LOCUS       Exported                6112 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  Yeast episomal vector for expression of His-tagged ubiquitin; URA3 
            marker.
ACCESSION   .
VERSION     .
KEYWORDS    YEp195-CUP1-His-Ubi
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6112)
  AUTHORS   Davies AA, Ulrich HD
  TITLE     Detection of PCNA modifications in Saccharomyces cerevisiae.
  JOURNAL   Methods Mol Biol. 2012;920:543-67. doi: 
            10.1007/978-1-61779-998-3_36.
  PUBMED    22941627
REFERENCE   2  (bases 1 to 6112)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6112
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     promoter        1..235
                     /gene="S. cerevisiae CUP1"
                     /label=CUP1 promoter
                     /note="inducible metallothionein promoter, regulated by 
                     Cu2+"
     CDS             282..302
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=7xHis
                     /translation="HHHHHHH"
     CDS             309..536
                     /codon_start=1
                     /product="Saccharomyces cerevisiae ubiquitin"
                     /label=ubiquitin
                     /translation="MQIFVKTLTGKTITLEVESSDTIDNVKSKIQDKEGIPPDQQRLIF
                     AGKQLEDGRTLSDYNIQKESTLHLVLRLRGG"
     terminator      561..748
                     /gene="S. cerevisiae ADH1"
                     /label=ADH1 terminator
                     /note="transcription terminator for the S. cerevisiae 
                     alcohol dehydrogenase 1 (ADH1) gene"
     primer_bind     complement(616..640)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     primer_bind     complement(772..788)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(772..788)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(785..807)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    796..812
                     /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(820..850)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    865..886
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1003..1020)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1174..1762)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1254..1273)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(1933..2793)
                     /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     2556..2575
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2794..2898)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     2966..2984
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3009..3224
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     CDS             3225..4028
                     /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"
     rep_origin      4139..5485
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     primer_bind     5906..5928
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     5920..5937
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     5921..5937
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 ctagttagaa aaagacattt ttgctgtcag tcactgtcaa gagattcttt tgctggcatt
       61 tcttctagaa gcaaaaagag cgatgcgtct tttccgctga accgttccag caaaaaagac
      121 taccaacgca atatggattg tcagaatcat ataaaagaga agcaaataac tccttgtctt
      181 gtatcaattg cattataata tcttcttgtt agtgcaatat catatagaag tcatcgaaat
      241 agatattaag aaaaacaaac tgtaacggat ccattatggg ccatcatcac catcatcacc
      301 atagcggcat gcaaattttt gtcaagactt taaccggtaa gactattacc ctggaagttg
      361 aatcttctga cactattgac aatgtcaagt ccaagatcca agacaaggaa ggtattccac
      421 ctgaccaaca aagattgatc tttgctggta agcaattgga agatggtaga actttgtccg
      481 actacaacat ccaaaaggaa tctactctac acttggtctt gagattgaga ggtggttgac
      541 tgcagccaag ctaattccgg gcgaatttct tatgatttat gatttttatt attaaataag
      601 ttataaaaaa aataagtgta tacaaatttt aaagtgactc ttaggtttta aaacgaaaat
      661 tcttattctt gagtaactct ttcctgtagg tcaggttgct ttctcaggta tagcatgagg
      721 tcgctcttat tgaccacacc tctaccggca tgcaagcttg gcgtaatcat ggtcatagct
      781 gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat
      841 aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc
      901 actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg
      961 cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct
     1021 gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt
     1081 atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc
     1141 caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga
     1201 gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata
     1261 ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac
     1321 cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg
     1381 taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc
     1441 cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag
     1501 acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt
     1561 aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt
     1621 atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg
     1681 atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac
     1741 gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca
     1801 gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac
