LOCUS       sequence-202576-        6022 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express yEGFP-ERG9 ER targeting peptide-CLN2PEST under the control 
            of TEF1 promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6022)
  AUTHORS   Peng B, Plan MR, Chrysanthopoulos P, Hodson MP, Nielsen LK, Vickers 
            CE
  TITLE     A squalene synthase protein degradation method for improved 
            sesquiterpene production in Saccharomyces cerevisiae.
  JOURNAL   Metab Eng. 2017 Jan;39:209-219. doi: 10.1016/j.ymben.2016.12.003. 
            Epub 2016 Dec 8.
  PUBMED    27939849
REFERENCE   2  (bases 1 to 6022)
  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: "Metab 
            Eng."; date: "2017-01"; volume: "39:209-219. doi"; pages: " 
            10.1016/j.ymben.2016.12.003. Epub 2016 Dec 8"
COMMENT     Sequence Label: pILGFPU0 (R)
FEATURES             Location/Qualifiers
     source          1..6022
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             23..733
                     /codon_start=1
                     /product="yeast-enhanced green fluorescent protein"
                     /label=yeGFP
                     /note="codons optimized for translation in S. cerevisiae 
                     and C. albicans"
                     /translation="SKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLKF
                     ICTTGKLPVPWPTLVTTFGYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDGN
                     YKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKVN
                     FKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLEF
                     VTAAGITHGMDELYK"
     primer_bind     complement(1626..1642)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1626..1642)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1639..1661)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1650..1666
                     /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(1674..1680,1681..1698,1699..1704))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1719..1740
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1857..1874)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2028..2616)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2108..2127)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2787..3578,3579..3647))
                     /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     3410..3429
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3648..3752)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3820..3838
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(3876..3898)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     3998..4017
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4211..4233
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     4225..4242
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     4226..4242
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(5356..5389)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        5600..6013
                     /gene="S. cerevisiae TEF1"
                     /label=TEF1 promoter
                     /note="promoter for EF-1-alpha"
ORIGIN
        1 caaactcgag aaaatgggat cctctaaagg tgaagaatta ttcactggtg ttgtcccaat
       61 tttggttgaa ttagatggtg atgttaatgg tcacaaattt tctgtctccg gtgaaggtga
      121 aggtgatgct acttacggta aattgacctt aaaatttatt tgtactactg gtaaattgcc
      181 agttccatgg ccaaccttag tcactacttt cggttatggt gttcaatgtt ttgcgagata
      241 cccagatcat atgaaacaac atgacttttt caagtctgcc atgccagaag gttatgttca
      301 agaaagaact atttttttca aagatgacgg taactacaag accagagctg aagtcaagtt
      361 tgaaggtgat accttagtta atagaatcga attaaaaggt attgatttta aagaagatgg
      421 taacatttta ggtcacaaat tggaatacaa ctataactct cacaatgttt acatcatggc
      481 tgacaaacaa aagaatggta tcaaagttaa cttcaaaatt agacacaaca ttgaagatgg
      541 ttctgttcaa ttagctgacc attatcaaca aaatactcca attggtgatg gtccagtctt
      601 gttaccagac aaccattact tatccactca atctgcctta tccaaagatc caaacgaaaa
      661 gagagaccac atggtcttgt tagaatttgt tactgctgct ggtattaccc atggtatgga
      721 tgaattgtac aaaagatccg ttccaaccca acaagaagaa gagtacaagt tcaatatggt
      781 tttatctatc atcttgtccg ttcttcttgg gttttattat atatacactt tacacagagc
      841 ggctgtaggt tctgcatcca acttgaacat ttcgagaaag cttaccatat caaccccatc
      901 atgctctttc gaaaattcaa atagcacatc cattccttcg cccgcttcct catctcaaag
      961 ccacactcca atgagaaaca tgagctcact ctctgataac agcgttttca gccggaatat
     1021 ggaacaatca tcaccaatca ctccaagtat gtaccaattt ggtcagcagc agtcaaacag
     1081 tatatgtggt agcaccgtta gtgtgaatag tctggtgaat acaaataaca aacaaaggat
     1141 ctacgaacaa atcacgggtc ctaacagcaa taacgcaacc aatgattata ttgatttgct
     1201 aaacctaaat gagtctaaca aggaaaacca aaatcccgca acggcgcatt acctcaatgg
     1261 gggcccaccc aagacaagct tcattaacca tggaatgttc ccctcgccaa ctgggaccat
     1321 aaatagcggt aaatctagca gtgcctcatc tttaatttct tttggtatgg gcaataccca
     1381 agtaatatag actagtagga ctatttgcaa agggaaggga tgctaaggta gagggtgaac
     1441 gttacagaaa agcaggctgg gaagcatatt tgagaagatg cggccagcaa aactaaaaaa
     1501 ctgtattata agtaaatgca tgtatactaa actcacaaat tagagcttca atttaattat
     1561 atcagttatt acccgggaat ctcggtcgta atgatttaaa tgcatgcaag cttggcgtaa
