LOCUS       sequence-417765-        9801 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Yeast integrative vector for expression of Nicastrin and APH-1a-HA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9801)
  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 9801)
  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: pRS402-nicastrin/APH-1a-HA
FEATURES             Location/Qualifiers
     source          1..9801
                     /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"
     CDS             complement(2285..2311)
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     promoter        complement(3138..3481)
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     promoter        3488..4154
                     /gene="S. cerevisiae TDH3"
                     /label=GAP promoter
                     /note="promoter for glyceraldehyde-3-phosphate 
                     dehydrogenase; also known as the TDH3 promoter"
     primer_bind     4024..4047
                     /label=GPDpro-F
                     /note="S. cerevisiae GPD promoter, forward primer"
     primer_bind     complement(6516..6540)
                     /label=pHybLex-R
                     /note="ADH terminator, reverse primer"
     rep_origin      6690..7145
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6777..6796)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6987..7008
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(7350..9065)
                     /codon_start=1
                     /gene="S. cerevisiae ADE2"
                     /product="phosphoribosylaminoimidazole carboxylase, 
                     required for adenine biosynthesis"
                     /label=ADE2
                     /note="yeast auxotrophic marker"
                     /translation="MDSRTVGILGGGQLGRMIVEAANRLNIKTVILDAENSPAKQISNS
                     NDHVNGSFSNPLDIEKLAEKCDVLTIEIEHVDVPTLKNLQVKHPKLKIYPSPETIRLIQ
                     DKYIQKEHLIKNGIAVTQSVPVEQASETSLLNVGRDLGFPFVLKSRTLAYDGRGNFVVK
                     NKEMIPEALEVLKDRPLYAEKWAPFTKELAVMIVRSVNGLVFSYPIVETIHKDNICDLC
                     YAPARVPDSVQLKAKLLAENAIKSFPGCGIFGVEMFYLETGELLINEIAPRPHNSGHYT
                     IDACVTSQFEAHLRSILDLPMPKNFTSFSTITTNAIMLNVLGDKHTKDKELETCERALA
                     TPGSSVYLYGKESRPNRKVGHINIIASSMAECEQRLNYITGRTDIPIKISVAQKLDLEA
                     MVKPLVGIIMGSDSDLPVMSAACAVLKDFGVPFEVTIVSAHRTPHRMSAYAISASKRGI
                     KTIIAGAGGAAHLPGMVAAMTPLPVIGVPVKGSCLDGVDSLHSIVQMPRGVPVATVAIN
                     NSTNAALLAVRLLGAYDSSYTTKMEQFLLKQEEEVLVKAQKLETVGYEAYLENK"
     promoter        complement(9066..9441)
                     /gene="S. cerevisiae ADE2"
                     /label=ADE2 promoter
     primer_bind     complement(9632..9651)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     9751..9773
                     /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 gttaggcata gtcagggacg tcataaggat aagaagtatc ttcaggtggt atacgaagag
     2341 ctgagtaaac catgacacga gaatcttctt gacgtctaca tagtagagaa cgttggatag
     2401 accttaggct accgcctgca gtaataaacg cccaaagacc catactgact gttacagcgt
     2461 atataggtaa taaactagct tcgtaccatg gattaagaaa agtaagaccg gaagttaaca
     2521 gatggctacc gacgaccaaa cccagagccc aatatctacg tctttcgcat gcgtcgaaga
