LOCUS       Exported                8291 bp ds-DNA     circular SYN 13-MAY-2021
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    piHGpd/G81S
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8291)
  AUTHORS   Nathan DF, Vos MH, Lindquist S
  TITLE     Identification of SSF1, CNS1, and HCH1 as multicopy suppressors of a
            Saccharomyces cerevisiae Hsp90 loss-of-function mutation.
  JOURNAL   Proc Natl Acad Sci U S A 1999 Feb 16;96(4):1409-14.
  PUBMED    9990037
REFERENCE   2  (bases 1 to 8291)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 13, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..8291
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     promoter        1..644
                     /gene="S. cerevisiae TDH3"
                     /label=GAP promoter
                     /note="promoter for glyceraldehyde-3-phosphate 
                     dehydrogenase"
     primer_bind     537..560
                     /label=GPDpro-F
                     /note="S. cerevisiae GPD promoter, forward primer"
     primer_bind     complement(3847..3863)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3847..3863)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     primer_bind     complement(3895..3914)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3896..3914)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3921..3938)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3921..3937)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3930..3952)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      4078..4533
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4165..4184)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4375..4396
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(4793..5455)
                     /codon_start=1
                     /gene="S. cerevisiae HIS3"
                     /product="imidazoleglycerol-phosphate dehydratase, required
                     for histidine biosynthesis"
                     /label=HIS3
                     /note="yeast auxotrophic marker"
                     /translation="MTEQKALVKRITNETKIQIAISLKGGPLAIEHSIFPEKEAEAVAE
                     QATQSQVINVHTGIGFLDHMIHALAKHSGWSLIVECIGDLHIDDHHTTEDCGIALGQAF
                     KEALGAVRGVKRFGSGFAPLDEALSRAVVDLSNRPYAVVELGLQREKVGDLSCEMIPHF
                     LESFAEASRITLHVDCLRGKNDHHRSESAFKALAVAIREATSPNGTNDVPSTKGVLM"
     promoter        complement(5456..5643)
                     /gene="S. cerevisiae HIS3"
                     /label=HIS3 promoter
     primer_bind     complement(5790..5809)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5909..5931
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5969..5987)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6055..6159
                     /gene="bla"
                     /label=AmpR promoter
     CDS             6160..7020
                     /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(6378..6397)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7191..7779
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7680..7699
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7933..7950
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    8067..8088
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        8103..8133
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8141..8157
                     /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     8146..8168
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     8165..8181
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     8165..8181
