtac aaaccaaatg cggtgttctt gtctggcaaa gaggccaagg acgcagatgg 961 caacaaaccc aaggaacctg ggataacgga ggcttcatcg gagatgatat caccaaacat 1021 gttgctggtg attataatac catttaggtg ggttgggttc ttaactagga tcatggcggc 1081 agaatcaatc aattgatgtt gaaccttcaa tgtaggaaat tcgttcttga tggtttcctc 1141 cacagttttt ctccataatc ttgaagaggc caaaacatta gctttatcca aggaccaaat 1201 aggcaatggt ggctcatgtt gtagggccat gaaagcggcc attcttgtga ttctttgcac 1261 ttctggaacg gtgtattgtt cactatccca agcgacacca tcaccatcgt cttcctttct 1321 cttaccaaag taaatacctc ccactaattc tctgacaaca acgaagtcag tacctttagc 1381 aaattgtggc ttgattggag ataagtctaa aagagagtcg gatgcaaagt tacatggtct 1441 taagttggcg tacaattgaa gttctttacg gatttttagt aaaccttgtt caggtctaac 1501 actacctgta ccccatttag gaccacccac agcacctaac aaaacggcat cagccttctt 1561 ggaggcttcc agcgcctcat ctggaagtgg aacacctgta gcatctatag cagcaccacc 1621 aattaaatga ttttcgaaat cgaacttgac attggaacga acatcagaaa tagctttaag 1681 aaccttaatg gcttcggctg tgatttcttg accaacgtgg tcacctggca aaacgacgat 1741 cttcttaggg gcagacatta gaatggtata tccttgaaat atatatatat atattgctga 1801 aatgtaaaag gtaagaaaag ttagaaagta agacgattgc taaccaccta ttggaaaaaa 1861 caataggtcc ttaaataata ttgtcaactt caagtattgt gatgcaagca tttagtcatg 1921 aacgcttctc tattctatat gaaaagccgg ttccggcgct ctcacctttc ctttttctcc 1981 caatttttca gttgaaaaag gtatatgcgt caggcgacct ctgaaattaa caaaaaattt 2041 ccagtcatcg aatttgattc tgtgcgatag cgcccctgtg tgttctcgtt atgttgagga 2101 aaaaaataat ggttgctaag agattcgaac tcttgcatct tacgatacct gagtattccc 2161 acagttaact gcggtcaaga tatttcttga atcaggcgcc ttagaccgct cggccaaaca 2221 accaattact tgttgagaaa tagagtataa ttatcctata aatataacgt ttttgaacac 2281 acatgaacaa ggaagtacag gacaattgat tttgaagaga atgtggattt tgatgtaatt 2341 gttgggattc catttttaat aaggcaataa tattaggtat gtagatatac tagaagttct 2401 cctcgaccgg tcgatatgcg gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc 2461 atcaggaaat tgtaagcgtt aatattttgt taaaattcgc gttaaatttt tgttaaatca 2521 gctcattttt taaccaatag gccgaaatcg gcaaaatccc ttataaatca aaagaataga 2581 ccgagatagg gttgagtgtt gttccagttt ggaacaagag tccactatta aagaacgtgg 2641 actccaacgt caaagggcga aaaaccgtct atcagggcga tggcccacta cgtgaaccat 2701 caccctaatc aagttttttg gggtcgaggt gccgtaaagc actaaatcgg aaccctaaag 2761 ggagcccccg atttagagct tgacggggaa agccggcgaa cgtggcgaga aaggaaggga 2821 agaaagcgaa aggagcgggc gctagggcgc tggcaagtgt agcggtcacg ctgcgcgtaa 2881 ccaccacacc cgccgcgctt aatgcgccgc tacagggcgc gtccattcgc cattcaggct 2941 gcgcaactgt tgggaagggc gatcggtgcg ggcctcttcg ctattacgcc agctggcgaa 3001 agggggatgt gctgcaaggc gattaagttg ggtaacgcca gggttttccc agtcacgacg 3061 ttgtaaaacg acggccagtg aattgtaata cgactcacta tagggcgaat tggagctcca 3121 ccgcggtggc ggccgctcta gaactagtgg atcccccggg ctgcaggaat tcgatatcaa 3181 gcttatcgat accgtcgacc tcgagggggg gcccggtacc agcttttgtt ccctttagtg 3241 agggttaatt tcgagcttgg cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta 3301 tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag cctggggtgc 3361 ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt tccagtcggg 3421 aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg 3481 tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg 3541 gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa 3601 cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc 3661 gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc 3721 aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag 3781 ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct 3841 cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta 3901 ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc 3961 cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc 4021 agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt 4081 gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct 4141 gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc 4201 tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca 4261 agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta 4321 agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa 4381 atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg 4441 cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg 4501 actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc 4561 aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc 4621 cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa 4681 ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc 4741 cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg 4801 ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc 4861 cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat 4921 ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg 4981 tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc 5041 ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg 5101 aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat 5161 gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg 5221 gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg 5281 ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct 5341 catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac 5401 atttccccga aaagtgccac ctgacgagca tctgtgcttc attttgtaga acaaaaatgc 5461 aacgcgagag cgctaatttt tcaaacaaag aatctgagct gcatttttac agaacagaaa 5521 tgcaacgcga aagcgctatt ttaccaacga agaatctgtg cttcattttt gtaaaacaaa 5581 aatgcaacgc gagagcgcta atttttcaaa caaagaatct gagctgcatt tttacagaac 5641 agaaatgcaa cgcgagagcg ctattttacc aacaaagaat ctatacttct tttttgttct 5701 acaaaaatgc atcccgagag cgctattttt ctaacaaagc atcttagatt actttttttc 5761 tcctttgtgc gctctataat gcagtctctt gataactttt tgcactgtag gtccgttaag 5821 gttagaagaa ggctactttg gtgtctattt tctcttccat aaaaaaagcc tgactccact 5881 tcccgcgttt actgattact agcgaagctg cgggtgcatt ttttcaagat aaaggcatcc 5941 ccgattatat tctataccga tgtggattgc gcatactttg tgaacagaaa gtgatagcgt 6001 tgatgattct tcattggtca gaaaattatg aacggtttct tctattttgt ctctatatac 6061 tacgtatagg aaatgtttac attttcgtat tgttttcgat tcactctatg aatagttctt 6121 actacaattt ttttgtctaa agagtaatac tagagataaa cataaaaaat gtagaggtcg 6181 agtttagatg caagttcaag gagcgaaagg tggatgggta ggttatatag ggatatagca 6241 cagagatata tagcaaagag atacttttga gcaatgtttg tggaagcggt attcgcaata 6301 ttttagtagc tcgttacagt ccggtgcgtt tttggttttt tgaaagtgcg tcttcagagc 6361 gcttttggtt ttcaaaagcg ctctgaagtt cctatacttt ctagctagag aataggaact 6421 tcggaatagg aacttcaaag cgtttccgaa aacgagcgct tccgaaaatg caacgcgagc 6481 tgcgcacata cagctcactg ttcacgtcgc acctatatct gcgtgttgcc tgtatatata 6541 tatacatgag aagaacggca tagtgcgtgt ttatgcttaa atgcgtactt atatgcgtct 6601 atttatgtag gatgaaaggt agtctagtac ctcctgtgat attatcccat tccatgcggg 6661 gtatcgtatg cttccttcag cactaccctt tagctgttct atatgctgcc actcctcaat 6721 tggattagtc tcatccttca atgctatcat ttcctttgat attggatcat ctaagaaacc 6781 attattatca tgacattaac ctataaaaat aggcgtatca cgaggccctt tcgtc //