LOCUS       sequence-450959-        7755 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Fluorescent reporter for cytoplasmic aggregates.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7755)
  TITLE     Budding yeast expression plasmid for induced production of 
            fluorescently tagged cytoplasmic protein aggregates
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 7755)
  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: 
            "Unpublished"
COMMENT     Sequence Label: CUP1-∆ssCPY*-mScarletI
FEATURES             Location/Qualifiers
     source          1..7755
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(735..753)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     promoter        complement(919..937)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(958..974)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(958..974)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(971..993)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    982..998
                     /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(1006..1012,1013..1030,1031..1036))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1051..1072
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1189..1206)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1360..1948)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1440..1459)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2119..2910,2911..2979))
                     /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     2742..2761
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2980..3084)
                     /gene="bla"
                     /label=AmpR promoter
     misc_feature    3121..3624
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     primer_bind     3665..3683
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(3721..3743)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     3843..3862
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        3886..4101
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     CDS             4102..4905
                     /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      complement(5036..5491)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5173..5194)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5385..5404
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5617..5639
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     5631..5648
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     5632..5648
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5655..5674
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        5655..5673
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        5837..6071
                     /gene="S. cerevisiae CUP1"
                     /label=CUP1 promoter
                     /note="inducible metallothionein promoter, regulated by 
                     Cu2+"
ORIGIN
        1 gaagggcggc cttacgaagg tacgcagact gctaaattga aagttaccaa aggtggccca
       61 cttccattta gttgggatat tttaagccca caattcatgt atgggtccag agctttcatc
      121 aaacaccctg ctgatatacc tgactattac aagcagagct tcccagaagg gttcaagtgg
      181 gaaagagtaa tgaactttga agacggagga gctgtcaccg tgacgcagga taccagtctt
      241 gaagacggca cgctgattta taaggtcaag ctcagaggga caaattttcc tcctgacggg
      301 cctgttatgc agaaaaaaac gatgggttgg gaggcatcaa cggagcgtct ctatccagag
      361 gacggtgttt taaagggaga tatcaaaatg gcactccgtc tcaaggatgg aggcagatac
      421 ctcgctgact ttaagaccac atacaaagcg aaaaaacctg ttcaaatgcc aggggcgtat
      481 aacgtggatc gtaagctgga cattaccagc cataacgaag attacacagt ggtggagcag
      541 tatgagagaa gtgaaggccg ccattccacg ggtggtatgg atgagctgta taaataaaag
      601 ctttgccata gaatatctgg atccggtacc gggttgaaga aggaaaaaga cagaaacgac
      661 ttaattacct acttgaaaaa agcctgtgag taaacaggcc ccttttcctt tgtcgatatc
      721 atgtaattag ttatgtcacg cttacattca cgccctcccc ccacatccgc tctaaccgaa
      781 aaggaaggag ttagacaacc tgaagtctag gtccctattt atttttttat agttatgtta
      841 gtattaagaa cgttatttat atttcaaatt tttctttttt ttctgtacaa acgcgtgtac
      901 ggtacccagc ttttgttccc tttagtgagg gttaattccg agcttggcgt aatcatggtc