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        8202..8220
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     8248..8264
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     8249..8265
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 tcattatcaa tactgccatt tcaaagaata cgtaaataat taatagtagt gattttccta
       61 actttattta gtcaaaaaat tagcctttta attctgctgt aacccgtaca tgcccaaaat
      121 agggggcggg ttacacagaa tatataacat cgtaggtgtc tgggtgaaca gtttattcct
      181 ggcatccact aaatataatg gagcccgctt tttaagctgg catccagaaa aaaaaagaat
      241 cccagcacca aaatattgtt ttcttcacca accatcagtt cataggtcca ttctcttagc
      301 gcaactacag agaacagggg cacaaacagg caaaaaacgg gcacaacctc aatggagtga
      361 tgcaacctgc ctggagtaaa tgatgacaca aggcaattga cccacgcatg tatctatctc
      421 attttcttac accttctatt accttctgct ctctctgatt tggaaaaagc tgaaaaaaaa
      481 ggttgaaacc agttccctga aattattccc ctacttgact aataagtata taaagacggt
      541 aggtattgat tgtaattctg taaatctatt tcttaaactt cttaaattct acttttatag
      601 ttagtctttt ttttagtttt aaaacaccaa gaacttagtt tcgacggatc cgattcttaa
      661 catctgtgac ctcctcattt cttcccgctg tattagagtt caagaaatca tacctgatag
      721 aaaatagagt cctataaaca aaagcacaaa caaacacgca aagatatggc tagtgaaact
      781 tttgaatttc aagctgaaat tactcagttg atgagtttga tcatcaacac cgtctattct
      841 aacaaggaaa ttttcttgag agaactgata tctaatgcct cggatgcgtt ggataaaatt
      901 agatacaaat ctttgtctga tccaaagcaa ttggaaacag aaccagatct ctttattaga
      961 atcactccaa agccagagca aaaagttttg gaaatcagag attctagtat tggtatgacc
     1021 aaggctgaat tgattaataa cttgggtacc attgccaagt ctggtaccaa agccttcatg
     1081 gaagctctat ctgctggtgc cgatgtatcc atgattggtc aattcggtgt tggtttttac
     1141 tctttattct tagttgccga cagagttcag gttatttcaa agagcaacga cgacgaacaa
     1201 tacatctggg aatccaacgc tggtggttct ttcactgtta ctctagacga agttaatgaa
     1261 agaattggta ggggtaccat cttgaggtta ttcttgaaag atgaccaatt ggagtacttg
     1321 gaagaaaaga gaataaagga agttatcaag agacattctg agttcgtggc ctacccaatc
     1381 caattagtcg tcaccaagga agttgaaaag gaagttccaa ttccagaaga agaaaagaaa
     1441 gacgaggaaa agaaggatga ggaaaagaag gatgaagacg acaagaaacc aaaattggaa
     1501 gaagtcgatg aagaagagga aaagaagcca aagacgaaaa aagttaaaga agaagttcaa
     1561 gagatagaag aactaaacaa gactaagcct ttgtggacta gaaacccatc tgatatcact
     1621 caagaagaat acaatgcttt ctataagtct atttcaaacg actgggaaga cccattgtac
     1681 gttaagcatt tctccgttga aggtcaattg gaatttagag ctatcttatt cattccaaag
     1741 agagcaccat tcgacttgtt tgagagtaaa aagaagaaga ataatatcaa gttgtacgtt
     1801 cgtcgtgttt tcatcactga tgaagctgaa gacttgattc cagagtggtt atctttcgtc
     1861 aagggtgttg ttgactctga ggatttacca ttgaatttgt ccagagaaat gttacaacaa
     1921 aataagatca tgaaggttat tagaaagaac attgtcaaaa agttgattga agccttcaac
     1981 gaaattgctg aagactctga acaatttgaa aagttctact cggctttctc caaaaatatc
     2041 aagttgggtg tacatgaaga tacccaaaac agggctgctt tggctaagtt gttacgttac
     2101 aactctacca agtccgtaga tgagttgact tccttaactg attacgttac cagaatgcca
     2161 gaacaccaaa agaacatcta ctacatcact ggtgaatctc taaaggctgt cgaaaagtct
     2221 ccatttttgg atgccttgaa ggctaaaaac ttcgaggttt tgttcttgac cgacccaatt
     2281 gatgaatacg ccttcactca attgaaggaa ttcgaaggta aaactttggt tgacattact
     2341 aaagatttcg aattggaaga aactgacgaa gaaaaagctg aaagagagaa ggagatcaaa
     2401 gaatatgaac cattgaccaa ggccttgaaa gaaattttgg gtgaccaagt ggagaaagtt
     2461 gttgtttctt acaaattgtt ggatgcccca gctgctatca gaactggtca atttggttgg
     2521 tctgctaaca tggaaagaat catgaaggct caagccttga gagactcttc catgtcctcc
