LOCUS       sequence-430961-        6879 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Galactose inducible expression of TDP-43-N267S.
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  JOURNAL   unpublished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            "unpublished"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-N267S
FEATURES             Location/Qualifiers
     source          1..6879
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(9..27)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     misc_feature    68..571
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     promoter        608..712
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(713..781,782..1573)
                     /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     complement(931..950)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1744..2332
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2233..2252
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2486..2503
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    2620..2641
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(2656..2661,2662..2679,2680..2686)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2694..2710
                     /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)."
     primer_bind     2699..2721
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     2718..2734
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2718..2734
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     2753..2773
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        2755..2773
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        2797..3238
                     /gene="S. cerevisiae GAL1"
                     /label=GAL1 promoter
                     /note="inducible promoter, regulated by Gal4"
     protein_bind    2797..2914
                     /label=UAS
                     /bound_moiety="Gal4"
                     /note="upstream activating sequence mediating 
                     Gal4-dependent induction"
     primer_bind     3209..3232
                     /label=GAL1
                     /note="S. cerevisiae GAL1 promoter, forward primer"
     primer_bind     3253..3269
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3253..3269
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     protein_bind    3277..3301
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(4553..4577)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(4610..4626)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4611..4627)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     terminator      4629..4876
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(4645..4663)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     primer_bind     complement(4894..4913)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4895..4913)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4923..4940)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4923..4939)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4932..4954)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      5080..5535
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5167..5186)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5377..5398
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(5666..6469)
                     /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"
     promoter        complement(6470..6685)
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     primer_bind     complement(6709..6728)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6828..6850
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
ORIGIN
        1 acgaaagggc ctcgtgatac gcctattttt ataggttaat gtcatgataa taatggtttc
       61 ttagacggat cgcttgcctg taacttacac gcgcctcgta tcttttaatg atggaataat
      121 ttgggaattt actctgtgtt tatttatttt tatgttttgt atttggattt tagaaagtaa
      181 ataaagaagg tagaagagtt acggaatgaa gaaaaaaaaa taaacaaagg tttaaaaaat
      241 ttcaacaaaa agcgtacttt acatatatat ttattagaca agaaaagcag attaaataga
