LOCUS       sequence-439443-        6213 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Galactose inducible expression of Alpha-synuclein-A53T.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6213)
  AUTHORS   Chavez Lab
  TITLE     Multiplex screening identifies broad modifiers of ALS-associated 
            proteotoxicity
  JOURNAL   unpublished
REFERENCE   2  (bases 1 to 6213)
  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            "unpublished"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-Alpha-synuclein-A53T
FEATURES             Location/Qualifiers
     source          1..6213
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(join(251..1042,1043..1111))
                     /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     874..893
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1112..1216)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      1243..2585
                     /label=2u ori
                     /note="yeast 2u plasmid origin of replication"
     protein_bind    complement(2206..2253)
                     /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)."
     primer_bind     2626..2644
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(2682..2704)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     2804..2823
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        2847..3062
                     /gene="S. cerevisiae URA3"
                     /label=URA3 promoter
     CDS             3063..3866
                     /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(3997..4452)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4134..4155)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4346..4365
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4578..4600
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     4592..4609
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     4593..4609
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4619..4638
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        4619..4637
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     terminator      complement(4656..4903)
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     4869..4887
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     primer_bind     4905..4921
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     4906..4922
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    4955..4979
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(5409..5433)
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(5441..5457)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5441..5457)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     promoter        complement(5472..5913)
                     /gene="S. cerevisiae GAL1"
                     /label=GAL1 promoter
                     /note="inducible promoter, regulated by Gal4"
     primer_bind     complement(5478..5501)
                     /label=GAL1
                     /note="S. cerevisiae GAL1 promoter, forward primer"
     protein_bind    5796..5913
                     /label=UAS
                     /bound_moiety="Gal4"
                     /note="upstream activating sequence mediating 
                     Gal4-dependent induction"
     primer_bind     complement(5937..5957)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(5937..5955)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(5976..5992)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5976..5992)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5989..6011)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6000..6016
                     /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(6024..6030,6031..6048,6049..6054))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6069..6090
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
