LOCUS       sequence-436183-        8362 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Galactose inducible expression of UBQLN2.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8362)
  AUTHORS   Chavez Lab
  TITLE     Multiplex screening identifies broad modifiers of ALS-associated 
            proteotoxicity
  JOURNAL   unpublished
REFERENCE   2  (bases 1 to 8362)
  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            "unpublished"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: pAG426-GAL-UBQLN2
FEATURES             Location/Qualifiers
     source          1..8362
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     222..240
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(278..300)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     400..419
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        443..658
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     CDS             659..1462
                     /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(1593..2048)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1730..1751)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     1942..1961
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2174..2196
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     2188..2205
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     2189..2205
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2215..2234
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2215..2233
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     terminator      complement(2252..2499)
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     2465..2483
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     primer_bind     2501..2517
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     2502..2518
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    2551..2575
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             complement(4449..4472)
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     protein_bind    complement(4481..4505)
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(4513..4529)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4513..4529)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     promoter        complement(4544..4985)
                     /gene="S. cerevisiae GAL1"
                     /label=GAL1 promoter
                     /note="inducible promoter, regulated by Gal4"
     primer_bind     complement(4550..4573)
                     /label=GAL1
                     /note="S. cerevisiae GAL1 promoter, forward primer"
     protein_bind    4868..4985
                     /label=UAS
                     /bound_moiety="Gal4"
                     /note="upstream activating sequence mediating 
                     Gal4-dependent induction"
     primer_bind     complement(5009..5029)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(5009..5027)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(5048..5064)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5048..5064)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5061..5083)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5072..5088
                     /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(5096..5102,5103..5120,5121..5126))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5141..5162
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5279..5296)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5450..6038)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5530..5549)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6209..7000,7001..7069))
                     /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     6832..6851
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7070..7174)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      join(7201..8362,1..181)
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     protein_bind    complement(8164..8211)
                     /label=FRT
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="FLP-mediated recombination occurs in the 8-bp core 
                     sequence TCTAGAAA (Turan and Bode, 2011)."
ORIGIN
        1 cttatatgcg tctatttatg taggatgaaa ggtagtctag tacctcctgt gatattatcc
       61 cattccatgc ggggtatcgt atgcttcctt cagcactacc ctttagctgt tctatatgct
      121 gccactcctc aattggatta gtctcatcct tcaatgctat catttccttt gatattggat
      181 catactaaga aaccattatt atcatgacat taacctataa aaataggcgt atcacgaggc
      241 cctttcgtct cgcgcgtttc ggtgatgacg gtgaaaacct ctgacacatg cagctcccgg
      301 agacggtcac agcttgtctg taagcggatg ccgggagcag acaagcccgt cagggcgcgt
      361 cagcgggtgt tggcgggtgt cggggctggc ttaactatgc ggcatcagag cagattgtac
      421 tgagagtgca ccataccacc ttttcaattc atcatttttt ttttattctt ttttttgatt