      301 tatacattcg attaacgata agtaaaatgt aaaatcacag gattttcgtg tgtggtcttc
      361 tacacagaca agatgaaaca attcggcatt aatacctgag agcaggaaga gcaagataaa
      421 aggtagtatt tgttggcgat ccccctagag tcttttacat cttcggaaaa caaaaactat
      481 tttttcttta atttcttttt ttactttcta tttttaattt atatatttat attaaaaaat
      541 ttaaattata attattttta tagcacgtga tgaaaaggac ccaggtggca cttttcgggg
      601 aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct
      661 catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga gtatgagtat
      721 tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc ctgtttttgc
      781 tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg cacgagtggg
      841 ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc ccgaagaacg
      901 ttttccaatg atgagcactt ttaaagttct gctatgtggc gcggtattat cccgtattga
      961 cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact tggttgagta
     1021 ctcaccagtc acagaaaagc atcttacgga tggcatgaca gtaagagaat tatgcagtgc
     1081 tgccataacc atgagtgata acactgcggc caacttactt ctgacaacga tcggaggacc
     1141 gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc ttgatcgttg
     1201 ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt gacaccacga tgcctgtagc
     1261 aatggcaaca acgttgcgca aactattaac tggcgaacta cttactctag cttcccggca
     1321 acaattaata gactggatgg aggcggataa agttgcagga ccacttctgc gctcggccct
     1381 tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtgggt ctcgcggtat
     1441 cattgcagca ctggggccag atggtaagcc ctcccgtatc gtagttatct acacgacggg
     1501 gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg cctcactgat
     1561 taagcattgg taactgtcag accaagttta ctcatatata ctttagattg atttaaaact
     1621 tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat
     1681 cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc
     1741 ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct
     1801 accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg
     1861 cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt taggccacca
     1921 cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc
     1981 tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga
     2041 taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac
     2101 gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga
     2161 agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag
     2221 ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg
     2281 acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag
     2341 caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc
     2401 tgcgttatcc cctgattctg tggataaccg tattaccgcc tttgagtgag ctgataccgc
     2461 tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg aagagcgccc
     2521 aatacgcaaa ccgcctctcc ccgcgcgttg gccgattcat taatgcagct ggcacgacag
     2581 gtttcccgac tggaaagcgg gcagtgagcg caacgcaatt aatgtgagtt agctcactca
     2641 ttaggcaccc caggctttac actttatgct tccggctcgt atgttgtgtg gaattgtgag
     2701 cggataacaa tttcacacag gaaacagcta tgaccatgat tacgccaagc gcgcaattaa
     2761 ccctcactaa agggaacaaa agctggagct ctagtacgga ttagaagccg ccgagcgggt
     2821 gacagccctc cgaaggaaga ctctcctccg tgcgtcctcg tcttcaccgg tcgcgttcct
     2881 gaaacgcaga tgtgcctcgc gccgcactgc tccgaacaat aaagattcta caatactagc
     2941 ttttatggtt atgaagagga aaaattggca gtaacctggc cccacaaacc ttcaaatgaa
     3001 cgaatcaaat taacaaccat aggatgataa tgcgattagt tttttagcct tatttctggg
     3061 gtaattaatc agcgaagcga tgatttttga tctattaaca gatatataaa tgcaaaaact
     3121 gcataaccac tttaactaat actttcaaca ttttcggttt gtattacttc ttattcaaat
     3181 gtaataaaag tatcaacaaa aaattgttaa tatacctcta tactttaacg tcaaggagaa
     3241 aaaaccccgg attctagaac tagtggatcc cccatcacaa gtttgtacaa aaaagcaggc
     3301 tacaaaatgt ctgaatatat tcgggtaacc gaagatgaga acgatgagcc cattgaaata
     3361 ccatcggaag acgatgggac ggtgctgctc tccacggtta cagcccagtt tccaggggcg
     3421 tgtgggcttc gctacaggaa tccagtgtct cagtgtatga gaggtgtccg gctggtagaa
     3481 ggaattctgc atgccccaga tgctggctgg ggaaatctgg tgtatgttgt caactatcca
     3541 aaagataaca aaagaaaaat ggatgagaca gatgcttcat cagcagtgaa agtgaaaaga
     3601 gcagtccaga aaacatccga tttaatagtg ttgggtctcc catggaaaac aaccgaacag