ORIGIN
        1 ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc
       61 gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt
      121 ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct
      181 agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt
      241 ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc
      301 gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac
      361 catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat
      421 cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg
      481 cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata
      541 gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta
      601 tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt
      661 gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag
      721 tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa
      781 gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc
      841 gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt
      901 taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc
      961 tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta
     1021 ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa
     1081 taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca
     1141 tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac
     1201 aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgaacgaagc atctgtgctt
     1261 cattttgtag aacaaaaatg caacgcgaga gcgctaattt ttcaaacaaa gaatctgagc
     1321 tgcattttta cagaacagaa atgcaacgcg aaagcgctat tttaccaacg aagaatctgt
     1381 gcttcatttt tgtaaaacaa aaatgcaacg cgagagcgct aatttttcaa acaaagaatc
     1441 tgagctgcat ttttacagaa cagaaatgca acgcgagagc gctattttac caacaaagaa
     1501 tctatacttc ttttttgttc tacaaaaatg catcccgaga gcgctatttt tctaacaaag
     1561 catcttagat tacttttttt ctcctttgtg cgctctataa tgcagtctct tgataacttt
     1621 ttgcactgta ggtccgttaa ggttagaaga aggctacttt ggtgtctatt ttctcttcca
     1681 taaaaaaagc ctgactccac ttcccgcgtt tactgattac tagcgaagct gcgggtgcat
     1741 tttttcaaga taaaggcatc cccgattata ttctataccg atgtggattg cgcatacttt
     1801 gtgaacagaa agtgatagcg ttgatgattc ttcattggtc agaaaattat gaacggtttc
     1861 ttctattttg tctctatata ctacgtatag gaaatgttta cattttcgta ttgttttcga
     1921 ttcactctat gaatagttct tactacaatt tttttgtcta aagagtaata ctagagataa
     1981 acataaaaaa tgtagaggtc gagtttagat gcaagttcaa ggagcgaaag gtggatgggt
     2041 aggttatata gggatatagc acagagatat atagcaaaga gatacttttg agcaatgttt
     2101 gtggaagcgg tattcgcaat attttagtag ctcgttacag tccggtgcgt ttttggtttt
     2161 ttgaaagtgc gtcttcagag cgcttttggt tttcaaaagc gctctgaagt tcctatactt
     2221 tctagagaat aggaacttcg gaataggaac ttcaaagcgt ttccgaaaac gagcgcttcc
     2281 gaaaatgcaa cgcgagctgc gcacatacag ctcactgttc acgtcgcacc tatatctgcg
     2341 tgttgcctgt atatatatat acatgagaag aacggcatag tgcgtgttta tgcttaaatg
     2401 cgtacttata tgcgtctatt tatgtaggat gaaaggtagt ctagtacctc ctgtgatatt
     2461 atcccattcc atgcggggta tcgtatgctt ccttcagcac taccctttag ctgttctata
     2521 tgctgccact cctcaattgg attagtctca tccttcaatg ctatcatttc ctttgatatt
     2581 ggatcatact aagaaaccat tattatcatg acattaacct ataaaaatag gcgtatcacg
     2641 aggccctttc gtctcgcgcg tttcggtgat gacggtgaaa acctctgaca catgcagctc
     2701 ccggagacgg tcacagcttg tctgtaagcg gatgccggga gcagacaagc ccgtcagggc
     2761 gcgtcagcgg gtgttggcgg gtgtcggggc tggcttaact atgcggcatc agagcagatt
     2821 gtactgagag tgcaccatac caccttttca attcatcatt ttttttttat tctttttttt
     2881 gatttcggtt tccttgaaat ttttttgatt cggtaatctc cgaacagaag gaagaacgaa
     2941 ggaaggagca cagacttaga ttggtatata tacgcatatg tagtgttgaa gaaacatgaa
     3001 attgcccagt attcttaacc caactgcaca gaacaaaaac ctgcaggaaa cgaagataaa
     3061 tcatgtcgaa agctacatat aaggaacgtg ctgctactca tcctagtcct gttgctgcca
     3121 agctatttaa tatcatgcac gaaaagcaaa caaacttgtg tgcttcattg gatgttcgta
