LOCUS       sequence-261213-        7666 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Httex1 (without proline-rich region) fused to ymsfGFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7666)
  AUTHORS   Albakri MB, Jiang Y, Genereaux J, Lajoie P
  TITLE     Polyglutamine toxicity assays highlight the advantages of mScarlet 
            for imaging in Saccharomyces cerevisiae.
  JOURNAL   F1000Res. 2018 Aug 10;7:1242. doi: 10.12688/f1000research.15829.2. 
            eCollection 2018.
  PUBMED    30631438
REFERENCE   2  (bases 1 to 7666)
  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: "F1000Res.";
            date: "2018-08-10"; volume: "7:1242. doi"; pages: " 
            10.12688/f1000research.15829.2. eCollection 2018"
COMMENT     Sequence Label: 68Q-ymsfGFP
FEATURES             Location/Qualifiers
     source          1..7666
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             27..50
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     terminator      1059..1306
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(1075..1093)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     primer_bind     complement(1324..1343)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1325..1343)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1353..1370)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1353..1369)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1362..1384)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      1510..1965
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1597..1616)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1807..1828
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        2266..2673
                     /gene="S. cerevisiae LEU2"
                     /label=LEU2 promoter
     CDS             2674..3768
                     /codon_start=1
                     /gene="S. cerevisiae LEU2"
                     /product="3-isopropylmalate dehydrogenase, required for 
                     leucine biosynthesis"
                     /label=LEU2
                     /note="yeast auxotrophic marker"
                     /translation="MSAPKKIVVLPGDHVGQEITAEAIKVLKAISDVRSNVKFDFENHL
                     IGGAAIDATGVPLPDEALEASKKADAVLLGAVGGPKWGTGSVRPEQGLLKIRKELQLYA
                     NLRPCNFASDSLLDLSPIKPQFAKGTDFVVVRELVGGIYFGKRKEDDGDGVAWDSEQYT
                     VPEVQRITRMAAFMALQHEPPLPIWSLDKANVLASSRLWRKTVEETIKNEFPTLKVQHQ
                     LIDSAAMILVKNPTHLNGIIITSNMFGDIISDEASVIPGSLGLLPSASLASLPDKNTAF
                     GLYEPCHGSAPDLPKNKVNPIATILSAAMMLKLSLNLPEEGKAIEDAVKKVLDAGIRTG
                     DLGGSNSTTEVGDAVAEEVKKILA"
     primer_bind     complement(4262..4281)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4381..4403
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4441..4459)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     misc_feature    4501..5004
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     promoter        5041..5145
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5146..5214,5215..6006)
                     /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(5364..5383)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6177..6765
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6666..6685
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6919..6936
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    7053..7074
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7089..7094,7095..7112,7113..7119)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7127..7143