      961 atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg
     1021 aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt
     1081 gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg
     1141 ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga
     1201 ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat
     1261 acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca
     1321 aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc
     1381 tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata
     1441 aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc
     1501 gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc
     1561 acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga
     1621 accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc
     1681 ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag
     1741 gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag
     1801 aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag
     1861 ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca
     1921 gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga
     1981 cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat
     2041 cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga
     2101 gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg
     2161 tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga
     2221 gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc
     2281 agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac
     2341 tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc
     2401 agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc
     2461 gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc
     2521 catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt
     2581 ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc
     2641 atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg
     2701 tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag
     2761 cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat
     2821 cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc
     2881 atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa
     2941 aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta
     3001 ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa
     3061 aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg ggtccttttc
     3121 atcacgtgct ataaaaataa ttataattta aattttttaa tataaatata taaattaaaa
     3181 atagaaagta aaaaaagaaa ttaaagaaaa aatagttttt gttttccgaa gatgtaaaag
     3241 actctagggg gatcgccaac aaatactacc ttttatcttg ctcttcctgc tctcaggtat
     3301 taatgccgaa ttgtttcatc ttgtctgtgt agaagaccac acacgaaaat cctgtgattt
     3361 tacattttac ttatcgttaa tcgaatgtat atctatttaa tctgcttttc ttgtctaata
     3421 aatatatatg taaagtacgc tttttgttga aattttttaa acctttgttt attttttttt
     3481 cttcattccg taactcttct accttcttta tttactttct aaaatccaaa tacaaaacat
     3541 aaaaataaat aaacacagag taaattccca aattattcca tcattaaaag atacgaggcg
     3601 cgtgtaagtt acaggcaagc gatccgtcta agaaaccatt attatcatga cattaaccta
     3661 taaaaatagg cgtatcacga ggccctttcg tctcgcgcgt ttcggtgatg acggtgaaaa
     3721 cctctgacac atgcagctcc cggagacggt cacagcttgt ctgtaagcgg atgccgggag
     3781 cagacaagcc cgtcagggcg cgtcagcggg tgttggcggg tgtcggggct ggcttaacta
     3841 tgcggcatca gagcagattg tactgagagt gcaccatacc accttttcaa ttcatcattt
     3901 tttttttatt cttttttttg atttcggttt ccttgaaatt tttttgattc ggtaatctcc
     3961 gaacagaagg aagaacgaag gaaggagcac agacttagat tggtatatat acgcatatgt
     4021 agtgttgaag aaacatgaaa ttgcccagta ttcttaaccc aactgcacag aacaaaaacc
     4081 tgcaggaaac gaagataaat catgtcgaaa gctacatata aggaacgtgc tgctactcat
     4141 cctagtcctg ttgctgccaa gctatttaat atcatgcacg aaaagcaaac aaacttgtgt
     4201 gcttcattgg atgttcgtac caccaaggaa ttactggagt tagttgaagc attaggtccc
     4261 aaaatttgtt tactaaaaac acatgtggat atcttgactg atttttccat ggagggcaca
     4321 gttaagccgc taaaggcatt atccgccaag tacaattttt tactcttcga agacagaaaa
     4381 tttgctgaca ttggtaatac agtcaaattg cagtactctg cgggtgtata cagaatagca
     4441 gaatgggcag acattacgaa tgcacacggt gtggtgggcc caggtattgt tagcggtttg
