LOCUS       Exported                8438 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    pGALSc104(WT)
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8438)
  AUTHORS   Schirmer EC, Homann OR, Kowal AS, Lindquist S
  TITLE     Dominant gain-of-function mutations in Hsp104p reveal crucial roles 
            for the middle region.
  JOURNAL   Mol Biol Cell 2004 May;15(5):2061-72. Epub 2004 Feb 20.
  PUBMED    14978213
REFERENCE   2  (bases 1 to 8438)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..8438
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     promoter        27..691
                     /gene="GAL1 and GAL10 genes of S. cerevisiae"
                     /label=GAL1,10 promoter
                     /note="divergent inducible promoter, regulated by Gal4"
     protein_bind    243..360
                     /label=UAS
                     /bound_moiety="Gal4"
                     /note="upstream activating sequence mediating 
                     Gal4-dependent induction"
     primer_bind     655..678
                     /label=GAL1
                     /note="S. cerevisiae GAL1 promoter, forward primer"
     primer_bind     complement(3553..3572)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3554..3572)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3579..3596)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3579..3595)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3588..3610)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      3740..4195
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3827..3846)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4037..4058
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(4455..5114)
                     /codon_start=1
                     /gene="S. cerevisiae HIS3"
                     /product="imidazoleglycerol-phosphate dehydratase, required
                     for histidine biosynthesis"
                     /label=HIS3
                     /note="yeast auxotrophic marker"
                     /translation="MTEQKALVKRITNETKIQIAISLKGGPLAIEHSIFPEKEAEAVAE
                     QATQSQVINVHTGIGFLDHMIHALAKHSGWSLIVECIGDLHIDDHHTTEDCGIALGQAF
                     KEALLARGVKRFGSGFAPLDEALSRAVVDLSNRPYAVVELGLQREKVGDLSCEMIPHFL
                     ESFAEASRITLHVDCLRGKNDHHRSESAFKALAVAIREATSPNGTNDVPSTKGVLM"
     promoter        complement(5115..5301)
                     /gene="S. cerevisiae HIS3"
                     /label=HIS3 promoter
     primer_bind     complement(5448..5467)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5567..5589
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5627..5645)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     misc_feature    5687..6190
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     promoter        6227..6331
                     /gene="bla"
                     /label=AmpR promoter
     CDS             6332..7192
                     /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     complement(6550..6569)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7363..7951
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7852..7871
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     8105..8122
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        8275..8305
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8313..8329
                     /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)."
     primer_bind     8318..8340
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     8337..8353
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     8337..8353
