LOCUS       Exported                7783 bp ds-DNA     circular SYN 15-SEP-2020
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7783)
  AUTHORS   Lindquist Lab
  TITLE     Direct Submission
  JOURNAL   Exported Tuesday, Sep 15, 2020 from SnapGene 5.1.5
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7783
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     promoter        142..172
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    180..196
                     /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     204..220
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     209..262
                     /label=sup_for
     CDS             216..233
                     /codon_start=1
                     /gene="lacZ"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITP"
     misc_feature    233
                     /label=MCS
                     /note="pUC19 multiple cloning site"
     misc_feature    234..733
                     /label=Sup35 promoter
                     /note="Sup35 promoter"
     primer_bind     673..693
                     /label=sup35seq
     primer_bind     complement(702..746)
                     /label=sup_rev
     5'UTR           734..750
                     /note="5'UTR of RPL8A										"
     CDS             751..2283
                     /codon_start=1
                     /label=OLE1
                     /translation="MPTSGTTIELIDDQFPKDDSASSGIVDEVDLTEANILATGLNKKA
                     PRIVNGFGSLMGSKEMVSVEFDKKGNEKKSNLDRLLEKDNQEKEEAKTKIHISEQPWTL
                     NNWHQHLNWLNMVLVCGMPMIGWYFALSGKVPLHLNVFLFSVFYYAVGGVSITAGYHRL
                     WSHRSYSAHWPLRLFYAIFGCASVEGSAKWWGHSHRIHHRYTDTLRDPYDARRGLWYSH
                     MGWMLLKPNPKYKARADITDMTDDWTIRFQHRHYILLMLLTAFVIPTLICGYFFNDYMG
                     GLIYAGFIRVFVIQQATFCINSLAHYIGTQPFDDRRTPRDNWITAIVTFGEGYHNFHHE
                     FPTDYRNAIKWYQYDPTKVIIYLTSLVGLAYDLKKFSQNAIEEALIQQEQKKINKKKAK
                     INWGPVLTDLPMWDKQTFLAKSKENKGLVIISGIVHDVSGYISEHPGGETLIKTALGKD
                     ATKAFSGGVYRHSNAAQNVLADMRVAVIKESKNSAIRMASKRGEIYETGKFF"
     misc_feature    2284..2289
                     /label=MCS
                     /note="color: #a6acb3"
     terminator      2306..2447
                     /label=C Albicans Tadh1
                     /note="This feature has 3 segments: 1: 2770 .. 3130 / 
                     #00ffff 2: 3137 .. 3403 / #00ffff 3: 3410 .. 3464 / 
                     #00ffff"
     misc_feature    2306..2393
                     /label=ADH1 terminator from C.albicans
                     /note="color: #ffffff; direction: RIGHT"
     primer_bind     complement(2416..2472)
                     /label=ADHterm_rev
     misc_feature    2421..2428
                     /label=PacI
                     /note="PacI"
     CDS             2448..2734
                     /codon_start=1
                     /gene="lacZ"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="RVPSSNSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEART
                     DRPSQQLRSLNGEWRLMRYFLLTHLCGISHRIYRWAILMKNILNLLSYY"
     misc_feature    2448..2469
                     /label=MCS
                     /note="pUC19 multiple cloning site"
     primer_bind     complement(2470..2486)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    3244..4407
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN4 centromere fused to the 
                     autonomously replicating sequence ARS1/ARS416"
