LOCUS Exported 8945 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS 5538 pUG2-SSA1 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 8945) AUTHORS Glover JR, Lindquist S TITLE Hsp104, Hsp70, and Hsp40: a novel chaperone system that rescues previously aggregated proteins. JOURNAL Cell 1998 Jul 10;94(1):73-82. PUBMED 9674429 REFERENCE 2 (bases 1 to 8945) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..8945 /organism="synthetic DNA construct" /mol_type="other DNA" primer_bind complement(2958..2974) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(2958..2974) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(2971..2993) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 2982..2998 /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(3006..3036) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 3051..3072 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(3189..3206) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(3360..3948) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(3440..3459) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(4119..4979) /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 4742..4761 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(4980..5084) /gene="bla" /label=AmpR promoter primer_bind 5152..5170 /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" rep_origin 5528..6870 /label=2u ori /note="yeast 2u plasmid origin of replication" protein_bind 5860..5907 /label=FRT /bound_moiety="FLP recombinase from the Saccharomyces cerevisiae 2u plasmid" /note="FLP-mediated recombination occurs in the 8-bp core sequence TCTAGAAA (Turan and Bode, 2011)." promoter 7462..7677 /gene="S. cerevisiae URA3" /label=URA3 promoter CDS 7678..8481 /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" ORIGIN 1 atgtcaaaag ctgtcggtat tgatttaggt acaacatact cgtgtgttgc tcactttgct 61 aatgatcgtg tggacattat tgccaacgat caaggtaaca gaaccactcc atcttttgtc 121 gctttcactg acactgaaag attgattggt gatgctgcta agaatcaagc tgctatgaat 181 ccttcgaata ccgttttcga cgctaagcgt ttgatcggta gaaacttcaa cgacccagaa 241 gtgcaggctg acatgaagca cttcccattc aagttgatcg atgttgacgg taagccacaa 301 attcaagttg aatttaaggg tgaaaccaag aactttaccc cagaacaaat ctcctccatg 361 gtcttgggta agatgaagga aactgccgaa tcttacttgg gtgccaaggt caatgacgct 421 gtcgtcactg tcccagctta cttcaacgat tctcaaagac aagctaccaa ggatgctggt 481 accattgctg gtttgaatgt cttgcgtatt attaacgaac ctaccgccgc tgccattgct 541 tacggtttgg acaagaaggg taaggaagaa cacgtcttga ttttcgactt gggtggtggt 601 actttcgatg tctctttgtt gtccattgaa gacggtatct ttgaagttaa ggccaccgct 661 ggtgacaccc atttgggtgg tgaagatttt gacaacagat tggtcaacca cttcatccaa 721 gaattcaaga gaaagaacaa gaaggacttg tctaccaacc aaagagcttt gagaagatta 781 agaaccgctt gtgaaagagc caagagaact ttgtcttcct ccgctcaaac ttccgttgaa 841 attgactctt tgttcgaagg tatcgatttc tacacttcca tcaccagagc cagattcgaa 901 gaattgtgtg ctgacttgtt cagatctact ttggacccag ttgaaaaggt cttgagagat 961 gctaaattgg acaaatctca agtcgatgaa attgtcttgg tcggtggttc taccagaatt 1021 ccaaaggtcc aaaaattggt cactgactac ttcaacggta aggaaccaaa cagatctatc 1081 aacccagatg aagctgttgc ttacggtgct gctgttcaag ctgctatttt gactggtgac 1141 gaatcttcca agactcaaga tctattgttg ttggatgtcg ctccattatc cttgggtatt 1201 gaaactgctg gtggtgtcat gaccaagttg attccaagaa actctaccat tccaacaaag 1261 aagtccgaga tcttttccac ttatgctgat aaccaaccag gtgtcttgat tcaagtcttt 1321 gaaggtgaaa gagccaagac taaggacaac aacttgttgg gtaagttcga attgagtggt 1381 attccaccag ctccaagagg tgtcccacaa attgaagtca ctttcgatgt cgactctaac 1441 ggtattttga atgtttccgc cgtcgaaaag ggtactggta agtctaacaa gatcactatt 1501 accaacgaca agggtagatt gtccaaggaa gatatcgaaa agatggttgc tgaagccgaa 1561 aaattcaagg aagaagatga aaaggaatct caaagaattg cttccaagaa ccaattggaa 1621 tccattgctt actctttgaa gaacaccatt tctgaagctg gtgacaaatt ggaacaagct 1681 gacaaggaca ccgtcaccaa gaaggctgaa gagactattt cttggttaga cagcaacacc 1741 actgccagca aggaagaatt cgatgacaag ttgaaggagt tgcaagacat tgccaaccca 1801 atcatgtcta agttgtacca agctggtggt gctccaggtg gcgctgcagg tggtgctcca 1861 ggcggtttcc caggtggtgc tcctccagct