LOCUS       sequence-430960-        6879 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Galactose inducible expression of TDP-43-Q331K.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6879)
  AUTHORS   Chavez Lab
  TITLE     Multiplex screening identifies broad modifiers of ALS-associated 
            proteotoxicity
  JOURNAL   unpublished
REFERENCE   2  (bases 1 to 6879)
  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            "unpublished"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: pAG416-GAL-TDP-43-Q331K
FEATURES             Location/Qualifiers
     source          1..6879
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(16..457)
                     /gene="S. cerevisiae GAL1"
                     /label=GAL1 promoter
                     /note="inducible promoter, regulated by Gal4"
     primer_bind     complement(22..45)
                     /label=GAL1
                     /note="S. cerevisiae GAL1 promoter, forward primer"
     protein_bind    340..457
                     /label=UAS
                     /bound_moiety="Gal4"
                     /note="upstream activating sequence mediating 
                     Gal4-dependent induction"
     primer_bind     complement(481..501)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(481..499)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(520..536)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(520..536)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(533..555)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    544..560
                     /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(568..574,575..592,593..598))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    613..634
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(751..768)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(922..1510)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1002..1021)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1681..2472,2473..2541))
                     /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     2304..2323
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2542..2646)
                     /gene="bla"
                     /label=AmpR promoter
     misc_feature    2683..3186
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     primer_bind     3227..3245
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(3283..3305)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     3405..3424
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        3448..3663
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     CDS             3664..4467
                     /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(4598..5053)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4735..4756)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4947..4966
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5179..5201
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     5193..5210
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     5194..5210
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5220..5239
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        5220..5238
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     terminator      complement(5257..5504)
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     5470..5488
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     primer_bind     5506..5522
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     5507..5523
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    5556..5580
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(6832..6856)
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(join(6864..6879,1))
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(join(6864..6879,1))
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
ORIGIN
        1 aatccggggt tttttctcct tgacgttaaa gtatagaggt atattaacaa ttttttgttg
       61 atacttttat tacatttgaa taagaagtaa tacaaaccga aaatgttgaa agtattagtt
      121 aaagtggtta tgcagttttt gcatttatat atctgttaat agatcaaaaa tcatcgcttc