     CDS             complement(4742..5545)
                     /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"
     promoter        complement(5546..5766)
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     promoter        5877..5981
                     /gene="bla"
                     /label=AmpR promoter
     CDS             5982..6842
                     /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"
     rep_origin      7013..7601
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number colE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctggca
       61 cgacaggttt cccgactgga aagcgggcag tgagcgcaac gcaattaatg tgagttagct
      121 cactcattag gcaccccagg ctttacactt tatgcttccg gctcgtatgt tgtgtggaat
      181 tgtgagcgga taacaatttc acacaggaaa cagctatgac catgattacg ccacaaccac
      241 acaaaaatca tacaacgaat ggtatatgct tcatttcttt gtttcgcatt agctgcgcta
      301 tttgactcaa attattattt tttactaaga cgacgcgtca cagtgttcga gtctgtgtca
      361 tttcttttgt aattctctta aaccacttca taaagttgtg aagttcatag caaaattctt
      421 ccgcaaaaag atgaatctta gttctcagcc caccaaaaga ggtacatgct aagatcatac
      481 agaagttatt gtcacttctt accttgctct taaatgtaca ttacaaccgg gtattatatc
      541 ttacatcatc gtataatatg atctttcttt atggagaaaa tttttttttc actcgaccaa
      601 agctcccatt gcttctgaag agtgtagtgt atattggtac atcttctctt gaaagactcc
      661 attgtactgt aacaaaaagc ggtttcttca tcgacttgct cggaataaca tctatatctg
      721 cccactagca acaaaaacaa ctaattcgaa atgccaactt ctggaactac tattgaattg
      781 attgacgacc aatttccaaa ggatgactct gccagcagtg gcattgtcga cgaagtcgac
      841 ttaacggaag ctaatatttt ggctactggt ttgaataaga aagcaccaag aattgtcaac
      901 ggttttggtt ctttaatggg ctccaaggaa atggtttccg tggaattcga caagaaggga
      961 aacgaaaaga agtccaattt ggatcgtctg ctagaaaagg acaaccaaga aaaagaagaa
     1021 gctaaaacta aaattcacat ctccgaacaa ccatggactt tgaataactg gcaccaacat
     1081 ttgaactggt tgaacatggt tcttgtttgt ggtatgccaa tgattggttg gtactttgct
     1141 ctctctggta aagtgccttt gcatttaaac gttttccttt tctccgtttt ctactacgct
     1201 gtcggtggtg tttctattac tgccggttac catagattat ggtctcacag atcttactcc
     1261 gctcactggc cattgagatt attctacgct atcttcggtt gtgcttccgt tgaagggtcc
     1321 gctaaatggt ggggccactc tcacagaatt caccatcgtt acactgatac cttgagagat
     1381 ccttatgacg ctcgtagagg tctatggtac tcccacatgg gatggatgct tttgaagcca
     1441 aatccaaaat acaaggctag agctgatatt accgatatga ctgatgattg gaccattaga
     1501 ttccaacaca gacactacat cttgttgatg ttgttaaccg ctttcgtcat tccaactctt
     1561 atctgtggtt actttttcaa cgactatatg ggtggtttga tctatgccgg ttttattcgt
     1621 gtctttgtca ttcaacaagc taccttttgc attaactcct tggctcatta catcggtacc
     1681 caaccattcg atgacagaag aacccctcgt gacaactgga ttactgccat tgttactttc
     1741 ggtgaaggtt accataactt ccaccacgaa ttcccaactg attacagaaa cgctattaag
     1801 tggtaccaat acgacccaac taaggttatc atctatttga cttctttagt tggtctagca
     1861 tacgacttga agaaattctc tcaaaatgct attgaagaag ccttgattca acaagaacaa
     1921 aagaagatca ataaaaagaa ggctaagatt aactggggtc cagttttgac tgatttgcca
     1981 atgtgggaca aacaaacctt cttggctaag tctaaggaaa acaagggttt ggttatcatt
     2041 tctggtattg ttcacgacgt atctggttat atctctgaac atccaggtgg tgaaacttta
     2101 attaaaactg cattaggtaa ggacgctacc aaggctttca gtggtggtgt ctaccgtcac
     2161 tcaaatgccg ctcaaaatgt cttggctgat atgagagtgg ctgttatcaa ggaaagtaag
     2221 aactctgcta ttagaatggc tagtaagaga ggtgaaatct acgaaactgg taagttcttt