                     /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     7132..7154
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7151..7167
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7151..7167
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     7186..7206
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        7188..7206
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        7225..7666
                     /gene="S. cerevisiae GAL1"
                     /label=GAL1 promoter
                     /note="inducible promoter, regulated by Gal4"
     protein_bind    7225..7342
                     /label=UAS
                     /bound_moiety="Gal4"
                     /note="upstream activating sequence mediating 
                     Gal4-dependent induction"
     primer_bind     7637..7660
                     /label=GAL1
                     /note="S. cerevisiae GAL1 promoter, forward primer"
ORIGIN
        1 aaaaaacccc ggattctaga gccatggact acaaggacga cgatgacaag ctggcgaccc
       61 tggaaaagct aatgaaggcc ttcgagtccc tcaaaagctt ccaacagcag caacagcaac
      121 aacagcagca acagcaacaa cagcagcaac agcaacaaca gcagcaacag caacaacagc
      181 agcaacagca acaacagcag cagcagcaac aacagcagca acagcaacaa caacagcagc
      241 aacagcaaca acagcagcaa cagcaacaac agcagcaaca gcaacaacag cagcaacagc
      301 aacaactgca gcccggggga tccactagtg tatccaaagg tgaagaattg tttactggtg
      361 ttgttcctat tttagttgaa ttagacggtg acgttaatgg tcataagttt agtgttagag
      421 gtgaaggtga aggtgacgca actaacggta aattgacatt gaagtttatt tgtactactg
      481 gtaaattgcc agttccatgg ccaaccttgg ttaccacttt aacttatggt gttcaatgct
      541 tctcaagata ccctgatcat atgaaaagac acgacttttt caagagtgcc atgccagaag
      601 gttacgtaca agaaagaact atatctttta aagatgacgg tacatacaag accagagctg
      661 aagtaaaatt cgaaggtgac acattggtca acagaatcga attgaagggt atcgatttca
      721 aggaagacgg taacatcttg ggtcataaat tagaatacaa tttcaactca cacaatgttt
      781 acatcactgc tgataagcaa aagaatggta tcaaagcaaa cttcaagatc agacataacg
      841 ttgaagatgg ttccgtacaa ttagctgacc actatcaaca aaatacacct attggtgacg
      901 gtcctgtttt gttaccagac aaccattact tgtctaccca atcaaaattg tccaaagatc
      961 caaacgaaaa gagagaccat atggtcttgt tggaatttgt cactgccgct ggtatcactc
     1021 acggtatgga tgaattgtat aagtaggtcg acctcgagtc atgtaattag ttatgtcacg
     1081 cttacattca cgccctcccc ccacatccgc tctaaccgaa aaggaaggag ttagacaacc
     1141 tgaagtctag gtccctattt atttttttat agttatgtta gtattaagaa cgttatttat
     1201 atttcaaatt tttctttttt ttctgtacag acgcgtgtac gcatgtaaca ttatactgaa
     1261 aaccttgctt gagaaggttt tgggacgctc gaaggcttta atttgcggcc ggtacccaat
     1321 tcgccctata gtgagtcgta ttacgcgcgc tcactggccg tcgttttaca acgtcgtgac
     1381 tgggaaaacc ctggcgttac ccaacttaat cgccttgcag cacatccccc tttcgccagc
     1441 tggcgtaata gcgaagaggc ccgcaccgat cgcccttccc aacagttgcg cagcctgaat
     1501 ggcgaatgga cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca
     1561 gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc tttcgctttc ttcccttcct
     1621 ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcgggggctc cctttagggt
     1681 tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgattagggt gatggttcac
     1741 gtagtgggcc atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct
     1801 ttaatagtgg actcttgttc caaactggaa caacactcaa ccctatctcg gtctattctt
     1861 ttgatttata agggattttg ccgatttcgg cctattggtt aaaaaatgag ctgatttaac
     1921 aaaaatttaa cgcgaatttt aacaaaatat taacgcttac aatttcctga tgcggtattt
