LOCUS       sequence-330313-        6900 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Integrative plasmid for cassette TDH3p:his6-mKate2-MBP-IDP1t.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6900)
  AUTHORS   Romero-Suarez D, Wulff T, Rong Y, Jakociu Nas T, Yuzawa S, Keasling 
            JD, Jensen MK
  TITLE     A Reporter System for Cytosolic Protein Aggregates in Yeast.
  JOURNAL   ACS Synth Biol. 2021 Feb 12. doi: 10.1021/acssynbio.0c00476.
  PUBMED    33577304
REFERENCE   2  (bases 1 to 6900)
  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: "ACS Synth 
            Biol. 2021 Feb 12. doi: 10.1021/acssynbio.0c00476."
COMMENT     Sequence Label: pYR11 (XII-5:TDH3p:his6-mKate2-MBP-IDP1t)
FEATURES             Location/Qualifiers
     source          1..6900
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        32..698
                     /gene="S. cerevisiae TDH3"
                     /label=GAP promoter
                     /note="promoter for glyceraldehyde-3-phosphate 
                     dehydrogenase; also known as the TDH3 promoter"
     primer_bind     568..591
                     /label=GPDpro-F
                     /note="S. cerevisiae GPD promoter, forward primer"
     CDS             717..734
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     protein_bind    complement(3629..3662)
                     /label=FRT (minimal)
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="supports FLP-mediated excision but not integration 
                     (Turan and Bode, 2011)"
     primer_bind     complement(4132..4149)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4132..4148)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4247..4264)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4418..5006)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(join(5128..5919,5920..5988))
                     /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     5751..5770
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5989..6080)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6122..6141
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     6147..6166
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     protein_bind    6373..6406
                     /label=FRT (minimal)
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="supports FLP-mediated excision but not integration 
                     (Turan and Bode, 2011)"
ORIGIN
        1 ataaaaaaca cgctttttca gttcgagttt atcattatca atactgccat ttcaaagaat
       61 acgtaaataa ttaatagtag tgattttcct aactttattt agtcaaaaaa ttagcctttt
      121 aattctgctg taacccgtac atgcccaaaa tagggggcgg gttacacaga atatataaca
      181 tcgtaggtgt ctgggtgaac agtttattcc tggcatccac taaatataat ggagcccgct
      241 ttttaagctg gcatccagaa aaaaaaagaa tcccagcacc aaaatattgt tttcttcacc
      301 aaccatcagt tcataggtcc attctcttag cgcaactaca gagaacaggg gcacaaacag
      361 gcaaaaaacg ggcacaacct caatggagtg atgcaacctg cctggagtaa atgatgacac
      421 aaggcaattg acccacgcat gtatctatct cattttctta caccttctat taccttctgc
      481 tctctctgat ttggaaaaag ctgaaaaaaa aggttgaaac cagttccctg aaattattcc
      541 cctacttgac taataagtat ataaagacgg taggtattga ttgtaattct gtaaatctat
      601 ttcttaaact tcttaaattc tacttttata gttagtcttt tttttagttt taaaacacca
      661 agaacttagt ttcgaataaa cacacataaa caaacaaaaa aacaatgggt agcagccacc
      721 accaccacca ccacggcggt ggcggttcta tggtttctga actcatcaag gaaaacatgc
      781 acatgaaact ttacatggaa ggtactgtga acaatcatca ttttaagtgt acatccgagg
      841 gtgaaggcaa accttacgaa ggaactcaaa ctatgagaat taaagctgta gaaggtggac
      901 cattaccttt tgcatttgat atcttggcaa catcattcat gtatgggagc aagacattca