     2281 taaggccgcc accgcggtgg agctctaagc aaatagctaa attatatacg aattaatatt
     2341 atgattaagt gtttacgtga gtgcgatatt tttattacta tcttatacag ttgtatatac
     2401 tctataaaat gagttgtcta ttaattaacg cgatgaatgc tttctggccc gggtaccgag
     2461 ctcgaattca ctggccgtcg ttttacaacg tcgtgactgg gaaaaccctg gcgttaccca
     2521 acttaatcgc cttgcagcac atcccccttt cgccagctgg cgtaatagcg aagaggcccg
     2581 caccgatcgc ccttcccaac agttgcgcag cctgaatggc gaatggcgcc tgatgcggta
     2641 ttttctcctt acgcatctgt gcggtatttc acaccgcata tatcgctggg ccattctcat
     2701 gaagaatatc ttgaatttat tgtcatatta ctagttggtg tggaagtcct aatatcggtg
     2761 atcaatatag tggttgacat gctggctagt caacattgag ccttttgatc atgcaaatat
     2821 attacggtat tttacaatca aatatcaaac ttaactattg actttataac ttatttaggt
     2881 ggtaacattc ttataaaaaa gaaaaaaatt actgcaaaac agtactagct tttaacttgt
     2941 atcctaggtt atctatgctg tctcaccata gagaatatta cctatttcag aatgtatgtc
     3001 catgattcgc cgggtaaata catataatac acaaatctgg cttaataaag tctataatat
     3061 atctcataaa gaagtgctaa attggctagt gctatatatt tttaagaaaa tttcttttga
     3121 ctaagtccat atcgactttg taaaagttca cattagcata catatattac acgagccaga
     3181 aatagtaact tttgcctaaa tcacaaattg caaaatttaa ttgcttgcaa aaggtcacat
     3241 gcttataatc aactttttta aaaatttaaa atactttttt attttttatt tttaaacata
     3301 aatgaaataa tttatttatt gtttatgatt accgaaacat aaaacctgct caagaaaaag
     3361 aaactgtttt gtccttggaa aaaaagcact acctaggagc ggccaaaatg ccgaggcttt
     3421 catagcttaa actctttaca gaaaataggc attatagatc agttcgagtt ttcttattct
     3481 tccttccggt tttatcgtca cagttttaca gtaaataagt atcacctctt agagttcgat
     3541 gataagctgt caaacatgag aattaattcc cacatgttaa aatagtgaag gagcatgttc
     3601 ggcacacagt ggaccgaacg tggggtaagt gcactagggt ccggttaaac ggatctcgca
     3661 ttgatgaggc aacgctaatt atcaacatat agattgttat ctatctgcat gaacacgaaa
     3721 tctttacttg acgacttgag gctgatggtg tttatgcaaa gaaaccactg tgtttaatat
     3781 gtgtcactgt ttgatattac tgtcagcgta gaagataata gtaaaagcgg ttaataagtg
     3841 tatttgagat aagtgtgata aagtttttac agcgaaaaga cgataaatac aagaaaatga
     3901 ttacgaggat acggagagag gtatgtacat gtgtatttat atactaagct gccggcggtt
     3961 gtttgcaaga ccgagaaaag gctagcaaga atcgggtcat tgtagcgtat gcgcctgtga
     4021 acattctctt caacaagttt gattccattg cggtgaaatg gtaaaagtca accccctgcg
     4081 atgtatattt tcctgtacaa tcaatcaaaa agccaaatga tttagcatta tctttacatc
     4141 ttgttatttt acagatttta tgtttagatc ttttatgctt gcttttcaaa aggcttgcag
     4201 gcaagtgcac aaacaatact taaataaata ctactcagta ataacctatt tcttagcatt
     4261 tttgacgaaa tttgctattt tgttagagtc ttttacacca tttgtctcca cacctccgct
     4321 tacatcaaca ccaataacgc catttaatct aagcgcatca ccaacatttt ctggcgtcag
     4381 tccaccagct aacataaaat gtaagctctc ggggctctct tgccttccaa cccagtcaga
     4441 aatcgagttc caatccaaaa gttcacctgt cccacctgct tctgaatcaa acaagggaat
     4501 aaacgaatga ggtttctgtg aagctgcact gagtagtatg ttgcagtctt ttggaaatac
     4561 gagtctttta ataactggca aaccgaggaa ctcttggtat tcttgccacg actcatctcc
     4621 atgcagttgg acgatcgatg ataagctgtc aaacatgaga attgggtaat aactgatata
     4681 attaaattga agctctaatt tgtgagttta gtatacatgc atttacttat aatacagttt
     4741 tttagttttg ctggccgcat cttctcaaat atgcttccca gcctgctttt ctgtaacgtt
     4801 caccctctac cttagcatcc cttccctttg caaatagtcc tcttccaaca ataataatgt
     4861 cagatcctgt agagaccaca tcatccacgg ttctatactg ttgacccaat gcgtctccct
     4921 tgtcatctaa acccacaccg ggtgtcataa tcaaccaatc gtaaccttca tctcttccac