     1981 tctccttacg catctgtgcg gtatttcaca ccgcatatcg accggtcgag gagaacttct
     2041 agtatatcta catacctaat attattgcct tattaaaaat ggaatcccaa caattacatc
     2101 aaaatccaca ttctcttcaa aatcaattgt cctgtacttc cttgttcatg tgtgttcaaa
     2161 aacgttatat ttataggata attatactct atttctcaac aagtaattgg ttgtttggcc
     2221 gagcggtcta aggcgcctga ttcaagaaat atcttgaccg cagttaactg tgggaatact
     2281 caggtatcgt aagatgcaag agttcgaatc tcttagcaac cattattttt ttcctcaaca
     2341 taacgagaac acacaggggc gctatcgcac agaatcaaat tcgatgactg gaaatttttt
     2401 gttaatttca gaggtcgcct gacgcatata cctttttcaa ctgaaaaatt gggagaaaaa
     2461 ggaaaggtga gagcgccgga accggctttt catatagaat agagaagcgt tcatgactaa
     2521 atgcttgcat cacaatactt gaagttgaca atattattta aggacctatt gttttttcca
     2581 ataggtggtt agcaatcgtc ttactttcta acttttctta ccttttacat ttcagcaata
     2641 tatatatata tatttcaagg atataccatt ctaatgtctg cccctaagaa gatcgtcgtt
     2701 ttgccaggtg accacgttgg tcaagaaatc acagccgaag ccattaaggt tcttaaagct
     2761 atttctgatg ttcgttccaa tgtcaagttc gatttcgaaa atcatttaat tggtggtgct
     2821 gctatcgatg ctacaggtgt tccacttcca gatgaggcgc tggaagcctc caagaaggct
     2881 gatgccgttt tgttaggtgc tgtgggtggt cctaaatggg gtaccggtag tgttagacct
     2941 gaacaaggtt tactaaaaat ccgtaaagaa cttcaattgt acgccaactt aagaccatgt
     3001 aactttgcat ccgactctct tttagactta tctccaatca agccacaatt tgctaaaggt
     3061 actgacttcg ttgttgtcag agaattagtg ggaggtattt actttggtaa gagaaaggaa
     3121 gacgatggtg atggtgtcgc ttgggatagt gaacaataca ccgttccaga agtgcaaaga
     3181 atcacaagaa tggccgcttt catggcccta caacatgagc caccattgcc tatttggtcc
     3241 ttggataaag ctaatgtttt ggcctcttca agattatgga gaaaaactgt ggaggaaacc
     3301 atcaagaacg aattccctac attgaaggtt caacatcaat tgattgattc tgccgccatg
     3361 atcctagtta agaacccaac ccacctaaat ggtattataa tcaccagcaa catgtttggt
     3421 gatatcatct ccgatgaagc ctccgttatc ccaggttcct tgggtttgtt gccatctgcg
     3481 tccttggcct ctttgccaga caagaacacc gcatttggtt tgtacgaacc atgccacggt
     3541 tctgctccag atttgccaaa gaataaggtc aaccctatcg ccactatctt gtctgctgca
     3601 atgatgttga aattgtcatt gaacttgcct gaagaaggta aggccattga agatgcagtt
     3661 aaaaaggttt tggatgcagg tatcagaact ggtgatttag gtggttccaa cagtaccacc
     3721 gaagtcggtg atgctgtcgc cgaagaagtt aagaaaatcc ttgcttaaaa agattctctt
     3781 tttttatgat atttgtacat aaactttata aatgaaattc ataatagaaa cgacacgaaa
     3841 ttacaaaatg gaatatgttc atagggtaga cgaaactata tacgcaatct acatacattt
     3901 atcaagaagg agaaaaagga ggatgtaaag gaatacaggt aagcaaattg atactaatgg
     3961 ctcaacgtga taaggaaaaa gaattgcact ttaacattaa tattgacaag gaggagggca
     4021 ccacacaaaa agttaggtgt aacagaaaat catgaaacta tgattcctaa tttatatatt
     4081 ggaggatttt ctctaaaaaa aaaaaaatac aacaaataaa aaacactcaa tgacctgacc
     4141 atttgatgga gtttaagtca ataccttctt gaaccatttc ccataatggt gaaagttccc
     4201 tcaagaattt tactctgtca gaaacggcct taacgacgta gtcgatatgg tgcactctca
     4261 gtacaatctg ctctgatgcc gcatagttaa gccagccccg acacccgcca acacccgctg
     4321 acgcgccctg acgggcttgt ctgctcccgg catccgctta cagacaagct gtgaccgtct
     4381 ccgggagctg catgtgtcag aggttttcac cgtcatcacc gaaacgcgcg agacgaaagg
     4441 gcctcgtgat acgcctattt ttataggtta atgtcatgat aataatggtt tcttaggacg
     4501 gatcgcttgc ctgtaactta cacgcgcctc gtatctttta atgatggaat aatttgggaa
     4561 tttactctgt gtttatttat ttttatgttt tgtatttgga ttttagaaag taaataaaga
     4621 aggtagaaga gttacggaat gaagaaaaaa aaataaacaa aggtttaaaa aatttcaaca
     4681 aaaagcgtac tttacatata tatttattag acaagaaaag cagattaaat agatatacat
     4741 tcgattaacg ataagtaaaa tgtaaaatca caggattttc gtgtgtggtc ttctacacag