      481 tcggtttcct tgaaattttt ttgattcggt aatctccgaa cagaaggaag aacgaaggaa
      541 ggagcacaga cttagattgg tatatatacg catatgtagt gttgaagaaa catgaaattg
      601 cccagtattc ttaacccaac tgcacagaac aaaaacctgc aggaaacgaa gataaatcat
      661 gtcgaaagct acatataagg aacgtgctgc tactcatcct agtcctgttg ctgccaagct
      721 atttaatatc atgcacgaaa agcaaacaaa cttgtgtgct tcattggatg ttcgtaccac
      781 caaggaatta ctggagttag ttgaagcatt aggtcccaaa atttgtttac taaaaacaca
      841 tgtggatatc ttgactgatt tttccatgga gggcacagtt aagccgctaa aggcattatc
      901 cgccaagtac aattttttac tcttcgaaga cagaaaattt gctgacattg gtaatacagt
      961 caaattgcag tactctgcgg gtgtatacag aatagcagaa tgggcagaca ttacgaatgc
     1021 acacggtgtg gtgggcccag gtattgttag cggtttgaag caggcggcag aagaagtaac
     1081 aaaggaacct agaggccttt tgatgttagc agaattgtca tgcaagggct ccctatctac
     1141 tggagaatat actaagggta ctgttgacat tgcgaagagc gacaaagatt ttgttatcgg
     1201 ctttattgct caaagagaca tgggtggaag agatgaaggt tacgattggt tgattatgac
     1261 acccggtgtg ggtttagatg acaagggaga cgcattgggt caacagtata gaaccgtgga
     1321 tgatgtggtc tctacaggat ctgacattat tattgttgga agaggactat ttgcaaaggg
     1381 aagggatgct aaggtagagg gtgaacgtta cagaaaagca ggctgggaag catatttgag
     1441 aagatgcggc cagcaaaact aaaaaactgt attataagta aatgcatgta tactaaactc
     1501 acaaattaga gcttcaattt aattatatca gttattaccc tatgcggtgt gaaataccgc
     1561 acagatgcgt aaggagaaaa taccgcatca ggaaattgta agcgttaata ttttgttaaa
     1621 attcgcgtta aatttttgtt aaatcagctc attttttaac caataggccg aaatcggcaa
     1681 aatcccttat aaatcaaaag aatagaccga gatagggttg agtgttgttc cagtttggaa
     1741 caagagtcca ctattaaaga acgtggactc caacgtcaaa gggcgaaaaa ccgtctatca
     1801 gggcgatggc ccactacgtg aaccatcacc ctaatcaagt tttttggggt cgaggtgccg
     1861 taaagcacta aatcggaacc ctaaagggag cccccgattt agagcttgac ggggaaagcc
     1921 ggcgaacgtg gcgagaaagg aagggaagaa agcgaaagga gcgggcgcta gggcgctggc
     1981 aagtgtagcg gtcacgctgc gcgtaaccac cacacccgcc gcgcttaatg cgccgctaca
     2041 gggcgcgtcc attcgccatt caggctgcgc aactgttggg aagggcgatc ggtgcgggcc
     2101 tcttcgctat tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta
     2161 acgccagggt tttcccagtc acgacgttgt aaaacgacgg ccagtgagcg cgcgtaatac
     2221 gactcactat agggcgaatt gggtaccggc cgcaaattaa agccttcgag cgtcccaaaa
     2281 ccttctcaag caaggttttc agtataatgt tacatgcgta cacgcgtctg tacagaaaaa
     2341 aaagaaaaat ttgaaatata aataacgttc ttaatactaa cataactata aaaaaataaa
     2401 tagggaccta gacttcaggt tgtctaactc cttccttttc ggttagagcg gatgtggggg
     2461 gagggcgtga atgtaagcgt gacataacta attacatgac tcgaggtcga cggtatcgat
     2521 aagcttgata tcgaattcct gcagcccatc accactttgt acaagaaagc tgggttttac
     2581 gatggctggg agcccagcag cctttcaatg gctgcattga tgtcgcctcc tgttgctatt
     2641 agggcctgca agtttgcttc acggtttaag aaccccattg cgttgagctg ttccagttgt
     2701 tgctgaaatc tgacttctgg attcggcagc tgtggagcat ttgctccagc cagggcctgc
     2761 accatttgct gaatgaactg ctggttgggt ccagattctg atgtaggact cgtggtttca
     2821 ctaggtgcag cgctggacac agtaggcccc gtgggaccac cagagccggt ggagccaggg
     2881 ggggctgcag ggccagtggg tcctatgggc ccaatggggc ctatgggggt aaaagggact
     2941 atagggccta tggggcctat gggggtgact gggcctacag ggcctatagc ggttcccagc
     3001 acccccaccc ccacacctgg agtgaagctc ggaatcaggc caggtgcttc agtggctaat
     3061 gtctgtagcc cctgctggat ctgcattaaa gcctgcattg ctcttgggtt tgacatggct
     3121 gatagtgtgt ctggattctg catctgctgc aggaaggctg ggagctgtgg ccgcatctgc
     3181 tcctgcagct gaggatttgc agtaaacagc gggctattca gcatcatctg tgcagccaaa
     3241 tctggattct ggctcagcga ctgcatcatg cttctcatgt agggcgccga cagcatattc
     3301 tgaatcagct gggggttttc agttatctgt tgcagcaggc tctgcatgcc tggggtacta
     3361 aagatgctgg cgacataatt agcggcagca acggtgttcc cagtagcatt gctggaacta
     3421 ttgccagacc cactaccagt gcttgtggtc gtgctggtag ttgcagaact ctgggtagct
     3481 ggcggtggtg cccatggatt gggtagtgga tcgcgatttt ctgtgcggga aggctgcgta
     3541 ccttccccag aggaggaact actccccacg gaggcaaatg gattaccccc aaactgctct
     3601 tgtgcggcat tcagcatcgg ctcttgaatg tcagtgtaca tgcgccgtaa agcattatag
     3661 ccacctggga tgctttctag attgctaaga gccaggtctt gatttctcat catctcttgc
     3721 atcatggctg gattcctggc aatttcgagt gtctgcctca ttatgtctgg gttgttgagc
     3781 aggtgactga tttctgggtt tctctgaatc aattgctgca tctgtggatt agccataatg
     3841 agctgcctca tcagatcggg attcgaaagc atgctctgaa caaagggatt ttccattatt
     3901 tggatcatca tctcagggct ggccataagc tgctgctgca tctggctctg gagctcagag
     3961 aagttggtcg agctcaagcc caggctgcta aggcctgcaa gtcctcccag gctccccaac
     4021 ccaaacgggt tgctatttgt ggaaataggt gtggagttac tcctgggagt cgacgccgag
     4081 gtagtgttag ttcccgcggc attgctaggc tgcgtggact ggccctgagg tcggttctgg
     4141 cttttgatga caaggtgaac agtcagccca tcatggatgc catgctggat caaggtatct
     4201 tgatctttta agatttttcc ggcaaaaatc agcactagct gatcggtttg ggatttgaag
     4261 cgtttcgaaa tcgcttcctt aaactgctga accgagctgt tctcgggcac cgcgaactcc
