LOCUS       sequence-167362-        5798 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  induce expression of dipeptide repeat GR in yeast.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5798)
  AUTHORS   Jovicic A, Mertens J, Boeynaems S, Bogaert E, Chai N, Yamada SB, 
            Paul JW 3rd, Sun S, Herdy JR, Bieri G, Kramer NJ, Gage FH, Van Den 
            Bosch L, Robberecht W, Gitler AD
  TITLE     Modifiers of C9orf72 dipeptide repeat toxicity connect 
            nucleocytoplasmic transport defects to FTD/ALS.
  JOURNAL   Nat Neurosci. 2015 Sep;18(9):1226-9. doi: 10.1038/nn.4085.
  PUBMED    26308983
REFERENCE   2  (bases 1 to 5798)
  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"; doi: "10.1038/nn"; 
            journalName: "Nat Neurosci"; date: "2015-09"; volume: "18"; issue: 
            "9"; pages: "1226-9"
COMMENT     Sequence Label: pAG303-Gal-GR100
FEATURES             Location/Qualifiers
     source          1..5798
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(29..51)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     151..170
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        315..502
                     /gene="S. cerevisiae HIS3"
                     /label=HIS3 promoter
     CDS             503..1162
                     /codon_start=1
                     /gene="S. cerevisiae HIS3"
                     /product="imidazoleglycerol-phosphate dehydratase, required
                     for histidine biosynthesis"
                     /label=HIS3
                     /note="yeast auxotrophic marker"
                     /translation="MTEQKALVKRITNETKIQIAISLKGGPLAIEHSIFPEKEAEAVAE
                     QATQSQVINVHTGIGFLDHMIHALAKHSGWSLIVECIGDLHIDDHHTTEDCGIALGQAF
                     KEALLARGVKRFGSGFAPLDEALSRAVVDLSNRPYAVVELGLQREKVGDLSCEMIPHFL
                     ESFAEASRITLHVDCLRGKNDHHRSESAFKALAVAIREATSPNGTNDVPSTKGVLM"
     rep_origin      complement(1423..1878)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1560..1581)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     1772..1791
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2008..2030
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     2022..2039
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     2023..2039
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2046..2065
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2046..2064
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        2084..2525
                     /gene="S. cerevisiae GAL1"
                     /label=GAL1 promoter
                     /note="inducible promoter, regulated by Gal4"
     protein_bind    2084..2201
                     /label=UAS
                     /bound_moiety="Gal4"
                     /note="upstream activating sequence mediating 
                     Gal4-dependent induction"
     primer_bind     2496..2519
                     /label=GAL1
                     /note="S. cerevisiae GAL1 promoter, forward primer"
     primer_bind     2540..2556
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2540..2556
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     protein_bind    2564..2588
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(3202..3226)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(3259..3275)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3260..3276)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     terminator      3278..3525
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(3294..3312)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     promoter        complement(3548..3566)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(3587..3603)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3587..3603)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3600..3622)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3611..3627
                     /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(3635..3641,3642..3659,3660..3665))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     primer_bind     complement(3818..3835)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3989..4577)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(join(4748..5539,5540..5608))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFFHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     5371..5390
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5609..5713)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5771..5789
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca
       61 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg
      121 ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc
      181 accataattc cgttttaaga gcttggtgag cgctaggagt cactgccagg tatcgtttga
      241 acacggcatt agtcagggaa gtcataacac agtcctttcc cgcaattttc tttttctatt
      301 actcttggcc tcctctagta cactctatat ttttttatgc ctcggtaatg attttcattt
      361 ttttttttcc acctagcgga tgactctttt tttttcttag cgattggcat tatcacataa
      421 tgaattatac attatataaa gtaatgtgat ttcttcgaag aatatactaa aaaatgagca
      481 ggcaagataa acgaaggcaa agatgacaga gcagaaagcc ctagtaaagc gtattacaaa
      541 tgaaaccaag attcagattg cgatctcttt aaagggtggt cccctagcga tagagcactc
      601 gatcttccca gaaaaagagg cagaagcagt agcagaacag gccacacaat cgcaagtgat
      661 taacgtccac acaggtatag ggtttctgga ccatatgata catgctctgg ccaagcattc
      721 cggctggtcg ctaatcgttg agtgcattgg tgacttacac atagacgacc atcacaccac
      781 tgaagactgc gggattgctc tcggtcaagc ttttaaagag gccctactgg cgcgtggagt
      841 aaaaaggttt ggatcaggat ttgcgccttt ggatgaggca ctttccagag cggtggtaga
      901 tctttcgaac aggccgtacg cagttgtcga acttggtttg caaagggaga aagtaggaga
      961 tctctcttgc gagatgatcc cgcattttct tgaaagcttt gcagaggcta gcagaattac
     1021 cctccacgtt gattgtctgc gaggcaagaa tgatcatcac cgtagtgaga gtgcgttcaa