ccagaggctg aaggtccaac cgttgaagaa 1921 gttgattaag ccaattggtg cggcaattga taataacgaa aatgtctttt aatgatctgg 1981 gtataatgag gaattttccg aacgttttta ctttatatat atatatacat gtaacatata 2041 ttctatacgc tatagagaaa ggaaattttt caattaaaaa aaaaatagag aaagagtttc 2101 acttcttgat tatcgctaac actaatggtt gaagtactgc tactttaatt ttatagatag 2161 gcaaaaaaaa attattcggg gcgagctggg aattgaaccc agggcctctc gcatgcacca 2221 ttccttgcgg cggcggtgct caacggcctc aacctactac tgggctgctt cctaatgcag 2281 gagtcgcata agggagagcg tcgaccgatg cccttgagag ccttcaaccc agtcagctcc 2341 ttccggtggg cgcggggcat gactatcgtc gccgcactta tgactgtctt ctttatcatg 2401 caactcgtag gacaggtgcc ggcagcgctc tgggtcattt tcggcgagga ccgctttcgc 2461 tggagcgcga cgatgatcgg cctgtcgctt gcggtattcg gaatcttgca cgccctcgct 2521 caagccttcg tcactggtcc cgccaccaaa cgtttcggcg agaagcaggc cattatcgcc 2581 ggcatggcgg ccgacgcgct gggctacgtc ttgctggcgt tcgcgacgcg aggctggatg 2641 gccttcccca ttatgattct tctcgcttcc ggcggcatcg ggatgcccgc gttgcaggcc 2701 atgctgtcca ggcaggtaga tgacgaccat cagggacagc ttcaaggatc gctcgcggct 2761 cttaccagcc taacttcgat cattggaccg ctgatcgtca cggcgattta tgccgcctcg 2821 gcgagcacat ggaacgggtt ggcatggatt gtaggcgccg ccctatacct tgtctgcctc 2881 cccgcgttgc gtcgcggtgc atggagccgg gccacctact gaagtggatt tctttaagag 2941 ctcgaattcg taatcatgtc atagctgttt cctgtgtgaa attgttatcc gctcacaatt 3001 ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta atgagtgagc 3061 taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc 3121 cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct 3181 tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca 3241 gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac 3301 atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt 3361 ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg 3421 cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc 3481 tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc 3541 gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc 3601 aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac 3661 tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt 3721 aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct 3781 aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc 3841 ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt 3901 ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg 3961 atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc 4021 atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa 4081 tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag 4141 gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg 4201 tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga 4261 gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag 4321 cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa 4381 gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc 4441 atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca 4501 aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg 4561 atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat 4621 aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc 4681 aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg 4741 gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg 4801 gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt 4861 gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca 4921 ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata 4981 ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac 5041 atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa 5101 gtgccacctg acgtctaaga aaccattatt atcatgacat taacctataa aaataggcgt 5161 atcacgaggc cctttcgtct tcaagaattc tgaaccagtc ctaaaacgag taaataggac 5221 cggcaattct tcaagcaata aacaggaata ccaattatta aaagataact tagtcagatc 5281 gtacaataaa gctttgaaga aaaatgcgcc ttattcaatc tttgctataa aaaatggccc 5341 aaaatctcac attggaagac atttgatgac ctcatttctt tcaatgaagg gcctaacgga 5401 gttgactaat gttgtgggaa attggagcga taagcgtgct tctgccgtgg ccaggacaac 5461 gtatactcat cagataacag caatacctga tcactacttc gcactagttt ctcggtacta 5521 tgcatatgat ccaatatcaa aggaaatgat agcattgaag