LOCUS       sequence-417771-        8404 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Yeast integrative vector for expression of FlagPEN2 and Presenilin 
            1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8404)
  AUTHORS   Ortner M, Guschtschin-Schmidt N, Stelzer W, Muhle-Goll C, Langosch D
  TITLE     Permissive Conformations of a Transmembrane Helix Allow 
            Intramembrane Proteolysis by gamma-Secretase.
  JOURNAL   J Mol Biol. 2023 Aug 1;435(18):168218. doi: 
            10.1016/j.jmb.2023.168218.
  PUBMED    37536392
REFERENCE   2  (bases 1 to 8404)
  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: "J Mol 
            Biol."; date: "2023-08-1"; volume: "435(18):168218. doi"; pages: " 
            10.1016/j.jmb.2023.168218"
COMMENT     Sequence Label: pRS405-FlagPEN2/PS1wt
FEATURES             Location/Qualifiers
     source          1..8404
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(10..28)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        96..200
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(201..269,270..1061)
                     /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(419..438)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1232..1820
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1721..1740
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1974..1991
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(2060..2249)
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(3687..3706)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3688..3706)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        complement(3714..4057)
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     promoter        4064..4730
                     /gene="S. cerevisiae TDH3"
                     /label=GAP promoter
                     /note="promoter for glyceraldehyde-3-phosphate 
                     dehydrogenase; also known as the TDH3 promoter"
     primer_bind     4600..4623
                     /label=GPDpro-F
                     /note="S. cerevisiae GPD promoter, forward primer"
     CDS             4775..4798
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     primer_bind     complement(5309..5333)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     rep_origin      5483..5938
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5570..5589)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5780..5801
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        6239..6646
                     /gene="S. cerevisiae LEU2"
                     /label=LEU2 promoter
     CDS             6647..7741
                     /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(8235..8254)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     8354..8376
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
ORIGIN
        1 gacgaaaggg cctcgtgata cgcctatttt tataggttaa tgtcatgata ataatggttt
       61 cttagacgtc aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt
      121 tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat
      181 aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt
      241 ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg
      301 ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga
      361 tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc
      421 tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga gcaactcggt cgccgcatac
      481 actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg
      541 gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca
      601 acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg
      661 gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg
      721 acgagcgtga caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg
      781 gcgaactact tactctagct tcccggcaac aattaataga ctggatggag gcggataaag
      841 ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg
      901 gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct
      961 cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac
     1021 agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac caagtttact
     1081 catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga
     1141 tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt
     1201 cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct
     1261 gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc
     1321 taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc
     1381 ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc
     1441 tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg
     1501 ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt
     1561 cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg
     1621 agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg
     1681 gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt
     1741 atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag
     1801 gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt
     1861 gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta
     1921 ttaccgcctt tgagtgagct gataccgctc gccgcagccg aacgaccgag cgcagcgagt
     1981 cagtgagcga ggaagcggaa gagcgcccaa tacgcaaacc gcctctcccc gcgcgttggc
     2041 cgattcatta atgcagcgac ttcgagcgtc ccaaaacctt ctcaagcaag gttttcagta
     2101 taatgttaca tgcgtacacg cgtctgtaca gaaaaaaaag aaaaatttga aatataaata
     2161 acgttcttaa tactaacata actataaaaa aataaatagg gacctagact tcaggttgtc
     2221 taactccttc cttttcggtt agagcggatc ttagctagcc gcggtaccaa gcttactcga
     2281 gtcaaatgta gaactggtga aaggccagct gatccatgaa tggttggact aggtagtcgg
     2341 tcgcgaaata aaataccagt ccgaaagtta tcgaaatagg caatgcaggt aacgccttct
     2401 taaatatggc taacagcaat agagttaaac ataacccgat caaaattgca acaaagcaag
     2461 caatagtagt attccaatca ccggatgccg tagcactcgc cttcccgaca agcactgaat
     2521 aaaagataaa gtcacctaga cctaacttaa cacctctctc ttccggatct tctccggcta
     2581 atattgaact actcagctct tgtacggctg ccctactttc tggggttgat ctgtgcggac
     2641 caaggtgaga atctctttga gcttcccact cttcagaaaa tccaccgtcg tcgttttccg
     2701 ctaccgtatc ctgagattct ctctcggtac tctcagcgtt gtattttgag ttcttagaaa
     2761 cccttctctg tgcctcagga tcaccctcag ccatgttaac tagccatacc atggtcgaag
     2821 aatatatcaa ggcagggaat aaagtctcgt tacgttcttg ggcggtctcc accaacattc
     2881 ttaacggccc ctttgggcat agcactgcga ctaggtcgta gacgcttata actgccagta
     2941 tcaaccatgc cgtccactct ggcaaatatt ttatgaaaac taaagccatt aaagctgaga
     3001 tcatgattaa gtacgcttgt tgcagtctta acgggccttt ccaatggatc gagatcatac
     3061 cgactactcc aaggttccaa atcaacaaag ctacggttat ataatcaacg gcgacattgt
     3121 atgtcttgaa gacctcaccc agatagataa aggagaagaa gaataacagc agtaagctag
     3181 agatgattag ccatgcgtga ataaccttgt aacatctata cttgtaaagg acgacaagta
     3241 aaatcgtcat caccacgatt acgctaatca taatcgccgc gttcaatata ctatgtaacg
     3301 cacgttgacc tactgtttcc gtgtcctcag taaacggagt gtatatcaat tgaccgtctt
     3361 ttctagtgta gaaactaaca cttttgatag ttgcaactac gactaccata cacaaggtga
     3421 ctggaacgaa caacataatt acgtgtttcg ccccgtactt taaagttaat tcttcgtctt
     3481 cctcttcgtc ctgttctaca acttgacgtg aattgccttg gggcctgccg ttactaagag
     3541 gttctggatg tcctaaagaa cgcctatcat tatgttcttg tctctccctg ttatcgttgg
     3601 tgtttgataa atgattatct tcggacattt gcgcgttttg aaaataactt aagggggctg
     3661 gaagttcagt catgtcgacg cccgggccct atagtgagtc gtattacgga tccggttgtt
     3721 tatgttcgga tgtgatgtga gaactgtatc ctagcaagat tttaaaagga agtatatgaa
     3781 agaagaacct cagtggcaaa tcctaacctt ttatatttct ctacaggggc gcggcgtggg
     3841 gacaattcaa cgcgtctgtg aggggagcgt ttccctgctc gcaggtctgc agcgaggagc
     3901 cgtaattttt gcttcgcgcc gtgcggccat caaaatgtat ggatgcaaat gattatacat
     3961 ggggatgtat gggctaaatg tacgggcgac agtcacatca tgcccctgag ctgcgcacgt
     4021 caagactgtc aaggagggta ttctgggcct ccatgtcacc ggttcattat caatactgcc
     4081 atttcaaaga atacgtaaat aattaatagt agtgattttc ctaactttat ttagtcaaaa
     4141 aattagcctt ttaattctgc tgtaacccgt acatgcccaa aatagggggc gggttacaca
     4201 gaatatataa catcgtaggt gtctgggtga acagtttatt cctggcatcc actaaatata
     4261 atggagcccg ctttttaagc tggcatccag aaaaaaaaag aatcccagca ccaaaatatt
     4321 gttttcttca ccaaccatca gttcataggt ccattctctt agcgcaacta cagagaacag
     4381 gggcacaaac aggcaaaaaa cgggcacaac ctcaatggag tgatgcaacc tgcctggagt
     4441 aaatgatgac acaaggcaat tgacccacgc atgtatctat ctcattttct tacaccttct
     4501 attaccttct gctctctctg atttggaaaa agctgaaaaa aaaggttgaa accagttccc
     4561 tgaaattatt cccctacttg actaataagt atataaagac ggtaggtatt gattgtaatt
     4621 ctgtaaatct atttcttaaa cttcttaaat tctactttta tagttagtct tttttttagt
     4681 tttaaaacac caagaactta gtttcgaata aacacacata aacaaacaaa gaattcaacc
     4741 ctcactaaag ggcggccgca ctagtatcga tatggattat aaggatgacg atgataaaat
     4801 gaatctagaa agggttagca acgaggaaaa gctaaacttg tgtcgtaagt attacttggg
     4861 tggtttcgca ttccttccat tcttatggtt agtaaatata ttctggttct ttagagaggc
