LOCUS       sequence-191764-        6616 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Yeast episomal vector for expression of His-tagged ubiquitin; LEU2 
            marker.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6616)
  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 6616)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Methods Mol
            Biol."; date: "2012"; volume: "920:543-67. doi"; pages: " 
            10.1007/978-1-61779-998-3_36"
COMMENT     Sequence Label: YEp181-CUP1-His-Ubi
FEATURES             Location/Qualifiers
     source          1..6616
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     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(join(820..826,827..844,845..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(join(1933..2724,2725..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        3004..3409
                     /gene="S. cerevisiae LEU2"
                     /label=LEU2 promoter
     CDS             3410..4504
                     /codon_start=1
                     /gene="S. cerevisiae LEU2"
                     /product="3-isopropylmalate dehydrogenase, required for 
                     leucine biosynthesis"
                     /label=LEU2
                     /note="yeast auxotrophic marker"
                     /translation="MSAPKKIVVLPGDHVGQEITAEAIKVLKAISDVRSNVKFDFENHL
                     IGGAAIDATGVPLPDEALEASKKADAVLLGAVGGPKWGTGSVRPEQGLLKIRKELQLYA
                     NLRPCNFASDSLLDLSPIKPQFAKGTDFVVVRELVGGIYFGKRKEDDGDGVAWDSEQYT
                     VPEVQRITRMAAFMALQHEPPLPIWSLDKANVLASSRLWRKTVEETIKNEFPTLKVQHQ
                     LIDSAAMILVKNPTHLNGIIITSNMFGDIISDEASVIPGSLGLLPSASLASLPDKNTAF
                     GLYEPCHGSAPDLPKNKVNPIATILSAAMMLKLSLNLPEEGKAIEDAVKKVLDAGIRTG
                     DLGGSNSTTEVGDAVAEEVKKILA"
     rep_origin      4642..5988
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     primer_bind     6410..6432
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     6424..6441
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     6425..6441
                     /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 attaactgtg ggaatactca ggtatcgtaa gatgcaagag ttcgaatctc ttagcaacca
     3061 ttattttttt cctcaacata acgagaacac acaggggcgc tatcgcacag aatcaaattc
     3121 gatgactgga aattttttgt taatttcaga ggtcgcctga cgcatatacc tttttcaact
     3181 gaaaaattgg gagaaaaagg aaaggtgaga gcgccggaac cggcttttca tatagaatag
     3241 agaagcgttc atgactaaat gcttgcatca caatacttga agttgacaat attatttaag
     3301 gacctattgt tttttccaat aggtggttag caatcgtctt actttctaac ttttcttacc
     3361 ttttacattt cagcaatata tatatatatt tcaaggatat accattctaa tgtctgcccc
     3421 taagaagatc gtcgttttgc caggtgacca cgttggtcaa gaaatcacag ccgaagccat
     3481 taaggttctt aaagctattt ctgatgttcg ttccaatgtc aagttcgatt tcgaaaatca
     3541 tttaattggt ggtgctgcta tcgatgctac aggtgttcca cttccagatg aggcgctgga
     3601 agcctccaag aaggctgatg ccgttttgtt aggtgctgtg ggtggtccta aatggggtac
     3661 aggtagtgtt agacctgaac aaggtttact aaaaatccgt aaagaacttc aattgtacgc
     3721 caacttaaga ccatgtaact ttgcatccga ctctctttta gacttatctc caatcaagcc
     3781 acaatttgct aaaggtactg acttcgttgt tgtcagagaa ttagtgggag gtatttactt
     3841 tggtaagaga aaggaagacg atggtgatgg tgtcgcttgg gatagtgaac aatacaccgt
     3901 tccagaagtg caaagaatca caagaatggc cgctttcatg gccctacaac atgagccacc