     4801 acaagatgaa acaattcggc attaatacct gagagcagga agagcaagat aaaaggtagt
     4861 atttgttggc gatcccccta gagtctttta catcttcgga aaacaaaaac tattttttct
     4921 ttaatttctt tttttacttt ctatttttaa tttatatatt tatattaaaa aatttaaatt
     4981 ataattattt ttatagcacg tgatgaaaag gacccaggtg gcacttttcg gggaaatgtg
     5041 cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga
     5101 caataaccct gataaatgct tcaataatat tgaaaaagga agagtatgag tattcaacat
     5161 ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt tgctcaccca
     5221 gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt gggttacatc
     5281 gaactggatc tcaacagcgg taagatcctt gagagttttc gccccgaaga acgttttcca
     5341 atgatgagca cttttaaagt tctgctatgt ggcgcggtat tatcccgtat tgacgccggg
     5401 caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga gtactcacca
     5461 gtcacagaaa agcatcttac ggatggcatg acagtaagag aattatgcag tgctgccata
     5521 accatgagtg ataacactgc ggccaactta cttctgacaa cgatcggagg accgaaggag
     5581 ctaaccgctt ttttgcacaa catgggggat catgtaactc gccttgatcg ttgggaaccg
     5641 gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgt agcaatggca
     5701 acaacgttgc gcaaactatt aactggcgaa ctacttactc tagcttcccg gcaacaatta
     5761 atagactgga tggaggcgga taaagttgca ggaccacttc tgcgctcggc ccttccggct
     5821 ggctggttta ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca
     5881 gcactggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac ggggagtcag
     5941 gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact gattaagcat
     6001 tggtaactgt cagaccaagt ttactcatat atactttaga ttgatttaaa acttcatttt
     6061 taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa
     6121 cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga
     6181 gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg
     6241 gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc
     6301 agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca ccacttcaag
     6361 aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc
     6421 agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg
     6481 cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac
     6541 accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga
     6601 aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt
     6661 ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag
     6721 cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg
     6781 gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta
     6841 tcccctgatt ctgtggataa ccgtattacc gcctttgagt gagctgatac cgctcgccgc
     6901 agccgaacga ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg cccaatacgc
     6961 aaaccgcctc tccccgcgcg ttggccgatt cattaatgca gctggcacga caggtttccc
     7021 gactggaaag cgggcagtga gcgcaacgca attaatgtga gttagctcac tcattaggca
     7081 ccccaggctt tacactttat gcttccggct cgtatgttgt gtggaattgt gagcggataa
     7141 caatttcaca caggaaacag ctatgaccat gattacgcca agcgcgcaat taaccctcac
     7201 taaagggaac aaaagctgga gctccggatt agaggccgcc gagcgggtga cagccctccg
     7261 aaggaagact ctcctccgtg cgtcctcgtc ttcaccggtc gcgttcctga aacgcagatg
     7321 tgcctcgcgc cgcactgctc cgaacaataa agattctaca atactagctt ttatggttat
     7381 gaagaggaaa aattggcagt aacctggccc cacaaacctt caaatgaacg aatcaaatta
     7441 acaaccatag gatgataatg cgattagttt tttagcctta tttctggggt aattaatcag
     7501 cgaagcgatg atttttgatc tattaacaga tatataaatg caaaaactgc ataaccactt
     7561 taactaatac tttcaacatt ttcggtttgt attacttctt attcaaatgt aataaaagta
     7621 tcaacaaaaa attgttaata tacctctata ctttaacgtc aaggag
//