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        8374..8392
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     8421..8437
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     8422..8438
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 cgataagctt gatatcgaat tcgaattttc aaaaattctt actttttttt tggatggacg
       61 caaagaagtt taataatcat attacatggc attaccacca tatacatatc catatacata
      121 tccatatcta atcttactta tatgttgtgg aaatgtaaag agccccatta tcttagccta
      181 aaaaaacctt ctctttggaa ctttcagtaa tacgcttaac tgctcattgc tatattgaag
      241 tacggattag aagccgccga gcgggtgaca gccctccgaa ggaagactct cctccgtgcg
      301 tcctcgtctt caccggtcgc gttcctgaaa cgcagatgtg cctcgcgccg cactgctccg
      361 aacaataaag attctacaat actagctttt atggttatga agaggaaaaa ttggcagtaa
      421 cctggcccca caaaccttca aatgaacgaa tcaaattaac aaccatagga tgataatgcg
      481 attagttttt tagccttatt tctggggtaa ttaatcagcg aagcgatgat ttttgatcta
      541 ttaacagata tataaatgca aaaactgcat aaccacttta actaatactt tcaacatttt
      601 cggtttgtat tacttcttat tcaaatgtaa taaaagtatc aacaaaaaat tgttaatata
      661 cctctatact ttaacgtcaa ggagaaaaaa cccggatccg ggatgaacga ccaaacgcaa
      721 tttacagaaa gggctctaac gattttgacg ttggctcaaa aattggcttc ggatcatcaa
      781 catccacaat tacaacctat acatattcta gctgccttca ttgaaacgcc agaagatgga
      841 tcagtccctt acctacagaa tctaattgag aagggccgtt acgactatga tcttttcaag
      901 aaagtggtta atagaaatct agtaagaatt cctcaacagc aacctgcacc tgcggagata
      961 actccaagtt atgctttggg gaaagtcctt caagacgctg ctaagattca aaaacaacag
     1021 aaggactcat ttatagcgca agaccatata ttgtttgctc tattcaatga ttcgtctatt
     1081 cagcaaatat ttaaggaagc tcaagtagat attgaggcca tcaagcaaca agctcttgaa
     1141 cttcgtggta acactagaat tgactctcgt ggcgctgata cgaacacacc tttggaatat
     1201 ttatcaaagt acgccattga tatgactgag caggctcgtc aaggtaaact tgaccctgtc
     1261 atcggccgtg aagaagaaat aagaagcact attagagttt tagcaagaag aattaagtcc
     1321 aacccatgtt taattggtga gccaggtatc ggtaagaccg ctattattga aggtgttgct
     1381 caaagaatca ttgacgatga cgttcccact atcttacaag gcgctaaatt gttcagtcta
     1441 gatttggccg cattaaccgc aggtgctaaa tacaaaggtg atttcgaaga aagattcaaa
     1501 ggtgttttga aggaaatcga agaatcaaag actctaattg tgttattcat tgatgaaatt
     1561 cacatgttaa tgggtaatgg taaggacgac gctgctaaca tcttgaagcc agctttgtcc
     1621 agaggccaat tgaaggtcat cggtgccacc accaataacg aatatagatc tattgtggaa
     1681 aaggatggtg cctttgaaag aagattccag aaaattgaag tcgctgaacc aagtgtgaga
     1741 caaacagtgg ccatattgag aggtctgcaa ccaaagtatg aaatacatca tggtgtaagg
     1801 attctggata gcgccttagt caccgcggct caattagcca agcgttactt gccatataga
     1861 agattgccag attctgcttt ggatttagtt gatatttctt gtgctggtgt cgccgtcgca
     1921 agagattcta agccagaaga attggattcc aaggaacgtc aattgcaatt gattcaagta
     1981 gagataaaag ctctagagag agatgaagat gccgactcca ccactaaaga tagattaaag
     2041 ttagctaggc agaaggaagc ttcattgcaa gaagaattgg aacctctaag acaacgttac
     2101 aatgaagaaa agcatggcca tgaagaattg acacaagcta aaaagaaatt ggatgaactg
     2161 gaaaacaagg cccttgatgc tgaacgtaga tatgatactg ctaccgccgc tgatttaagg
     2221 tacttcgcca tcccagatat caaaaagcaa atcgaaaagc ttgaagatca ggttgctgag
     2281 gaagagagac gtgctggtgc caactccatg atccaaaatg tggtcgactc agacaccatt
     2341 tctgaaacag ctgcaagatt gactggtatc cctgttaaga agttgtcaga atctgaaaat
     2401 gaaaaattga ttcatatgga acgtgactta tcatctgaag tcgtgggcca aatggatgcc
     2461 attaaagctg tttccaatgc cgttagattg tctagatcag gtttagctaa tccaaggcaa
     2521 ccagcatcct tcttattttt aggtttgtcc ggttccggta aaactgaatt ggctaaaaaa
     2581 gttgctggat ttttgtttaa tgatgaggac atgatgatca gggtcgattg ttctgaatta