      961 taaaccatac tcaaggtata ccagactttt tcaaacagag ttttccagag ggttttacat
     1021 gggaaagagt aacaacgtac gaggatggag gtgtattgac agccactcaa gacacatcac
     1081 ttcaagatgg gtgtttaatc tacaatgtca agattagagg cgtcaatttc ccttctaatg
     1141 gtccagttat gcagaaaaag acattaggct gggaagcgtc aaccgaaacc ctgtaccctg
     1201 ctgatggtgg cctagaaggc agagctgaca tggcccttaa actggttggt ggagggcatc
     1261 taatctgcaa tttgaaaacc acttatcgtt ctaaaaagcc agccaaaaac ctaaagatgc
     1321 caggtgttta ctacgtcgac cgaagattag aaaggattaa agaggctgat aaagagactt
     1381 atgttgaaca acacgaagtg gcagtggcta gatactgtga tttgccatct aagttgggac
     1441 acagagacgg tggatccggg ggtggtagca cttctatgaa gatcaaaacg ggcgcaagaa
     1501 tcttggcgct aagtgcgttg actacaatga tgttctctgc gtcagccctt gccaaaattg
     1561 aggagggtaa gttggtcatc tggataaacg gagacaaggg ctataacggg ctggcggaag
     1621 tgggaaagaa attcgagaag gataccggta taaaagtaac agttgaacat cctgataaat
     1681 tggaggagaa atttcctcaa gtcgccgcaa cgggggacgg cccagatatc atcttctggg
     1741 cccatgatag atttggtggg tatgcacaaa gcggcctact agctgaaatc acaccagata
     1801 aggcattcca agataagtta tatccattca catgggacgc ggtacgttat aatggtaagc
     1861 taatcgcgta ccctatagca gtcgaagctc tttccctgat ttacaataag gatctgcttc
     1921 ccaacccacc caagacgtgg gaggaaatac cggcattaga taaggagcta aaagcgaagg
     1981 gaaaatccgc tttaatgttt aacctgcaag aaccatattt tacatggccc ctaatcgccg
     2041 ccgacggagg gtatgcattt aagtatgaaa acggaaaata tgatataaaa gatgtcggcg
     2101 tagataacgc gggagccaag gcggggttaa ctttcctagt cgatctaatc aagaacaagc
     2161 acatgaacgc cgatacggat tattctatcg cagaagctgc ttttaataaa ggagaaaccg
     2221 caatgactat caacgggccg tgggcctgga gtaacataga tacgtctaag gtgaactacg
     2281 gagtaaccgt tctgccgacg tttaaaggtc aaccttcaaa acccttcgtc ggcgtgttaa
     2341 gcgcagggat taacgcggcg agccctaata aggaacttgc caaagagttc ctggaaaatt
     2401 atcttcttac cgacgaagga ttggaggcag taaataaaga taagccactt ggagctgttg
     2461 cgctgaagag ctatgaggag gagttagcga aggatccccg tattgcagca acaatggaga
     2521 acgcgcaaaa gggagagatt atgccgaaca tcccacagat gtctgctttc tggtatgccg
     2581 taaggacggc cgtcattaat gcagcatccg gcaggcagac cgtcgatgag gctctaaagg
     2641 acgcacagac atgaagcttc gaatttacgt agcccaatct accacttttt tttttcattt
     2701 tttaaagtgt tatacttagt tatgctctag gataatgaac tacttttttt tttttttttt
     2761 tttactgtta tcataaatat atatacctta ttgttgtttg caaccgtcgg ttaattcctt
     2821 atcaaggttc cccaagttcg gatcattacc atcaatttcc aacattttca tgagttcttc
     2881 ttcttcatta ccgtgtttta gggggctgtt gcaactcaga agtttgacag caagcaagtt
     2941 catcattcga actagcctta ttgttttagt tcagtgacag cgaactgccg tactcgatgc
     3001 tttatttctc acggtagagc ggaagaacag ataggggcag cgtgagaaga gttagaaagt
     3061 aaatttttat cacgtctgaa gtattcttat tcataggaaa ttttgcaagg ttttttagct
     3121 caataacggg ctaagttata taaggtgttc acgcgatttt cttgttatgt atacctcttc
     3181 tctgaggaat ggtactactg tcctgatgta ggctccttaa attggtgggc aagaataact
     3241 tatcgatatt ttgtatattg gtcttggagt tcaccacgta atgcctgttt aagaccatca
     3301 gttaactcta gtattatttg gtcttggcta ctggccgttt gctattattc aagtcttttg
     3361 tgccttcccg tcgggtaagg gagttattta gggatacaga atctaacgaa aactaaatct
     3421 caatgattaa ctctatttaa tccttttttg aaaggcaaaa gaggtccctt gttcacttac
     3481 aacgttctta gccaaattcg cttatcactt actacttcac gatatacaga agtaaaaaca
     3541 tataaaaaga tgtctgtttg tttagccatc acaaaaggta tcgcagatta agacctcagc
     3601 gcggccgcaa atttaaataa aatgaagtga agttcctata ctttctagag aataggaact
     3661 tctatagtga gtcgaataag ggcgacacaa aatttattct aaatgcataa taaatactga
     3721 taacatctta tagtttgtat tatattttgt attatcgttg acatgtataa ttttgatatc
     3781 aaaaactgat tttcccttta ttattttcga gatttatttt cttaattctc tttaacaaac