     4921 ttttctagtt cctgcatata ctgagcaaag tcagattaag ggttacgttt ggagatccgc
     4981 agtcggtttc ttattttggg ttatcgtcct tacttcatgg ataacaatat ttcaaattta
     5041 tcgtcctaga tggggcgctc taggggatta tttaagtttt acgattccac taggtacacc
     5101 ttaaagatct gagctcttaa ttaacaattc ttcgccagag gtttggtcaa gtctccaatc
     5161 aaggttgtcg gcttgtctac cttgccagaa atttacgaaa agatggaaaa gggtcaaatc
     5221 gttggtagat acgttgttga cacttctaaa taagcgaatt tcttatgatt tatgattttt
     5281 attattaaat aagttataaa aaaaataagt gtatacaaat tttaaagtga ctcttaggtt
     5341 ttaaaacgaa aattcttatt cttgagtaac tctttcctgt aggtcaggtt gctttctcag
     5401 gtatagcaat tcctggcgta atagcgaaga ggcccgcacc gatcgccctt cccaacagtt
     5461 gcgcagcctg aatggcgaat ggacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg
     5521 gtggttacgc gcagcgtgac cgctacactt gccagcgccc tagcgcccgc tcctttcgct
     5581 ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggg
     5641 ctccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgattag
     5701 ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc tttgacgttg
     5761 gagtccacgt tctttaatag tggactcttg ttccaaactg gaacaacact caaccctatc
     5821 tcggtctatt cttttgattt ataagggatt ttgccgattt cggcctattg gttaaaaaat
     5881 gagctgattt aacaaaaatt taacgcgaat tttaacaaaa tattaacgct tacaatttcc
     5941 tgatgcggta ttttctcctt acgcatctgt gcggtatttc acaccgcata tcgaccggtc
     6001 gaggagaact tctagtatat ctacatacct aatattattg ccttattaaa aatggaatcc
     6061 caacaattac atcaaaatcc acattctctt caaaatcaat tgtcctgtac ttccttgttc
     6121 atgtgtgttc aaaaacgtta tatttatagg ataattatac tctatttctc aacaagtaat
     6181 tggttgtttg gccgagcggt ctaaggcgcc tgattcaaga aatatcttga ccgcagttaa
     6241 ctgtgggaat actcaggtat cgtaagatgc aagagttcga atctcttagc aaccattatt
     6301 tttttcctca acataacgag aacacacagg ggcgctatcg cacagaatca aattcgatga
     6361 ctggaaattt tttgttaatt tcagaggtcg cctgacgcat ataccttttt caactgaaaa
     6421 attgggagaa aaaggaaagg tgagagcgcc ggaaccggct tttcatatag aatagagaag
     6481 cgttcatgac taaatgcttg catcacaata cttgaagttg acaatattat ttaaggacct
     6541 attgtttttt ccaataggtg gttagcaatc gtcttacttt ctaacttttc ttacctttta
     6601 catttcagca atatatatat atatatttca aggatatacc attctaatgt ctgcccctaa
     6661 gaagatcgtc gttttgccag gtgaccacgt tggtcaagaa atcacagccg aagccattaa
     6721 ggttcttaaa gctatttctg atgttcgttc caatgtcaag ttcgatttcg aaaatcattt
     6781 aattggtggt gctgctatcg atgctacagg tgttccactt ccagatgagg cgctggaagc
     6841 ctccaagaag gctgatgccg ttttgttagg tgctgtgggt ggtcctaaat ggggtaccgg
     6901 tagtgttaga cctgaacaag gtttactaaa aatccgtaaa gaacttcaat tgtacgccaa
     6961 cttaagacca tgtaactttg catccgactc tcttttagac ttatctccaa tcaagccaca
     7021 atttgctaaa ggtactgact tcgttgttgt cagagaatta gtgggaggta tttactttgg
     7081 taagagaaag gaagacgatg gtgatggtgt cgcttgggat agtgaacaat acaccgttcc
     7141 agaagtgcaa agaatcacaa gaatggccgc tttcatggcc ctacaacatg agccaccatt
     7201 gcctatttgg tccttggata aagctaatgt tttggcctct tcaagattat ggagaaaaac
     7261 tgtggaggaa accatcaaga acgaattccc tacattgaag gttcaacatc aattgattga
     7321 ttctgccgcc atgatcctag ttaagaaccc aacccaccta aatggtatta taatcaccag
     7381 caacatgttt ggtgatatca tctccgatga agcctccgtt atcccaggtt ccttgggttt
     7441 gttgccatct gcgtccttgg cctctttgcc agacaagaac accgcatttg gtttgtacga
     7501 accatgccac ggttctgctc cagatttgcc aaagaataag gtcaacccta tcgccactat
     7561 cttgtctgct gcaatgatgt tgaaattgtc attgaacttg cctgaagaag gtaaggccat
     7621 tgaagatgca gttaaaaagg ttttggatgc aggtatcaga actggtgatt taggtggttc
     7681 caacagtacc accgaagtcg gtgatgctgt cgccgaagaa gttaagaaaa tccttgctta
     7741 aaaagattct ctttttttat gatatttgta cataaacttt ataaatgaaa ttcataatag
     7801 aaacgacacg aaattacaaa atggaatatg ttcatagggt agacgaaact atatacgcaa
     7861 tctacataca tttatcaaga aggagaaaaa ggaggatgta aaggaataca ggtaagcaaa
     7921 ttgatactaa tggctcaacg tgataaggaa aaagaattgc actttaacat taatattgac
     7981 aaggaggagg gcaccacaca aaaagttagg tgtaacagaa aatcatgaaa ctatgattcc
     8041 taatttatat attggaggat tttctctaaa aaaaaaaaaa tacaacaaat aaaaaacact
     8101 caatgacctg accatttgat ggagtttaag tcaatacctt cttgaaccat ttcccataat
     8161 ggtgaaagtt ccctcaagaa ttttactctg tcagaaacgg ccttaacgac gtagtcgata
     8221 tggtgcactc tcagtacaat ctgctctgat gccgcatagt taagccagcc ccgacacccg
     8281 ccaacacccg ctgacgcgcc ctgacgggct tgtctgctcc cggcatccgc ttacagacaa
     8341 gctgtgaccg tctccgggag ctgcatgtgt cagaggtttt caccgtcatc accgaaacgc
     8401 gcga
//