     2581 tacatgtctt ccaagaagac tttcgaaatt tctccaaaat ctccaattat caaggaattg
     2641 aaaaagagag ttgacgaagg tggtgctcaa gacaagactg tcaaggactt gactaagtta
     2701 ttatatgaaa ctgctttgtt gacttccggc ttcagtttgg acgaaccaac ttcctttgca
     2761 tcaagaatta acagattgat ctctttgggt ttgaacattg atgaggatga agaaacagag
     2821 actgctccag aagcatccac cgcagctccg gttgaagagg ttccagctga caccgaaatg
     2881 gaagaggtag attaggtaga acatcatggc cttgaatagg ttataaacaa aacataatat
     2941 aacgtatagg tattcgaatg aataaataag tatgtaaata gggcatctgc atggaaataa
     3001 ctgggtaaaa cattacaatg tattttttta gagttttcac tattccgacg cgtctagtgt
     3061 gtcatacaaa acgcgaagaa tctgaaatcg tgaaatatac ttaagaatat aactgaaata
     3121 gcttaagcat aaatggactt tactgcgaag ataaagggta tgttgaaact catctgcttt
     3181 cattatgcgt ccaactaatt aacattgcat attttcgtat actatttttt tgtgtagagc
     3241 tcgaacgaga actctcagag acgagtgact ataagacctt gcagaagaag accataagtc
     3301 ttcgaagtga gctgaatact ctgtcacatt ctttgacatc ttatgaaaaa gaacacttct
     3361 ccaatgatat agaaaatgta ttaaaatcta ttaatgcaaa attatctgag tcaaagggca
     3421 aaaaacgact attctccttc aaacagaaaa attcatcatc agccgttcat aagaatgttg
     3481 aacgtacgga gcttgcgaat gcgccggcgt ataccactac tttgaaaaag cattatgtct
     3541 tggagaaggg tgattcagct tttgaaaatt tagagttttg tactgttaca tcaacaacag
     3601 attatagtgg caatagtgcc ctttctggtt ctttgtgctt tcgcaatatt actaaatgtg
     3661 tcataaactt acaaaggatt ttctttcaaa cagggagtat tttcataact gattgtacgg
     3721 attctataat ttttctacgg tctccttccg acaaggattt ccaaatccgt ttacgtgact
     3781 tgaaaaattg caaaattttg atagagaaac tctcgccaag tatcgacctg caggcatgca
     3841 agctcggatc cactagttct agagcggccg ccaccgcggt ggagctccaa ttcgccctat
     3901 agtgagtcgt attacaattc actggccgtc gttttacaac gtcgtgactg ggaaaaccct
     3961 ggcgttaccc aacttaatcg ccttgcagca catccccctt tcgccagctg gcgtaatagc
     4021 gaagaggccc gcaccgatcg cccttcccaa cagttgcgca gcctgaatgg cgaatggacg
     4081 cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta
     4141 cacttgccag cgccctagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt
     4201 tcgccggctt tccccgtcaa gctctaaatc gggggctccc tttagggttc cgatttagtg
     4261 ctttacggca cctcgacccc aaaaaacttg attagggtga tggttcacgt agtgggccat
     4321 cgccctgata gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac
     4381 tcttgttcca aactggaaca acactcaacc ctatctcggt ctattctttt gatttataag
     4441 ggattttgcc gatttcggcc tattggttaa aaaatgagct gatttaacaa aaatttaacg
     4501 cgaattttaa caaaatatta acgcttacaa tttcctgatg cggtattttc tccttacgca
     4561 tctgtgcggt atttcacacc gcatagatcc gtcgagttca agagaaaaaa aaagaaaaag
     4621 caaaaagaaa aaaggaaagc gcgcctcgtt cagaatgaca cgtatagaat gatgcattac
     4681 cttgtcatct tcagtatcat actgttcgta tacatactta ctgacattca taggtataca
     4741 tatatacaca tgtatatata tcgtatgctg cagctttaaa taatcggtgt cactacataa
     4801 gaacaccttt ggtggaggga acatcgttgg taccattggg cgaggtggct tctcttatgg
     4861 caaccgcaag agccttgaac gcactctcac tacggtgatg atcattcttg cctcgcagac
     4921 aatcaacgtg gagggtaatt ctgctagcct ctgcaaagct ttcaagaaaa tgcgggatca
     4981 tctcgcaaga gagatctcct actttctccc tttgcaaacc aagttcgaca actgcgtacg
     5041 gcctgttcga aagatctacc accgctctgg aaagtgcctc atccaaaggc gcaaatcctg
     5101 atccaaacct ttttactcca cgcacggccc ctagggcctc tttaaaagct tgaccgagag
     5161 caatcccgca gtcttcagtg gtgtgatggt cgtctatgtg taagtcacca atgcactcaa
     5221 cgattagcga ccagccggaa tgcttggcca gagcatgtat catatggtcc agaaacccta
     5281 tacctgtgtg gacgttaatc acttgcgatt gtgtggcctg ttctgctact gcttctgcct
     5341 ctttttctgg gaagatcgag tgctctatcg ctaggggacc accctttaaa gagatcgcaa