     2641 agcgagaagt atgcggtctc taagttgttg ggtaccacgg caggttatgt cgggtacgat
     2701 gaaggtggct ttttaactaa ccaactgcaa tacaaaccat actccgtttt gttattcgat
     2761 gaagtagaaa aggcacatcc tgatgttttg actgtcatgc tacaaatgtt ggatgacggt
     2821 agaattacta gtggtcaagg taagacgatc gactgttcca attgtattgt catcatgact
     2881 tccaatctag gtgctgaatt tatcaattct caacaaggat caaagatcca agaatctacc
     2941 aagaatttgg tcatgggtgc tgttaggcaa catttcagac cagaattttt gaacagaatt
     3001 tctagtatag tcattttcaa caagctatct agaaaagcta ttcataagat cgtggatatt
     3061 cgtttgaagg aaattgaaga gagattcgag caaaatgata aacattacaa gttgaattta
     3121 actcaagagg ccaaggactt cttggccaaa tatggttatt ccgatgatat gggtgcacgt
     3181 ccactgaaca ggttaattca aaacgaaatt ttgaacaaac tggcactaag gatcttaaag
     3241 aatgaaatca aggataagga aactgtcaat gtcgtcttga agaagggtaa atctcgtgat
     3301 gaaaatgttc ctgaggaagc tgaagaatgt ctggaagttc taccaaatca cgaagctact
     3361 ataggggctg acacgttagg tgatgacgat aatgaggaca gtatggaaat tgatgatgac
     3421 ctagattaat ttaatatagt gtgattttta aaaactttcg aacaagaatc agtaatataa
     3481 tatatataat taataaaact aatggaattt gtttaattga acttgacacc cgagcaaaag
     3541 agctccaatt cgccctatag tgagtcgtat tacaattcac tggccgtcgt tttacaacgt
     3601 cgtgactggg aaaaccctgg cgttacccaa cttaatcgcc ttgcagcaca tccccctttc
     3661 gccagctggc gtaatagcga agaggcccgc accgatcgcc cttcccaaca gttgcgcagc
     3721 ctgaatggcg aatggcgcga cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg
     3781 gttacgcgca gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc tttcgctttc
     3841 ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcgggggctc
     3901 cctttagggt tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgattagggt
     3961 gatggttcac gtagtgggcc atcgccctga tagacggttt ttcgcccttt gacgttggag
     4021 tccacgttct ttaatagtgg actcttgttc caaactggaa caacactcaa ccctatctcg
     4081 gtctattctt ttgatttata agggattttg ccgatttcgg cctattggtt aaaaaatgag
     4141 ctgatttaac aaaaatttaa cgcgaatttt aacaaaatat taacgtttac aatttcctga
     4201 tgcggtattt tctccttacg catctgtgcg gtatttcaca ccgcatagat ccgtcgagtt
     4261 caagagaaaa aaaaagaaaa agcaaaaaga aaaaaggaaa gcgcgcctcg ttcagaatga
     4321 cacgtataga atgatgcatt accttgtcat cttcagtatc atactgttcg tatacatact
     4381 tactgacatt cataggtata catatataca catgtatata tatcgtatgc tgcagcttta
     4441 aataatcggt gtcactacat aagaacacct ttggtggagg gaacatcgtt ggtaccattg
     4501 ggcgaggtgg cttctcttat ggcaaccgca agagccttga acgcactctc actacggtga
     4561 tgatcattct tgcctcgcag acaatcaacg tggagggtaa ttctgctagc ctctgcaaag
     4621 ctttcaagaa aatgcgggat catctcgcaa gagagatctc ctactttctc cctttgcaaa
     4681 ccaagttcga caactgcgta cggcctgttc gaaagatcta ccaccgctct ggaaagtgcc
     4741 tcatccaaag gcgcaaatcc tgatccaaac ctttttactc cacgcgccag tagggcctct
     4801 ttaaaagctt gaccgagagc aatcccgcag tcttcagtgg tgtgatggtc gtctatgtgt
     4861 aagtcaccaa tgcactcaac gattagcgac cagccggaat gcttggccag agcatgtatc
     4921 atatggtcca gaaaccctat acctgtgtgg acgttaatca cttgcgattg tgtggcctgt
     4981 tctgctactg cttctgcctc tttttctggg aagatcgagt gctctatcgc taggggacca
     5041 ccctttaaag agatcgcaat ctgaatcttg gtttcatttg taatacgctt tactagggct
     5101 ttctgctctg tcatctttgc cttcgtttat cttgcctgct cattttttag tatattcttc
     5161 gaagaaatca cattacttta tataatgtat aattcattat gtgataatgc caatcgctaa
     5221 gaaaaaaaaa gagtcatccg ctaggggaaa aaaaaaaatg aaaatcatta ccgaggcata
     5281 aaaaaatata gagtgtacta gaggaggcca agagtaatag aaaaagaaaa ttgcgggaaa
     5341 ggactgtgtt atgacttccc tgactaatgc cgtgttcaaa cgatacctgg cagtgactcc
     5401 tagcgctcac caagctctta aaacgggaat ttatggtgca ctctcagtac aatctgctct
     5461 gatgccgcat agttaagcca gccccgacac ccgccaacac ccgctgacgc gccctgacgg