     4501 aagcaggcgg cagaagaagt aacaaaggaa cctagaggcc ttttgatgtt agcagaattg
     4561 tcatgcaagg gctccctatc tactggagaa tatactaagg gtactgttga cattgcgaag
     4621 agcgacaaag attttgttat cggctttatt gctcaaagag acatgggtgg aagagatgaa
     4681 ggttacgatt ggttgattat gacacccggt gtgggtttag atgacaaggg agacgcattg
     4741 ggtcaacagt atagaaccgt ggatgatgtg gtctctacag gatctgacat tattattgtt
     4801 ggaagaggac tatttgcaaa gggaagggat gctaaggtag agggtgaacg ttacagaaaa
     4861 gcaggctggg aagcatattt gagaagatgc ggccagcaaa actaaaaaac tgtattataa
     4921 gtaaatgcat gtatactaaa ctcacaaatt agagcttcaa tttaattata tcagttatta
     4981 ccctatgcgg tgtgaaatac cgcacagatg cgtaaggaga aaataccgca tcaggaaatt
     5041 gtaagcgtta atattttgtt aaaattcgcg ttaaattttt gttaaatcag ctcatttttt
     5101 aaccaatagg ccgaaatcgg caaaatccct tataaatcaa aagaatagac cgagataggg
     5161 ttgagtgttg ttccagtttg gaacaagagt ccactattaa agaacgtgga ctccaacgtc
     5221 aaagggcgaa aaaccgtcta tcagggcgat ggcccactac gtgaaccatc accctaatca
     5281 agttttttgg ggtcgaggtg ccgtaaagca ctaaatcgga accctaaagg gagcccccga
     5341 tttagagctt gacggggaaa gccggcgaac gtggcgagaa aggaagggaa gaaagcgaaa
     5401 ggagcgggcg ctagggcgct ggcaagtgta gcggtcacgc tgcgcgtaac caccacaccc
     5461 gccgcgctta atgcgccgct acagggcgcg tccattcgcc attcaggctg cgcaactgtt
     5521 gggaagggcg atcggtgcgg gcctcttcgc tattacgcca gctggcgaaa gggggatgtg
     5581 ctgcaaggcg attaagttgg gtaacgccag ggttttccca gtcacgacgt tgtaaaacga
     5641 cggccagtga attgtaatac gactcactat agggcgaatt ggagctccac cgcggtggcg
     5701 gccgcatgtt catgtatgta tctgtattta aaacactttt gtattatttt tcctcatata
     5761 tgtgtatagg tttatacgga tgatttaatt attacttcac caccctttat ttcaggctga
     5821 tatcttagcc ttgttactag ttagaaaaag acatttttgc tgtcagtcac tgtcaagaga
     5881 ttcttttgct ggcatttctt ctagaagcaa aaagagcgat gcgtcttttc cgctgaaccg
     5941 ttccagcaaa aaagactacc aacgcaatat ggattgtcag aatcatataa aagagaagca
     6001 aataactcct tgtcttgtgt caattgcatt ataatatctt cttgttagtg caatatcata
     6061 tagaagtcat cgaaatagat attaagaaaa acaaactgta acgaattcat gatctcattg
     6121 caaagaccgt tgggtctaga taaggacgtt ttgctgcaag ctgcggaaaa atttggtttg
     6181 gacctcgacc tggatcatct cttgaaggag ttggactcca atgtattgga cgcttgggcc
     6241 caaatagagc atttgtaccc aaaccaggtt atgagccttg aaacttccac taagccaaaa
     6301 ttccctgaag caatcaaaac gaagaaagac tgggactttg tggtcaagaa tgacgcaatt
     6361 gaaaactatc agcttcgtgt caacaagatt aaggacccta aaatcctggg cattgaccca
     6421 aatgtcacac agtacacggg ttacttggat gtggaagacg aggacaagca tttcttcttt
     6481 tggacttttg aaagtagaaa cgatcctgca aaggatccgg tcatcctttg gttgaacggg
     6541 ggtccaggtt gttcttcact aaccgggctg ttctttgaat taggaccctc atccattgga
     6601 cctgatttga aacccatcgg gaacccttac tcttggaaca gcaatgccac cgtgatcttc
     6661 cttgaccagc ctgtcaacgt tgggttctcg tattccgggt cctcaggtgt ttccaacact
     6721 gtcgccgctg gtaaggatgt ctataacttc ttggagttgt tcttcgatca gttccctgaa
     6781 tacgtcaaca agggccaaga tttccacatc gctagggaat cctacgccgg ccattacatc
     6841 cctgtttttg cctctgaaat tttgtctcac aaggacagaa acttcaactt aacctccgtc
     6901 ttgatcggaa atggcctcac tgacccattg actcagtata actattacga accaatggcc
     6961 tgtggtgaag gtggcgaacc atctgttttg ccctcggagg aatgctctgc tatggaagac
     7021 tctttggaac gttgtttggg cttgatcgag tcgtgctatg actcgcaatc cgtctggtcc
     7081 tgtgttccag ctaccattta ttgtaataac gcccaattgg ctccttacca acgtaccggc
     7141 agaaacgttt acgatatcag gaaggattgt gaaggtggca atttgtgcta cccaacgtta
     7201 caagatatcg acgactactt aaaccaggac tacgtcaaag aagctgtcgg tgcggaggtt
     7261 gaccactacg aatcctgtaa cttcgatatc aacagaaatt tcctgtttgc gggtgattgg
     7321 atgaagcctt accacaccgc cgtaacagat cttttgaatc aagacctacc cattctggta
     7381 tatgcaggcg ataaagattt catctgtaac tggttgggta ataaggcgtg gacggatgtc
     7441 ttgccatgga agtacgacga agaatttgca agccaaaaag tacgtaactg gactgcttct
     7501 atcaccgacg aggtcgctgg tgaagtcaaa tcctacaagc acttcaccta tttgagagtc
     7561 ttcaatggtg gccacatggt tccatttgac gtccctgaaa acgccttaag tatggttaac
     7621 gaatggatcc acggtggttt ctccttactc gagatggtct ccaaaggaga ggccgtaatc
     7681 aaagagttca tgagattcaa agttcacatg gaggggtcta tgaacggcca tgagttcgag
     7741 atagaaggag agggc
//