      181 gctgattaat taccccagaa ataaggctaa aaaactaatc gcattatcat cctatggttg
      241 ttaatttgat tcgttcattt gaaggtttgt ggggccaggt tactgccaat ttttcctctt
      301 cataaccata aaagctagta ttgtagaatc tttattgttc ggagcagtgc ggcgcgaggc
      361 acatctgcgt ttcaggaacg cgaccggtga agacgaggac gcacggagga gagtcttcct
      421 tcggagggct gtcacccgct cggcggcttc taatccgtac tagagctcca gcttttgttc
      481 cctttagtga gggttaattg cgcgcttggc gtaatcatgg tcatagctgt ttcctgtgtg
      541 aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc
      601 ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt
      661 ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg
      721 cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt
      781 tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc
      841 aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa
      901 aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa
      961 tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc
     1021 ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc
     1081 cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag
     1141 ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga
     1201 ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc
     1261 gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac
     1321 agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg
     1381 cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca
     1441 aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa
     1501 aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa
     1561 ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt
     1621 aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag
     1681 ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat
     1741 agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc
     1801 cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa
     1861 ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca
     1921 gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa
     1981 cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt
     2041 cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc
     2101 ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact
     2161 catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc
     2221 tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg
     2281 ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct
     2341 catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc
     2401 cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag
     2461 cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac
     2521 acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg
     2581 ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt
     2641 tccgcgcaca tttccccgaa aagtgccacc tgggtccttt tcatcacgtg ctataaaaat
     2701 aattataatt taaatttttt aatataaata tataaattaa aaatagaaag taaaaaaaga
     2761 aattaaagaa aaaatagttt ttgttttccg aagatgtaaa agactctagg gggatcgcca
     2821 acaaatacta ccttttatct tgctcttcct gctctcaggt attaatgccg aattgtttca
     2881 tcttgtctgt gtagaagacc acacacgaaa atcctgtgat tttacatttt acttatcgtt
     2941 aatcgaatgt atatctattt aatctgcttt tcttgtctaa taaatatata tgtaaagtac
     3001 gctttttgtt gaaatttttt aaacctttgt ttattttttt ttcttcattc cgtaactctt
     3061 ctaccttctt tatttacttt ctaaaatcca aatacaaaac ataaaaataa ataaacacag
     3121 agtaaattcc caaattattc catcattaaa agatacgagg cgcgtgtaag ttacaggcaa
     3181 gcgatccgtc taagaaacca ttattatcat gacattaacc tataaaaata ggcgtatcac
     3241 gaggcccttt cgtctcgcgc gtttcggtga tgacggtgaa aacctctgac acatgcagct
     3301 cccggagacg gtcacagctt gtctgtaagc ggatgccggg agcagacaag cccgtcaggg
     3361 cgcgtcagcg ggtgttggcg ggtgtcgggg ctggcttaac tatgcggcat cagagcagat
     3421 tgtactgaga gtgcaccata ccaccttttc aattcatcat ttttttttta ttcttttttt
     3481 tgatttcggt ttccttgaaa tttttttgat tcggtaatct ccgaacagaa ggaagaacga
     3541 aggaaggagc acagacttag attggtatat atacgcatat gtagtgttga agaaacatga
     3601 aattgcccag tattcttaac ccaactgcac agaacaaaaa cctgcaggaa acgaagataa