     5521 gcttgtctgc tcccggcatc cgcttacaga caagctgtga ccgtctccgg gagctgcatg
     5581 tgtcagaggt tttcaccgtc atcaccgaaa cgcgcgagac gaaagggcct cgtgatacgc
     5641 ctatttttat aggttaatgt catgataata atggtttctt aggacggatc gcttgcctgt
     5701 aacttacacg cgcctcgtat cttttaatga tggaataatt tgggaattta ctctgtgttt
     5761 atttattttt atgttttgta tttggatttt agaaagtaaa taaagaaggt agaagagtta
     5821 cggaatgaag aaaaaaaaat aaacaaaggt ttaaaaaatt tcaacaaaaa gcgtacttta
     5881 catatatatt tattagacaa gaaaagcaga ttaaatagat atacattcga ttaacgataa
     5941 gtaaaatgta aaatcacagg attttcgtgt gtggtcttct acacagacaa gatgaaacaa
     6001 ttcggcatta atacctgaga gcaggaagag caagataaaa ggtagtattt gttggcgatc
     6061 cccctagagt cttttacatc ttcggaaaac aaaaactatt ttttctttaa tttctttttt
     6121 tactttctat ttttaattta tatatttata ttaaaaaatt taaattataa ttatttttat
     6181 agcacgtgat gaaaaggacc caggtggcac ttttcgggga aatgtgcgcg gaacccctat
     6241 ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata
     6301 aatgcttcaa taatattgaa aaaggaagag tatgagtatt caacatttcc gtgtcgccct
     6361 tattcccttt tttgcggcat tttgccttcc tgtttttgct cacccagaaa cgctggtgaa
     6421 agtaaaagat gctgaagatc agttgggtgc acgagtgggt tacatcgaac tggatctcaa
     6481 cagcggtaag atccttgaga gttttcgccc cgaagaacgt tttccaatga tgagcacttt
     6541 taaagttctg ctatgtggcg cggtattatc ccgtattgac gccgggcaag agcaactcgg
     6601 tcgccgcata cactattctc agaatgactt ggttgagtac tcaccagtca cagaaaagca
     6661 tcttacggat ggcatgacag taagagaatt atgcagtgct gccataacca tgagtgataa
     6721 cactgcggcc aacttacttc tgacaacgat cggaggaccg aaggagctaa ccgctttttt
     6781 gcacaacatg ggggatcatg taactcgcct tgatcgttgg gaaccggagc tgaatgaagc
     6841 cataccaaac gacgagcgtg acaccacgat gcctgtagca atggcaacaa cgttgcgcaa
     6901 actattaact ggcgaactac ttactctagc ttcccggcaa caattaatag actggatgga
     6961 ggcggataaa gttgcaggac cacttctgcg ctcggccctt ccggctggct ggtttattgc
     7021 tgataaatct ggagccggtg agcgtgggtc tcgcggtatc attgcagcac tggggccaga
     7081 tggtaagccc tcccgtatcg tagttatcta cacgacgggg agtcaggcaa ctatggatga
     7141 acgaaataga cagatcgctg agataggtgc ctcactgatt aagcattggt aactgtcaga
     7201 ccaagtttac tcatatatac tttagattga tttaaaactt catttttaat ttaaaaggat
     7261 ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt
     7321 ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct
     7381 gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc
     7441 ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc
     7501 aaatactgtc cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc
     7561 gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc
     7621 gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg
     7681 aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata
     7741 cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta
     7801 tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc
     7861 ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg
     7921 atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt
     7981 cctggccttt tgctggcctt ttgctcacat gttctttcct gcgttatccc ctgattctgt
     8041 ggataaccgt attaccgcct ttgagtgagc tgataccgct cgccgcagcc gaacgaccga
     8101 gcgcagcgag tcagtgagcg aggaagcgga agagcgccca atacgcaaac cgcctctccc
     8161 cgcgcgttgg ccgattcatt aatgcagctg gcacgacagg tttcccgact ggaaagcggg
     8221 cagtgagcgc aacgcaatta atgtgagtta cctcactcat taggcacccc aggctttaca
     8281 ctttatgctt ccggctcgta tgttgtgtgg aattgtgagc ggataacaat ttcacacagg
     8341 aaacagctat gaccatgatt acgccaagct cggaattaac cctcactaaa gggaacaaaa
     8401 gctgggtacc gggccccccc tcgaggtcga cggtatcg
//