     1081 ggctcttgcg gttgccataa gagaagccac ctcgcccaat ggtaccaacg atgttccctc
     1141 caccaaaggt gttcttatgt agtgacaccg attatttaaa gctgcagcat acgatatata
     1201 tacatgtgta tatatgtata cctatgaatg tcagtaagta tgtatacgaa cagtatgata
     1261 ctgaagatga caaggtaatg catcattcta tacgtgtcat tctgaacgag gcgcgctttc
     1321 cttttttctt tttgcttttt cttttttttt ctcttgaact cgacggatca tatgcggtgt
     1381 gaaataccgc acagatgcgt aaggagaaaa taccgcatca ggaaattgta aacgttaata
     1441 ttttgttaaa attcgcgtta aatttttgtt aaatcagctc attttttaac caataggccg
     1501 aaatcggcaa aatcccttat aaatcaaaag aatagaccga gatagggttg agtgttgttc
     1561 cagtttggaa caagagtcca ctattaaaga acgtggactc caacgtcaaa gggcgaaaaa
     1621 ccgtctatca gggcgatggc ccactacgtg aaccatcacc ctaatcaagt tttttggggt
     1681 cgaggtgccg taaagcacta aatcggaacc ctaaagggag cccccgattt agagcttgac
     1741 ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa agcgaaagga gcgggcgcta
     1801 gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac cacacccgcc gcgcttaatg
     1861 cgccgctaca gggcgcgtcg cgccattcgc cattcaggct gcgcaactgt tgggaagggc
     1921 gatcggtgcg ggcctcttcg ctattacgcc agctggcgaa ggggggatgt gctgcaaggc
     1981 gattaagttg ggtaacgcca gggttttccc agtcacgacg ttgtaaaacg acggccagtg
     2041 aattgtaata cgactcacta tagggcgaat tggagctcta gtacggatta gaagccgccg
     2101 agcgggtgac agccctccga aggaagactc tcctccgtgc gtcctcgtct tcaccggtcg
     2161 cgttcctgaa acgcagatgt gcctcgcgcc gcactgctcc gaacaataaa gattctacaa
     2221 tactagcttt tatggttatg aagaggaaaa attggcagta acctggcccc acaaaccttc
     2281 aaatgaacga atcaaattaa caaccatagg atgataatgc gattagtttt ttagccttat
     2341 ttctggggta attaatcagc gaagcgatga tttttgatct attaacagat atataaatgc
     2401 aaaaactgca taaccacttt aactaatact ttcaacattt tcggtttgta ttacttctta
     2461 ttcaaatgta ataaaagtat caacaaaaaa ttgttaatat acctctatac tttaacgtca
     2521 aggagaaaaa accccggatt ctagaactag tggatccccc atcacaagtt tgtacaaaaa
     2581 agcaggcttc aaaaatgggt agaggtagag gtagaggtag aggtagaggt agaggtagag
     2641 gtagaggtag aggtagaggt agaggtagag gtagaggtag aggtagaggt agaggtagag
     2701 gtagaggtag aggtagaggt agaggtagag gtagaggtag aggtagaggt agaggtagag
     2761 gtagaggtag aggtagaggt agaggtagag gtagaggtag aggtagaggt agaggtagag
     2821 gtagaggtag aggtagaggt agaggtagag gtagaggtag aggtagaggt agaggtagag
     2881 gtagaggtag aggtagaggt agaggtagag gtagaggtag aggtagaggt agaggtagag
     2941 gtagaggtag aggtagaggt agaggtagag gtagaggtag aggtagaggt agaggtagag
     3001 gtagaggtag aggtagaggt agaggtagag gtagaggtag aggtagaggt agaggtagag
     3061 gtagaggtag aggtagaggt agaggtagag gtagaggtag aggtagaggt agaggtagag
     3121 gtagaggtag aggtagaggt agaggtagag gtagaggtag aggtagaggt agaggtagag
     3181 gtagaggtag aggtagataa gacccagctt tcttgtacaa agtggtgatg ggctgcagga
     3241 attcgatatc aagcttatcg ataccgtcga cctcgagtca tgtaattagt tatgtcacgc
     3301 ttacattcac gccctccccc cacatccgct ctaaccgaaa aggaaggagt tagacaacct
     3361 gaagtctagg tccctattta tttttttata gttatgttag tattaagaac gttatttata
     3421 tttcaaattt ttcttttttt tctgtacaga cgcgtgtacg catgtaacat tatactgaaa
     3481 accttgcttg agaaggtttt gggacgctcg aaggctttaa tttgcggccg gtacccagct
     3541 tttgttccct ttagtgaggg ttaattccga gcttggcgta atcatggtca tagctgtttc
     3601 ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat aggagccgga agcataaagt
     3661 gtaaagcctg gggtgcctaa tgagtgaggt aactcacatt aattgcgttg cgctcactgc
     3721 ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg
     3781 ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct
     3841 cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca
     3901 cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga
     3961 accgtaaaaa ggccgcgttg ctggcgtttt tccataggct cggcccccct gacgagcatc
     4021 acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg
     4081 cgttcccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat
     4141 acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcaatgctca cgctgtaggt
     4201 atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc
     4261 agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg
     4321 acttatcgcc actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg
     4381 gtgctacaga gttcttgaag tggtggccta actacggcta cactagaagg acagtatttg
     4441 gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg
     4501 gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca
     4561 gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga
     4621 acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga
     4681 tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt
     4741 ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt
     4801 catccatagt tgcctgactg cccgtcgtgt agataactac gatacgggag ggcttaccat
     4861 ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag
     4921 caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct
     4981 ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt
     5041 tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg
     5101 cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgaa
     5161 aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt
     5221 tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat
     5281 gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac
     5341 cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa
     5401 aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt
     5461 tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt
     5521 tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa
     5581 gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt
     5641 atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa
     5701 taggggttcc gcgcacattt ccccgaaaag tgccacctga cgtcttatta tcatgacatt
     5761 aacctataaa aataggcgta tcacgaggcc ctttcgtc
//