     4981 ccatgtctct ttgagcaata aagccgataa caaaatcttt gtcgctcttc gcaatgtcaa
     5041 cagtaccctt agtatattct ccagtagata gggagccctt gcatgacaat tctgctaaca
     5101 tcaaaaggcc tctaggttcc tttgttactt cttctgccgc ctgcttcaaa ccgctaacaa
     5161 tacctgggcc caccacaccg tgtgcattcg taatgtctgc ccattctgct attctgtata
     5221 cacccgcaga gtactgcaat ttgactgtat taccaatgtc agcaaatttt ctgtcttcga
     5281 agagtaaaaa attgtacttg gcggataatg cctttagcgg cttaactgtg ccctccatgg
     5341 aaaaatcagt caagatatcc acatgtgttt ttagtaaaca aattttggga cctaatgctt
     5401 caactaactc cagtaattcc ttggtggtac gaacatccaa tgaagcacac aagtttgttt
     5461 gcttttcgtg catgatatta aatagcttgg cagcaacagg actaggatga gtagcagcac
     5521 gttccttata tgtagctttc gacatgattt atcttcgttt cctgcatgtt tttgttctgt
     5581 gcagttgggt taagaatact gggcaatttc atgtttcttc aacactacat atgcgtatat
     5641 ataccaatct aagtctgtgc tccttccttc gttcttcctt ctgttcggag attaccgaat
     5701 caaaaaaatt tcaaagaaac cgaaatcaaa aaaaagaata aaaaaaaaat gatgaattga
     5761 attgaaaagc taattcttga agacgaaagg gcctcgtgat acgcctattt ttataggtta
     5821 atgtcatgat aataatggtt tcttagacgt caggtggcac ttttcgggga aatgtgcgcg
     5881 gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat
     5941 aaccctgata aatgcttcaa taatattgaa aaaggaagag tatgagtatt caacatttcc
     6001 gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct cacccagaaa
     6061 cgctggtgaa agtaaaagat gctgaagatc agttgggtgc acgagtgggt tacatcgaac
     6121 tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgt tttccaatga
     6181 tgagcacttt taaagttctg ctatgtggcg cggtattatc ccgtattgac gccgggcaag
     6241 agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtac tcaccagtca
     6301 cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct gccataacca
     6361 tgagtgataa cactgcggcc aacttacttc tgacaacgat cggaggaccg aaggagctaa
     6421 ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg gaaccggagc
     6481 tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgtagca atggcaacaa
     6541 cgttgcgcaa actattaact ggcgaactac ttactctagc ttcccggcaa caattaatag
     6601 actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt ccggctggct
     6661 ggtttattgc tgataaatct ggagccggtg agcgtgggtc tcgcggtatc attgcagcac
     6721 tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg agtcaggcaa
     6781 ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt aagcattggt
     6841 aactgtcaga ccaagtttac tcatatatac tttagattga tttaaaactt catttttaat
     6901 ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg
     6961 agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc
     7021 ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg
     7081 tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag
     7141 cgcagatacc aaatactgtt cttctagtgt agccgtagtt aggccaccac ttcaagaact
     7201 ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg
     7261 gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc
     7321 ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg
     7381 aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg
     7441 cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag
     7501 ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc
     7561 gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct
     7621 ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct gcgttatccc
     7681 ctgattctgt ggataaccgt attaccgcct ttgagtgagc tgataccgct cgccgcagcc
     7741 gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga aga
//