     4321 tctttctctt tgggagtctt caccgtgact ttgatgattt taggctcagc cggggcagca
     4381 gccgagcctt gggccgcagc agggccgcgg gaggggcgcg gggggccgct gctctcgcca
     4441 ttctcagcct tgtcatcgtc gtccttgtag tccattttgt agcctgcttt tttgtacaaa
     4501 cttgtgatgg gggatccact agttctagaa tccggggttt tttctccttg acgttaaagt
     4561 atagaggtat attaacaatt ttttgttgat acttttatta catttgaata agaagtaata
     4621 caaaccgaaa atgttgaaag tattagttaa agtggttatg cagtttttgc atttatatat
     4681 ctgttaatag atcaaaaatc atcgcttcgc tgattaatta ccccagaaat aaggctaaaa
     4741 aactaatcgc attatcatcc tatggttgtt aatttgattc gttcatttga aggtttgtgg
     4801 ggccaggtta ctgccaattt ttcctcttca taaccataaa agctagtatt gtagaatctt
     4861 tattgttcgg agcagtgcgg cgcgaggcac atctgcgttt caggaacgcg accggtgaag
     4921 acgaggacgc acggaggaga gtcttccttc ggagggctgt cacccgctcg gcggcttcta
     4981 atccgtacta gagctccagc ttttgttccc tttagtgagg gttaattgcg cgcttggcgt
     5041 aatcatggtc atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca
     5101 tacgagccgg aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat
     5161 taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt
     5221 aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct
     5281 cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa
     5341 aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa
     5401 aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc
     5461 tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga
     5521 caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc
     5581 cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt
     5641 ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct
     5701 gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg
     5761 agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta
     5821 gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct
     5881 acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa
     5941 gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt
     6001 gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta
     6061 cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat
     6121 caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa
     6181 gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct
     6241 cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta
     6301 cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct
     6361 caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg
     6421 gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa
     6481 gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt
     6541 cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta
     6601 catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca
     6661 gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta
     6721 ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct
     6781 gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg
     6841 cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac
     6901 tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact
     6961 gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa
     7021 atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt
     7081 ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat
     7141 gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg
     7201 aacgaagcat ctgtgcttca ttttgtagaa caaaaatgca acgcgagagc gctaattttt
     7261 caaacaaaga atctgagctg catttttaca gaacagaaat gcaacgcgaa agcgctattt
     7321 taccaacgaa gaatctgtgc ttcatttttg taaaacaaaa atgcaacgcg agagcgctaa
     7381 tttttcaaac aaagaatctg agctgcattt ttacagaaca gaaatgcaac gcgagagcgc
     7441 tattttacca acaaagaatc tatacttctt ttttgttcta caaaaatgca tcccgagagc
     7501 gctatttttc taacaaagca tcttagatta ctttttttct cctttgtgcg ctctataatg
     7561 cagtctcttg ataacttttt gcactgtagg tccgttaagg ttagaagaag gctactttgg
     7621 tgtctatttt ctcttccata aaaaaagcct gactccactt cccgcgttta ctgattacta
     7681 gcgaagctgc gggtgcattt tttcaagata aaggcatccc cgattatatt ctataccgat
     7741 gtggattgcg catactttgt gaacagaaag tgatagcgtt gatgattctt cattggtcag
     7801 aaaattatga acggtttctt ctattttgtc tctatatact acgtatagga aatgtttaca
     7861 ttttcgtatt gttttcgatt cactctatga atagttctta ctacaatttt tttgtctaaa
     7921 gagtaatact agagataaac ataaaaaatg tagaggtcga gtttagatgc aagttcaagg
     7981 agcgaaaggt ggatgggtag gttatatagg gatatagcac agagatatat agcaaagaga
     8041 tacttttgag caatgtttgt ggaagcggta ttcgcaatat tttagtagct cgttacagtc
     8101 cggtgcgttt ttggtttttt gaaagtgcgt cttcagagcg cttttggttt tcaaaagcgc
     8161 tctgaagttc ctatactttc tagagaatag gaacttcgga ataggaactt caaagcgttt
     8221 ccgaaaacga gcgcttccga aaatgcaacg cgagctgcgc acatacagct cactgttcac
     8281 gtcgcaccta tatctgcgtg ttgcctgtat atatatatac atgagaagaa cggcatagtg
     8341 cgtgtttatg cttaaatgcg ta
//