     3661 gacctgaaag agtattttag tacctttgga gaagttctta tggtgcaggt caagaaagat
     3721 cttaagactg gtcattcaaa ggggtttggc tttgttcgtt ttacggaata tgaaacacaa
     3781 gtgaaagtaa tgtcacagcg acatatgata gatggacgat ggtgtgactg caaacttcct
     3841 aattctaagc aaagccaaga tgagcctttg agaagcagaa aagtgtttgt ggggcgctgt
     3901 acagaggaca tgactgagga tgagctgcgg gagttcttct ctcagtacgg ggatgtgatg
     3961 gatgtcttca tccccaagcc attcagggcc tttgcctttg ttacatttgc agatgatcag
     4021 attgcgcagt ctctttgtgg agaggacttg atcattaaag gaatcagcgt tcatatatcc
     4081 aatgccgaac ctaagcacaa tagcagtaga cagttagaaa gaagtggaag atttggtggt
     4141 aatccaggtg gctttgggaa tcagggtgga tttggtaata gcagaggggg tggagctggt
     4201 ttgggaaaca atcaaggtag taatatgggt ggtgggatga actttggtgc gttcagcatt
     4261 aatccagcca tgatggctgc cgcccaggca gcactacaga gcagttgggg tatgatgggc
     4321 atgttagcca gccagcagaa ccagtcaggc ccatcgggta ataaccaaaa ccaaggcaac
     4381 atgcagaggg agccaaacca ggccttcggt tctggaaata actcttatag tggctctaat
     4441 tctggtgcag caattggttg gggatcagca tccaatgcag ggtcgggcag tggttttaat
     4501 ggaggctttg gctcaagcat ggattctaag tcttctggct ggggaatgta aaacccagct
     4561 ttcttgtaca aagtggtgat gggctgcagg aattcgatat caagcttatc gataccgtcg
     4621 acctcgagtc atgtaattag ttatgtcacg cttacattca cgccctcccc ccacatccgc
     4681 tctaaccgaa aaggaaggag ttagacaacc tgaagtctag gtccctattt atttttttat
     4741 agttatgtta gtattaagaa cgttatttat atttcaaatt tttctttttt ttctgtacag
     4801 acgcgtgtac gcatgtaaca ttatactgaa aaccttgctt gagaaggttt tgggacgctc
     4861 gaaggcttta atttgcggcc ggtacccaat tcgccctata gtgagtcgta ttacgcgcgc
     4921 tcactggccg tcgttttaca acgtcgtgac tgggaaaacc ctggcgttac ccaacttaat
     4981 cgccttgcag cacatccccc tttcgccagc tggcgtaata gcgaagaggc ccgcaccgat
     5041 cgcccttccc aacagttgcg cagcctgaat ggcgaatgga cgcgccctgt agcggcgcat
     5101 taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccctag
     5161 cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc
     5221 aagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg cacctcgacc
     5281 ccaaaaaact tgattagggt gatggttcac gtagtgggcc atcgccctga tagacggttt
     5341 ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc caaactggaa
     5401 caacactcaa ccctatctcg gtctattctt ttgatttata agggattttg ccgatttcgg
     5461 cctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaatttt aacaaaatat
     5521 taacgcttac aatttcctga tgcggtattt tctccttacg catctgtgcg gtatttcaca
     5581 ccgcataggg taataactga tataattaaa ttgaagctct aatttgtgag tttagtatac
     5641 atgcatttac ttataataca gttttttagt tttgctggcc gcatcttctc aaatatgctt
     5701 cccagcctgc ttttctgtaa cgttcaccct ctaccttagc atcccttccc tttgcaaata
     5761 gtcctcttcc aacaataata atgtcagatc ctgtagagac cacatcatcc acggttctat
     5821 actgttgacc caatgcgtct cccttgtcat ctaaacccac accgggtgtc ataatcaacc
     5881 aatcgtaacc ttcatctctt ccacccatgt ctctttgagc aataaagccg ataacaaaat
     5941 ctttgtcgct cttcgcaatg tcaacagtac ccttagtata ttctccagta gatagggagc
     6001 ccttgcatga caattctgct aacatcaaaa ggcctctagg ttcctttgtt acttcttctg
     6061 ccgcctgctt caaaccgcta acaatacctg ggcccaccac accgtgtgca ttcgtaatgt
     6121 ctgcccattc tgctattctg tatacacccg cagagtactg caatttgact gtattaccaa
     6181 tgtcagcaaa ttttctgtct tcgaagagta aaaaattgta cttggcggat aatgccttta
     6241 gcggcttaac tgtgccctcc atggaaaaat cagtcaagat atccacatgt gtttttagta
     6301 aacaaatttt gggacctaat gcttcaacta actccagtaa ttccttggtg gtacgaacat
     6361 ccaatgaagc acacaagttt gtttgctttt cgtgcatgat attaaatagc ttggcagcaa
     6421 caggactagg atgagtagca gcacgttcct tatatgtagc tttcgacatg atttatcttc
     6481 gtttcctgca ggtttttgtt ctgtgcagtt gggttaagaa tactgggcaa tttcatgttt
     6541 cttcaacact acatatgcgt atatatacca atctaagtct gtgctccttc cttcgttctt
     6601 ccttctgttc ggagattacc gaatcaaaaa aatttcaagg aaaccgaaat caaaaaaaag
     6661 aataaaaaaa aaatgatgaa ttgaaaaggt ggtatggtgc actctcagta caatctgctc
     6721 tgatgccgca tagttaagcc agccccgaca cccgccaaca cccgctgacg cgccctgacg
     6781 ggcttgtctg ctcccggcat ccgcttacag acaagctgtg accgtctccg ggagctgcat
     6841 gtgtcagagg ttttcaccgt catcaccgaa acgcgcgag
//