     1861 ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac
     1921 ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt
     1981 tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt
     2041 accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt
     2101 atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc
     2161 cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa
     2221 tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg
     2281 tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt
     2341 gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc
     2401 agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt
     2461 aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg
     2521 gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac
     2581 tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc
     2641 gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt
     2701 tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg
     2761 aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat attattgaag
     2821 catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa
     2881 acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgtct aagaaaccat
     2941 tattatcatg acattaacct ataaaaatag gcgtatcacg aggccctttc gtcttcaaga
     3001 attagctttt caattcatca tttttttttt attctttttt ttgatttcgg tttccttgaa
     3061 atttttttga ttcggtaatc tccgaacaga aggaagaacg aaggaaggag cacagactta
     3121 gattggtata tatacgcata tgtagtgttg aagaaacatg aaattgccca gtattcttaa
     3181 cccaactgca cagaacaaaa acatgcagga aacgaagata aatcatgtcg aaagctacat
     3241 ataaggaacg tgctgctact catcctagtc ctgttgctgc caagctattt aatatcatgc
     3301 acgaaaagca aacaaacttg tgtgcttcat tggatgttcg taccaccaag gaattactgg
     3361 agttagttga agcattaggt cccaaaattt gtttactaaa aacacatgtg gatatcttga
     3421 ctgatttttc catggagggc acagttaagc cgctaaaggc attatccgcc aagtacaatt
     3481 ttttactctt cgaagacaga aaatttgctg acattggtaa tacagtcaaa ttgcagtact
     3541 ctgcgggtgt atacagaata gcagaatggg cagacattac gaatgcacac ggtgtggtgg
     3601 gcccaggtat tgttagcggt ttgaagcagg cggcagaaga agtaacaaag gaacctagag
     3661 gccttttgat gttagcagaa ttgtcatgca agggctccct atctactgga gaatatacta
     3721 agggtactgt tgacattgcg aagagcgaca aagattttgt tatcggcttt attgctcaaa
     3781 gagacatggg tggaagagat gaaggttacg attggttgat tatgacaccc ggtgtgggtt
     3841 tagatgacaa gggagacgca ttgggtcaac agtatagaac cgtggatgat gtggtctcta
     3901 caggatctga cattattatt gttggaagag gactatttgc aaagggaagg gatgctaagg
     3961 tagagggtga acgttacaga aaagcaggct gggaagcata tttgagaaga tgcggccagc
     4021 aaaactaaaa aactgtatta taagtaaatg catgtatact aaactcacaa attagagctt
     4081 caatttaatt atatcagtta ttacccaatt ctcatgtttg acagcttatc atcggatcga
     4141 tccaatatca aaggaaatga tagcattgaa ggatgagact aatccaattg aggagtggca
     4201 gcatatagaa cagctaaagg gtagtgctga aggaagcata cgataccccg catggaatgg
     4261 gataatatca caggaggtac tagactacct ttcatcctac ataaatagac gcatataagt
     4321 acgcatttaa gcataaacac gcactatgcc gttcttctca tgtatatata tatacaggca
     4381 acacgcagat ataggtgcga cgtgaacagt gagctgtatg tgcgcagctc gcgttgcatt
     4441 ttcggaagcg ctcgttttcg gaaacgcttt gaagttccta ttccgaagtt cctattctct
     4501 agctagaaag tataggaact tcagagcgct tttgaaaacc aaaagcgctc tgaagacgca
     4561 ctttcaaaaa accaaaaacg caccggactg taacgagcta ctaaaatatt gcgaataccg
     4621 cttccacaaa cattgctcaa aagtatctct ttgctatata tctctgtgct atatccctat
     4681 ataacctacc catccacctt tcgctccttg aacttgcatc taaactcgac ctctacattt
     4741 tttatgttta tctctagtat tactctttag acaaaaaaat tgtagtaaga actattcata
     4801 gagtgaatcg aaaacaatac gaaaatgtaa acatttccta tacgtagtat atagagacaa
     4861 aatagaagaa accgttcata attttctgac caatgaagaa tcatcaacgc tatcactttc
     4921 tgttcacaaa gtatgcgcaa tccacatcgg tatagaatat aatcggggat gcctttatct
     4981 tgaaaaaatg cacccgcagc ttcgctagta atcagtaaac gcgggaagtg gagtcaggct
     5041 ttttttatgg aagagaaaat agacaccaaa gtagccttct tctaacctta acggacctac
     5101 agtgcaaaaa gttatcaaga gactgcatta tagagcgcac aaaggagaaa aaaagtaatc
     5161 taagatgctt tgttagaaaa atagcgctct cgggatgcat ttttgtagaa caaaaaagaa
     5221 gtatagattc tttgttggta aaatagcgct ctcgcgttgc atttctgttc tgtaaaaatg
     5281 cagctcagat tctttgtttg aaaaattagc gctctcgcgt tgcatttttg ttttacaaaa
     5341 atgaagcaca gattcttcgt tggtaaaata gcgctttcgc gttgcatttc tgttctgtaa
     5401 aaatgcagct cagattcttt gtttgaaaaa ttagcgctct cgcgttgcat ttttgttcta
     5461 caaaatgaag cacagatgct tcgttaacaa agatatgcta ttgaagtgca agatggaaac
     5521 gcagaaaatg aaccggggat gcgacgtgca agattaccta tgcaatagat gcaatagttt
     5581 ctccaggaac cgaaatacat acattgtctt ccgtaaagcg ctagactata tattattata
     5641 caggttcaaa tatactatct gtttcaggga aaactcccag gttcggatgt tcaaaattca
     5701 atgatgggta acaagtacga tccgattatg cggtgtgaaa taccgcacag atgcgtaagg
     5761 agaaaatacc gcatcaggcg ccattcgcca ttcaggctgc gcaactgttg ggaagggcga
     5821 tcggtgcggg cctcttcgct attacgccag ctggcgaaag ggggatgtgc tgcaaggcga
     5881 ttaagttggg taacgccagg gttttcccag tcacgacgtt gtaaaacgac ggccagtgaa
     5941 ttcgagctcc cattaccgac atttgggcgc tatacgtgca tatgttcatg tatgtatctg
     6001 tatttaaaac acttttgtat tatttttcct catatatgtg tataggttta tacggatgat
     6061 ttaattatta cttcaccacc ctttatttca ggctgatatc ttagccttgt ta
//