     1621 tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc acacaacata
     1681 cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta actcacatta
     1741 attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa
     1801 tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg
     1861 ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag
     1921 gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa
     1981 ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc
     2041 cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca
     2101 ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg
     2161 accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct
     2221 catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt
     2281 gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag
     2341 tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc
     2401 agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac
     2461 actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga
     2521 gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc
     2581 aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg
     2641 gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca
     2701 aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt
     2761 atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca
     2821 gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg
     2881 atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca
     2941 ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt
     3001 cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt
     3061 agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca
     3121 cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca
     3181 tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga
     3241 agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact
     3301 gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga
     3361 gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg
     3421 ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc
     3481 tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga
     3541 tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat
     3601 gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt
     3661 caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt
     3721 atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac
     3781 gtctaagaaa ccattattat catgacatta acctataaaa ataggcgtat cacgaggccc
     3841 tttcgtctcg cgcgtttcgg tgatgacggt gaaaacctct gacacatgca gctcccggag
     3901 acggtcacag cttgtctgta agcggatgcc gggagcagac aagcccgtca gggcgcgtca
     3961 gcgggtgttg gcgggtgtcg gggctggctt aactatgcgg catcagagca gattgtactg
     4021 agagtgcacc atatgcggtg tgaaataccg cacagatgcg taaggagaaa ataccgcatc
     4081 aggcgccatt cgccattcag gctgcgcaac tgttgggaag ggcgatcggt gcgggcctct
     4141 tcgctattac gccagctggc gaaaggggga tgtgctgcaa ggcgattaag ttgggtaacg
     4201 ccagggtttt cccagtcacg acgttgtaaa acgacggcca gtgaattcat ttaaattttt
     4261 ttgattcggt aatctccgaa cagaaggaag aacgaaggaa ggagcacaga cttagattgg
     4321 tatatatacg catatgtagt gttgaagaaa catgaaattg cccagtattc ttaacccaac
     4381 tgcacagaac aaaaacctgc aggaaacgaa gataaaacat gtccacaaaa tcatatacca
     4441 gtagagctga gactcatgca agtccggttg catcgaaact tttacgttta atggatgaaa
     4501 agaagaccaa tttgtgtgct tctcttgacg ttcgttcgac tgatgagcta ttgaaacttg
     4561 ttgaaacgtt gggtccatac atttgccttt tgaaaacaca cgttgatatc ttggatgatt
     4621 tcagttatga gggtactgtc gttccattga aagcattggc agagaaatac aagttcttga
     4681 tatttgagga cagaaaattc gccgatatcg gtaacacagt caaattacaa tatacatcgg
     4741 gcgtttaccg tatcgcagaa tggtctgata tcaccaacgc ccacggggtt actggtgctg
     4801 gtattgttgc tggcttgaaa caaggtgcgc aagaggtcac caaagaacca aggggattat
     4861 tgatgcttgc tgaattgtct tccaagggtt ctctagcaca cggtgaatat actaagggta
     4921 ccgttgatat tgcaaagagt gataaagatt tcgttattgg gttcattgct cagaacgata
     4981 tgggaggaag agaagaaggg tttgattggc taatcatgac cccaggtgta ggtttagacg
     5041 acaaaggcga tgcattgggt cagcagtaca gaaccgtcga cgaagttgta agtggtggat
     5101 cagatatcat cattgttggc agaggacttt tcgccaaggg tagagatcct aaggttgaag
     5161 gtgaaagata cagaaatgct ggatgggaag cgtaccaaaa gagaatcagc gctccccatt
     5221 aattatacag gaaacttaat agaacaaatc acatatttaa tctaatagcc acctgcattg
     5281 gcacggtgca acactacttc aacttcatct tacaaaaaga tcacgtgatc tgttgtattg
     5341 ggatctctag acctaataac ttcgtatagc atacattata cgaagttata ttaagggttg
     5401 ctcgaccaga aagcgaccac ccaactttgg ctgataatag cgtataaaca atgcatactt
     5461 tgtacgttca aaatacaatg cagtagatat atttatgcat attacatata atacatatca
     5521 cataggaagc aacaggcgcg ttggactttt aattttcgag gaccgcgaat ccttacatca
     5581 cacccaatcc cccacaagtg atcccccaca caccatagct tcaaaatgtt tctactcctt
     5641 ttttactctt ccagattttc tcggactccg cgcatcgccg taccacttca aaacacccaa
     5701 gcacagcata ctaaatttcc cctctttctt cctctagggt gtcgttaatt acccgtacta
     5761 aaggtttgga aaagaaaaaa gagaccgcct cgtttctttt tcttcgtcga aaaaggcaat
     5821 aaaaattttt atcacgtttc tttttcttga aaattttttt ttttgatttt tttctctttc
     5881 gatgacctcc cattgatatt taagttaata aacggtcttc aatttctcaa gtttcagttt
     5941 catttttctt gttctattac aacttttttt acttcttgct cattagaaag aaagcatagc
     6001 aatctaatct aagttttaat ta
//