     3961 attgcctatt tggtccttgg ataaagctaa tgttttggcc tcttcaagat tatggagaaa
     4021 aactgtggag gaaaccatca agaacgaatt tcctacattg aaggttcaac atcaattgat
     4081 tgattctgcc gccatgatcc tagttaagaa cccaacccac ctaaatggta ttataatcac
     4141 cagcaacatg tttggtgata tcatctccga tgaagcctcc gttatcccag gttccttggg
     4201 tttgttgcca tctgcgtcct tggcctcttt gccagacaag aacaccgcat ttggtttgta
     4261 cgaaccatgc cacggttctg ctccagattt gccaaagaat aaggtcaacc ctatcgccac
     4321 tatcttgtct gctgcaatga tgttgaaatt gtcattgaac ttgcctgaag aaggtaaggc
     4381 cattgaagat gcagttaaaa aggttttgga tgcaggtatc agaactggtg atttaggtgg
     4441 ttccaacagt accaccgaag tcggtgatgc tgtcgccgaa gaagttaaga aaatccttgc
     4501 ttaaaaagat tctctttttt tatgatattt gtacataaac tttataaatg aaattcataa
     4561 tagaaacgac acgaaattac aaaatggaat atgttcatag ggtagaatta attctcatgt
     4621 ttgacagctt atcatcggat cgatccaata tcaaaggaaa tgatagcatt gaaggatgag
     4681 actaatccaa ttgaggagtg gcagcatata gaacagctaa agggtagtgc tgaaggaagc
     4741 atacgatacc ccgcatggaa tgggataata tcacaggagg tactagacta cctttcatcc
     4801 tacataaata gacgcatata agtacgcatt taagcataaa cacgcactat gccgttcttc
     4861 tcatgtatat atatatacag gcaacacgca gatataggtg cgacgtgaac agtgagctgt
     4921 atgtgcgcag ctcgcgttgc attttcggaa gcgctcgttt tcggaaacgc tttgaagttc
     4981 ctattccgaa gttcctattc tctagctaga aagtatagga acttcagagc gcttttgaaa
     5041 accaaaagcg ctctgaagac gcactttcaa aaaaccaaaa acgcaccgga ctgtaacgag
     5101 ctactaaaat attgcgaata ccgcttccac aaacattgct caaaagtatc tctttgctat
     5161 atatctctgt gctatatccc tatataacct acccatccac ctttcgctcc ttgaacttgc
     5221 atctaaactc gacctctaca ttttttatgt ttatctctag tattactctt tagacaaaaa
     5281 aattgtagta agaactattc atagagtgaa tcgaaaacaa tacgaaaatg taaacatttc
     5341 ctatacgtag tatatagaga caaaatagaa gaaaccgttc ataattttct gaccaatgaa
     5401 gaatcatcaa cgctatcact ttctgttcac aaagtatgcg caatccacat cggtatagaa
     5461 tataatcggg gatgccttta tcttgaaaaa atgcacccgc agcttcgcta gtaatcagta
     5521 aacgcgggaa gtggagtcag gcttttttta tggaagagaa aatagacacc aaagtagcct
     5581 tcttctaacc ttaacggacc tacagtgcaa aaagttatca agagactgca ttatagagcg
     5641 cacaaaggag aaaaaaagta atctaagatg ctttgttaga aaaatagcgc tctcgggatg
     5701 catttttgta gaacaaaaaa gaagtataga ttctttgttg gtaaaatagc gctctcgcgt
     5761 tgcatttctg ttctgtaaaa atgcagctca gattctttgt ttgaaaaatt agcgctctcg
     5821 cgttgcattt ttgttttaca aaaatgaagc acagattctt cgttggtaaa atagcgcttt
     5881 cgcgttgcat ttctgttctg taaaaatgca gctcagattc tttgtttgaa aaattagcgc
     5941 tctcgcgttg catttttgtt ctacaaaatg aagcacagat gcttcgttaa caaagatatg
     6001 ctattgaagt gcaagatgga aacgcagaaa atgaaccggg gatgcgacgt gcaagattac
     6061 ctatgcaata gatgcaatag tttctccagg aaccgaaata catacattgt cttccgtaaa
     6121 gcgctagact atatattatt atacaggttc aaatatacta tctgtttcag ggaaaactcc
     6181 caggttcgga tgttcaaaat tcaatgatgg gtaacaagta cgatccgata aatgcggtgt
     6241 gaaataccgc acagatgcgt aaggagaaaa taccgcatca ggcgccattc gccattcagg
     6301 ctgcgcaact gttgggaagg gcgatcggtg cgggcctctt cgctattacg ccagctggcg
     6361 aaagggggat gtgctgcaag gcgattaagt tgggtaacgc cagggttttc ccagtcacga
     6421 cgttgtaaaa cgacggccag tgaattcgag ctcccattac cgacatttgg gcgctatacg
     6481 tgcatatgtt catgtatgta tctgtattta aaacactttt gtattatttt tcctcatata
     6541 tgtgtatagg tttatacgga tgatttaatt attacttcac caccctttat ttcaggctga
     6601 tatcttagcc ttgtta
//