     3181 ccaccaagga attactggag ttagttgaag cattaggtcc caaaatttgt ttactaaaaa
     3241 cacatgtgga tatcttgact gatttttcca tggagggcac agttaagccg ctaaaggcat
     3301 tatccgccaa gtacaatttt ttactcttcg aagacagaaa atttgctgac attggtaata
     3361 cagtcaaatt gcagtactct gcgggtgtat acagaatagc agaatgggca gacattacga
     3421 atgcacacgg tgtggtgggc ccaggtattg ttagcggttt gaagcaggcg gcagaagaag
     3481 taacaaagga acctagaggc cttttgatgt tagcagaatt gtcatgcaag ggctccctat
     3541 ctactggaga atatactaag ggtactgttg acattgcgaa gagcgacaaa gattttgtta
     3601 tcggctttat tgctcaaaga gacatgggtg gaagagatga aggttacgat tggttgatta
     3661 tgacacccgg tgtgggttta gatgacaagg gagacgcatt gggtcaacag tatagaaccg
     3721 tggatgatgt ggtctctaca ggatctgaca ttattattgt tggaagagga ctatttgcaa
     3781 agggaaggga tgctaaggta gagggtgaac gttacagaaa agcaggctgg gaagcatatt
     3841 tgagaagatg cggccagcaa aactaaaaaa ctgtattata agtaaatgca tgtatactaa
     3901 actcacaaat tagagcttca atttaattat atcagttatt accctatgcg gtgtgaaata
     3961 ccgcacagat gcgtaaggag aaaataccgc atcaggaaat tgtaagcgtt aatattttgt
     4021 taaaattcgc gttaaatttt tgttaaatca gctcattttt taaccaatag gccgaaatcg
     4081 gcaaaatccc ttataaatca aaagaataga ccgagatagg gttgagtgtt gttccagttt
     4141 ggaacaagag tccactatta aagaacgtgg actccaacgt caaagggcga aaaaccgtct
     4201 atcagggcga tggcccacta cgtgaaccat caccctaatc aagttttttg gggtcgaggt
     4261 gccgtaaagc actaaatcgg aaccctaaag ggagcccccg atttagagct tgacggggaa
     4321 agccggcgaa cgtggcgaga aaggaaggga agaaagcgaa aggagcgggc gctagggcgc
     4381 tggcaagtgt agcggtcacg ctgcgcgtaa ccaccacacc cgccgcgctt aatgcgccgc
     4441 tacagggcgc gtccattcgc cattcaggct gcgcaactgt tgggaagggc gatcggtgcg
     4501 ggcctcttcg ctattacgcc agctggcgaa agggggatgt gctgcaaggc gattaagttg
     4561 ggtaacgcca gggttttccc agtcacgacg ttgtaaaacg acggccagtg agcgcgcgta
     4621 atacgactca ctatagggcg aattgggtac cggccgcaaa ttaaagcctt cgagcgtccc
     4681 aaaaccttct caagcaaggt tttcagtata atgttacatg cgtacacgcg tctgtacaga
     4741 aaaaaaagaa aaatttgaaa tataaataac gttcttaata ctaacataac tataaaaaaa
     4801 taaataggga cctagacttc aggttgtcta actccttcct tttcggttag agcggatgtg
     4861 gggggagggc gtgaatgtaa gcgtgacata actaattaca tgactcgagg tcgacggtat
     4921 cgataagctt gatatcgaat tcctgcagcc catcaccact ttgtacaaga aagctgggtt
     4981 ttaggcttca ggttcgtagt cttgataccc ttcctcagaa ggcatttcat aagcctcatt
     5041 gtcaggatcc acaggcatat cttccagaat tccttcctgt ggggctcctt cttcattctt
     5101 gcccaactgg tcctttttga caaagccagt ggctgctgca atgctccctg ctccctccac
     5161 tgtcttctgg gctactgctg tcacacccgt caccactgct cctccaacat ttgtcacttg
     5221 ctctttggtc ttctcagcca ctgttgtcac accatgcacc actccctcct tggttttgga
     5281 gcctacatag agaacaccct cttttgtctt tcctgctgct tctgccacac cctgtttggt
     5341 tttctcagca gcagccacaa ctccctcctt ggcctttgaa agtcctttca tgaatacatc
     5401 cattttgtag cctgcttttt tgtacaaact tgtgatgggg gatccactag ttctagaatc
     5461 cggggttttt tctccttgac gttaaagtat agaggtatat taacaatttt ttgttgatac
     5521 ttttattaca tttgaataag aagtaataca aaccgaaaat gttgaaagta ttagttaaag
     5581 tggttatgca gtttttgcat ttatatatct gttaatagat caaaaatcat cgcttcgctg
     5641 attaattacc ccagaaataa ggctaaaaaa ctaatcgcat tatcatccta tggttgttaa
     5701 tttgattcgt tcatttgaag gtttgtgggg ccaggttact gccaattttt cctcttcata
     5761 accataaaag ctagtattgt agaatcttta ttgttcggag cagtgcggcg cgaggcacat
     5821 ctgcgtttca ggaacgcgac cggtgaagac gaggacgcac ggaggagagt cttccttcgg
     5881 agggctgtca cccgctcggc ggcttctaat ccgtactaga gctccagctt ttgttccctt
     5941 tagtgagggt taattgcgcg cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat
     6001 tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg taaagcctgg
     6061 ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc cgctttccag
     6121 tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt
     6181 ttgcgtattg ggcgctcttc cgcttcctcg ctc
//