gatgagacta atccaattga 5581 ggagtggcag catatagaac agctaaaggg tagtgctgaa ggaagcatac gataccccgc 5641 atggaatggg ataatatcac aggaggtact agactacctt tcatcctaca taaatagacg 5701 catataagta cgcatttaag cataaacacg cactatgccg ttcttctcat gtatatatat 5761 atacaggcaa cacgcagata taggtgcgac gtgaacagtg agctgtatgt gcgcagctcg 5821 cgttgcattt tcggaagcgc tcgttttcgg aaacgctttg aagttcctat tccgaagttc 5881 ctattctcta gaaagtatag gaacttcaga gcgcttttga aaaccaaaag cgctctgaag 5941 acgcactttc aaaaaaccaa aaacgcaccg gactgtaacg agctactaaa atattgcgaa 6001 taccgcttcc acaaacattg ctcaaaagta tctctttgct atatatctct gtgctatatc 6061 cctatataac ctacccatcc acctttcgct ccttgaactt gcatctaaac tcgacctcta 6121 cattttttat gtttatctct agtattactc tttagacaaa aaaattgtag taagaactat 6181 tcatagagtg aatcgaaaac aatacgaaaa tgtaaacatt tcctatacgt agtatataga 6241 gacaaaatag aagaaaccgt tcataatttt ctgaccaatg aagaatcatc aacgctatca 6301 ctttctgttc acaaagtatg cgcaatccac atcggtatag aatataatcg gggatgcctt 6361 tatcttgaaa aaatgcaccc gcagcttcgc tagtaatcag taaacgcggg aagtggagtc 6421 aggctttttt tatggaagag aaaatagaca ccaaagtagc cttcttctaa ccttaacgga 6481 cctacagtgc aaaaagttat caagagactg cattatagag cgcacaaagg agaaaaaaag 6541 taatctaaga tgctttgtta gaaaaatagc gctctcggga tgcatttttg tagaacaaaa 6601 aagaagtata gattctttgt tggtaaaata gcgctctcgc gttgcatttc tgttctgtaa 6661 aaatgcagct cagattcttt gtttgaaaaa ttagcgctct cgcgttgcat ttttgtttta 6721 caaaaatgaa gcacagattc ttcgttggta aaatagcgct ttcgcgttgc atttctgttc 6781 tgtaaaaatg cagctcagat tctttgtttg aaaaattagc gctctcgcgt tgcatttttg 6841 ttctacaaaa tgaagcacag atgcttcgtt aacaaagata tgctattgaa gtgcaagatg 6901 gaaacgcaga aaatgaaccg gggatgcgac gtgcaagatt acctatgcaa tagatgcaat 6961 agtttctcca ggaaccgaaa tacatacatt gtcttccgta aagcgctaga ctatatatta 7021 ttatacaggt tcaaatatac tatctgtttc agggaaaact cccaggttcg gatgttcaaa 7081 attcaatgat gggtaacaag tacgatcgta aatctgtaaa acagtttgtc ggatattagg 7141 ctgtatctcc tcaaagcgta ttcgaatatc attgagaagc tgcagcgtca catcggataa 7201 taatgatggc agccattgta gaagtgcctt ttgcatttct agtctctttc tcggtctagc 7261 tagttttact acatcgcgaa gatagaatct tagatcacac tgcctttgct gagctggatc 7321 aatagagtaa caaaagagtg gtaaggcctc gttaaaggac aaggacctga gcggaagtgt 7381 atcgtacagt agacggagta tactagtata gtctatagtc cgtggaattc tcatgtttga 7441 cagcttatca tcgataagct tttcaattca tcattttttt tttattcttt tttttgattt 7501 cggtttcctt gaaatttttt tgattcggta atctccgaac agaaggaaga acgaaggaag 7561 gagcacagac ttagattggt atatatacgc atatgtagtg ttgaagaaac atgaaattgc 7621 ccagtattct taacccaact gcacagaaca aaaacctgca ggaaacgaag ataaatcatg 7681 tcgaaagcta catataagga acgtgctgct actcatccta gtcctgttgc tgccaagcta 7741 tttaatatca tgcacgaaaa gcaaacaaac ttgtgtgctt cattggatgt tcgtaccacc 7801 aaggaattac tggagttagt tgaagcatta ggtcccaaaa tttgtttact aaaaacacat 7861 gtggatatct tgactgattt ttccatggag ggcacagtta agccgctaaa ggcattatcc 7921 gccaagtaca attttttact cttcgaagac agaaaatttg ctgacattgg taatacagtc 7981 aaattgcagt actctgcggg tgtatacaga atagcagaat gggcagacat tacgaatgca 8041 cacggtgtgg tgggcccagg tattgttagc ggtttgaagc aggcggcaga agaagtaaca 8101 aaggaaccta gaggcctttt gatgttagca gaattgtcat gcaagggctc cctatctact 8161 ggagaatata ctaagggtac tgttgacatt gcgaagagcg acaaagattt tgttatcggc 8221 tttattgctc aaagagacat gggtggaaga gatgaaggtt acgattggtt gattatgaca 8281 cccggtgtgg gtttagatga caagggagac gcattgggtc aacagtatag aaccgtggat 8341 gatgtggtct ctacaggatc tgacattatt attgttggaa gaggactatt tgcaaaggga 8401 agggatgcta aggtagaggg tgaacgttac agaaaagcag gctgggaagc atatttgaga 8461 agatgcggcc agcaaaacta aaaaactgta ttataagtaa atgcatgtat actaaactca 8521 caaattagag cttcaattta attatatcag ttattaccct cgacctcgac tgtacaggat 8581 gttctagcta ctcgagtagc tagaacatcc tgtacagtcg agtagctaga acatcctgta 8641 cagtcgagca gatccgccag gcgtgtatat agcgtggatg gccaggcaac tttagtgctg 8701 acacatacag gcatatatat atgtgtgcga cgacacatga tcatatggca tgcatgtgct 8761 ctgtatgtat ataaaactct tgttttcttc ttttctctaa atattctttc cttatacatt 8821 aggtcctttg tagcataaat tactatactt ctatagacac gcaaacacgg atccaagctt 8881 ggcacatcaa aagaaaagta atcaagtatt acaagaaaca aaaattcaag taaataacag 8941 ataat //