     5401 tctgaatctt ggtttcattt gtaatacgct ttactagggc tttctgctct gtcatctttg
     5461 ccttcgttta tcttgcctgc tcatttttta gtatattctt cgaagaaatc acattacttt
     5521 atataatgta taattcatta tgtgataatg ccaatcgcta agaaaaaaaa agagtcatcc
     5581 gctaggtgga aaaaaaaaaa tgaaaatcat taccgaggca taaaaaaata tagagtgtac
     5641 tagaggaggc caagagtaat agaaaaagaa aattgcggga aaggactgtg ttatgacttc
     5701 cctgactaat gccgtgttca aacgatacct ggcagtgact cctagcgctc accaagctct
     5761 taaaacggga atttatggtg cactctcagt acaatctgct ctgatgccgc atagttaagc
     5821 cagccccgac acccgccaac acccgctgac gcgccctgac gggcttgtct gctcccggca
     5881 tccgcttaca gacaagctgt gaccgtctcc gggagctgca tgtgtcagag gttttcaccg
     5941 tcatcaccga aacgcgcgag acgaaagggc ctcgtgatac gcctattttt ataggttaat
     6001 gtcatgataa taatggtttc ttagacgtca ggtggcactt ttcggggaaa tgtgcgcgga
     6061 acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa
     6121 ccctgataaa tgcttcaata atattgaaaa aggaagagta tgagtattca acatttccgt
     6181 gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg
     6241 ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac gagtgggtta catcgaactg
     6301 gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgttt tccaatgatg
     6361 agcactttta aagttctgct atgtggcgcg gtattatccc gtattgacgc cgggcaagag
     6421 caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtactc accagtcaca
     6481 gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc cataaccatg
     6541 agtgataaca ctgcggccaa cttacttctg acaacgatcg gaggaccgaa ggagctaacc
     6601 gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga accggagctg
     6661 aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg
     6721 ttgcgcaaac tattaactgg cgaactactt actctagctt cccggcaaca attaatagac
     6781 tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc ggctggctgg
     6841 tttattgctg ataaatctgg agccggtgag cgtgggtctc gcggtatcat tgcagcactg
     6901 gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag tcaggcaact
     6961 atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggtaa
     7021 ctgtcagacc aagtttactc atatatactt tagattgatt taaaacttca tttttaattt
     7081 aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag
     7141 ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct
     7201 ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt
     7261 tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg
     7321 cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt caagaactct
     7381 gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc
     7441 gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg
     7501 tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa
     7561 ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg
     7621 gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg
     7681 ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga
     7741 tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt
     7801 ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct
     7861 gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg ccgcagccga
     7921 acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcccaat acgcaaaccg
     7981 cctctccccg cgcgttggcc gattcattaa tgcagctggc acgacaggtt tcccgactgg
     8041 aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc tcactcatta ggcaccccag
     8101 gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa ttgtgagcgg ataacaattt
     8161 cacacaggaa acagctatga ccatgattac gccaagctcg aaattaaccc tcactaaagg
     8221 gaacaaaagc tggtaccggg ccccccctcg aggtcgacgg tatcgataag cttggctgca
     8281 ggtcgagttt a
//