     3841 tagaaatatt gtatatacaa aaaatcataa ataatagatg aatagtttaa ttataggtgt
     3901 tcatcaatcg aaaaagcaac gtatcttatt taaagtgcgt tgcttttttc tcatttataa
     3961 ggttaaataa ttctcatata tcaagcaaag tgacaggcgc ccttaaatat tctgacaaat
     4021 gctctttccc taaactcccc ccataaaaaa acccgccgaa gcgggttttt acgttatttg
     4081 cggattaacg attactcgtt atcagaaccg cccagggggc ccgagcttaa gactggccgt
     4141 cgttttacaa cacagaaaga gtttgtagaa acgcaaaaag gccatccgtc aggggccttc
     4201 tgcttagttt gatgcctggc agttccctac tctcgccttc cgcttcctcg ctcactgact
     4261 cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac
     4321 ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa
     4381 aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg
     4441 acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa
     4501 gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc
     4561 ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac
     4621 gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac
     4681 cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg
     4741 taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt
     4801 atgtaggcgg tgctacagag ttcttgaagt ggtgggctaa ctacggctac actagaagaa
     4861 cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct
     4921 cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga
     4981 ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg
     5041 ctcagtggaa cgacgcgcgc gtaactcacg ttaagggatt ttggtcatga gcttgcgccg
     5101 tcccgtcaag tcagcgtaat gctctgctta ccaatgctta atcagtgagg cacctatctc
     5161 agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac
     5221 gatacgggag ggcttaccat ctggccccag cgctgcgatg ataccgcgag aaccacgctc
     5281 accggctccg gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg
     5341 tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag
     5401 tagttcgcca gttaatagtt tgcgcaacgt tgttgccatc gctacaggca tcgtggtgtc
     5461 acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac
     5521 atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag
     5581 aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac
     5641 tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg
     5701 agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc
     5761 gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact
     5821 ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg
     5881 atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa
     5941 tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatat tcttcctttt
     6001 tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg
     6061 tatttagaaa aataaacaaa taggggtcag tgttacaacc aattaaccaa ttctgaacat
     6121 tatcgcgagc ccatttatac ctgaatatgg ctcataacac cccttgtttg cctggcggca
     6181 gtagcgcggt ggtcccacct gaccccatgc cgaactcaga agtgaaacgc cgtagcgccg
     6241 atggtagtgt ggggactccc catgcgagag tagggaactg ccaggcatca aataaaacga
     6301 aaggctcagt cgaaagactg ggcctttcgc ccgggctaat tatggggtgt cgcccttatt
     6361 cgactctata gtgaagttcc tattctctag aaagtatagg aacttctgaa gtggggattt
     6421 aaatgcggcc gcgctgaggg tttaatcaat ctggcggctt gagttctcaa catgttttat
     6481 tttttactta tattgctggt agggtaaaaa aatataactc ctaggaatag gttgtctata
     6541 tgtttttgtc ttgcttctat aattgtaaca aacaaggaaa gggaaaatac tgggtgtaaa
     6601 agccattgag tcaagttagg tcatcccttt tatacaaaat ttttcaattt tttttccaag
     6661 attcttgtac gattaattat tttttttttg cgtcctacag cgtgatgaaa atttcgcctg
     6721 ctgcaagatg agcgggaacg ggcgaaatgt gcacgcgcac aacttacgaa acgcggatga
     6781 gtcactgaca gccaccgcag aggttctgac tcctactgag ctctattgga ggtggcagaa
     6841 ccggtaccgg aggagaccgc tataaccggt ttgaatttat tgtcacagtg tcacatcagc
//