     3661 atcatgtcga aagctacata taaggaacgt gctgctactc atcctagtcc tgttgctgcc
     3721 aagctattta atatcatgca cgaaaagcaa acaaacttgt gtgcttcatt ggatgttcgt
     3781 accaccaagg aattactgga gttagttgaa gcattaggtc ccaaaatttg tttactaaaa
     3841 acacatgtgg atatcttgac tgatttttcc atggagggca cagttaagcc gctaaaggca
     3901 ttatccgcca agtacaattt tttactcttc gaagacagaa aatttgctga cattggtaat
     3961 acagtcaaat tgcagtactc tgcgggtgta tacagaatag cagaatgggc agacattacg
     4021 aatgcacacg gtgtggtggg cccaggtatt gttagcggtt tgaagcaggc ggcagaagaa
     4081 gtaacaaagg aacctagagg ccttttgatg ttagcagaat tgtcatgcaa gggctcccta
     4141 tctactggag aatatactaa gggtactgtt gacattgcga agagcgacaa agattttgtt
     4201 atcggcttta ttgctcaaag agacatgggt ggaagagatg aaggttacga ttggttgatt
     4261 atgacacccg gtgtgggttt agatgacaag ggagacgcat tgggtcaaca gtatagaacc
     4321 gtggatgatg tggtctctac aggatctgac attattattg ttggaagagg actatttgca
     4381 aagggaaggg atgctaaggt agagggtgaa cgttacagaa aagcaggctg ggaagcatat
     4441 ttgagaagat gcggccagca aaactaaaaa actgtattat aagtaaatgc atgtatacta
     4501 aactcacaaa ttagagcttc aatttaatta tatcagttat taccctatgc ggtgtgaaat
     4561 accgcacaga tgcgtaagga gaaaataccg catcaggaaa ttgtaagcgt taatattttg
     4621 ttaaaattcg cgttaaattt ttgttaaatc agctcatttt ttaaccaata ggccgaaatc
     4681 ggcaaaatcc cttataaatc aaaagaatag accgagatag ggttgagtgt tgttccagtt
     4741 tggaacaaga gtccactatt aaagaacgtg gactccaacg tcaaagggcg aaaaaccgtc
     4801 tatcagggcg atggcccact acgtgaacca tcaccctaat caagtttttt ggggtcgagg
     4861 tgccgtaaag cactaaatcg gaaccctaaa gggagccccc gatttagagc ttgacgggga
     4921 aagccggcga acgtggcgag aaaggaaggg aagaaagcga aaggagcggg cgctagggcg
     4981 ctggcaagtg tagcggtcac gctgcgcgta accaccacac ccgccgcgct taatgcgccg
     5041 ctacagggcg cgtccattcg ccattcaggc tgcgcaactg ttgggaaggg cgatcggtgc
     5101 gggcctcttc gctattacgc cagctggcga aagggggatg tgctgcaagg cgattaagtt
     5161 gggtaacgcc agggttttcc cagtcacgac gttgtaaaac gacggccagt gagcgcgcgt
     5221 aatacgactc actatagggc gaattgggta ccggccgcaa attaaagcct tcgagcgtcc
     5281 caaaaccttc tcaagcaagg ttttcagtat aatgttacat gcgtacacgc gtctgtacag
     5341 aaaaaaaaga aaaatttgaa atataaataa cgttcttaat actaacataa ctataaaaaa
     5401 ataaataggg acctagactt caggttgtct aactccttcc ttttcggtta gagcggatgt
     5461 ggggggaggg cgtgaatgta agcgtgacat aactaattac atgactcgag gtcgacggta
     5521 tcgataagct tgatatcgaa ttcctgcagc ccatcaccac tttgtacaag aaagctgggt
     5581 tttacattcc ccagccagaa gacttagaat ccatgcttga gccaaagcct ccattaaaac
     5641 cactgcccga ccctgcattg gatgctgatc cccaaccaat tgctgcacca gaattagagc
     5701 cactataaga gttatttcca gaaccgaagg cctggtttgg ctccctctgc atgttgcctt
     5761 ggttttggtt attacccgat gggcctgact ggttctgctg gctggctaac atgcccatca
     5821 taccccaact gctctttagt gctgcctggg cggcagccat catggctgga ttaatgctga
     5881 acgcaccaaa gttcatccca ccacccatat tactaccttg attgtttccc aaaccagctc
     5941 caccccctct gctattacca aatccaccct gattcccaaa gccacctgga ttaccaccaa
     6001 atcttccact tctttctaac tgtctattgc tattgtgctt aggttcggca ttggatatat
     6061 gaacgctgat tcctttaatg atcaagtcct ctccacaaag agactgcgca atctgatcat
     6121 ctgcaaatgt aacaaaggca aaggccctga atggcttggg gatgaagaca tccatcacat
     6181 ccccgtactg agagaagaac tcccgcagct catcctcagt catgtcctct gtacagcgcc
     6241 ccacaaacac ttttctgctt ctcaaaggct catcttggct ttgcttagaa ttaggaagtt
     6301 tgcagtcaca ccatcgtcca tctatcatat gtcgctgtga cattactttc acttgtgttt
     6361 catattccgt aaaacgaaca aagccaaacc cctttgaatg accagtctta agatctttct
     6421 tgacctgcac cataagaact tctccaaagg tactaaaata ctctttcagg tcctgttcgg
     6481 ttgttttcca tgggagaccc aacactatta aatcggatgt tttctggact gctcttttca
     6541 ctttcactgc tgatgaagca tctgtctcat ccatttttct tttgttatct tttggatagt
     6601 tgacaacata caccagattt ccccagccag catctggggc atgcagaatt ccttctacca
     6661 gccggacacc tctcatacac tgagacactg gattcctgta gcgaagccca cacgcccctg
     6721 gaaactgggc tgtaaccgtg gagagcagca ccgtcccatc gtcttccgat ggtatttcaa
     6781 tgggctcatc gttctcatct tcggttaccc gaatatattc agacattttg tagcctgctt
     6841 ttttgtacaa acttgtgatg ggggatccac tagttctag
//