     2581 atactacgcc ccagaaagtg tgaagtaaaa ttatagcggc agtcaagaaa gcactggtta
     2641 agaagtagta cggcgaatca ccgtgtattc cgactacgcc aggacccaac gcgtctgcca
     2701 gtatgtttat tacgctaaac accccagaaa ttattccaaa tgatagtccg cttacgtacg
     2761 ccatttgtct tatactaatc ggcgacctac catcttcgga taagctggcc aagccttcgt
     2821 cagctttctt cagtagttta taataagcga atctaaatac ttcttgtaac aatacgctta
     2881 ccgctgcgcc gaagattaat aacccatatt gtagtctcgc gtcacttcta tcagttacgt
     2941 gaacaagaat aaaccaaaca actgacgcca ataacaaaga tactaaccaa aagaaggcac
     3001 cagcaactag aattatgact cttaatggat cacctgctac tgttattaga aacaatgcaa
     3061 atgcgggtcc aaatgcaaca aaggtacatc caaagaaaac tgcggcaccc attttttttg
     3121 tcgacgggcc cggatccggt tgtttatgtt cggatgtgat gtgagaactg tatcctagca
     3181 agattttaaa aggaagtata tgaaagaaga acctcagtgg caaatcctaa ccttttatat
     3241 ttctctacag gggcgcggcg tggggacaat tcaacgcgtc tgtgagggga gcgtttccct
     3301 gctcgcaggt ctgcagcgag gagccgtaat ttttgcttcg cgccgtgcgg ccatcaaaat
     3361 gtatggatgc aaatgattat acatggggat gtatgggcta aatgtacggg cgacagtcac
     3421 atcatgcccc tgagctgcgc acgtcaagac tgtcaaggag ggtattctgg gcctccatgt
     3481 caccggttca ttatcaatac tgccatttca aagaatacgt aaataattaa tagtagtgat
     3541 tttcctaact ttatttagtc aaaaaattag ccttttaatt ctgctgtaac ccgtacatgc
     3601 ccaaaatagg gggcgggtta cacagaatat ataacatcgt aggtgtctgg gtgaacagtt
     3661 tattcctggc atccactaaa tataatggag cccgcttttt aagctggcat ccagaaaaaa
     3721 aaagaatccc agcaccaaaa tattgttttc ttcaccaacc atcagttcat aggtccattc
     3781 tcttagcgca actacagaga acaggggcac aaacaggcaa aaaacgggca caacctcaat
     3841 ggagtgatgc aacctgcctg gagtaaatga tgacacaagg caattgaccc acgcatgtat
     3901 ctatctcatt ttcttacacc ttctattacc ttctgctctc tctgatttgg aaaaagctga
     3961 aaaaaaaggt tgaaaccagt tccctgaaat tattccccta cttgactaat aagtatataa
     4021 agacggtagg tattgattgt aattctgtaa atctatttct taaacttctt aaattctact
     4081 tttatagtta gtcttttttt tagttttaaa acaccaagaa cttagtttcg aataaacaca
     4141 cataaacaaa caaagaattc agcggccgca ctagtatcga tatggcaact gcgggaggag
     4201 gaagtggcgc cgatccagga tctagagggt tattaaggtt attatccttt tgtgtattgt
     4261 tggcggggtt atgtagaggc aatagcgtcg aacgtaaaat ctacattcct ctaaacaaga
     4321 ccgcaccttg cgtaagattg ttgaatgcta cgcaccaaat aggttgtcaa tcaagcatct
     4381 ctggtgatac cggtgtgata catgttgttg aaaaggaaga agatttacaa tgggttttaa
     4441 cggacggtcc aaatccaccg tatatggtct tactagaatc caaacacttc actagagacc
     4501 taatggaaaa gctaaagggt aggacatcta gaatagcagg cttggccgtc agcctgacta
     4561 aaccttcacc agcatctgga ttttccccca gcgtgcaatg tcctaacgac ggcttcggag
     4621 tatattcgaa ctcatacgga ccggaatttg cacattgtcg tgagatccaa tggaactctt
     4681 tgggtaacgg cttagcgtac gaggatttct cgtttccaat tttcctgttg gaagacgaga
     4741 acgagacgaa ggtaattaaa caatgttacc aggaccataa tctatctcaa aacgggagtg
     4801 cccccacttt tccgttgtgc gcaatgcaat tattctcgca catgcacgca gtaattagta
     4861 ccgcaacatg tatgagaagg tcatctattc agtcaacgtt ttctataaat cccgagatag
     4921 tgtgcgatcc tttgagtgac tataacgtat ggtccatgtt aaaacccatc aacactacag
     4981 gtacactaaa accagatgat agagtagttg tagccgcaac tagactagac tctagaagtt
     5041 tcttttggaa cgttgcgcct ggtgcggaga gtgctgtcgc atctttcgtt acgcaacttg
     5101 ccgcagccga ggcccttcag aaagctccgg acgtaacgac gttgcctagg aacgtaatgt
     5161 tcgtattctt ccagggtgag accttcgatt atataggttc gagcagaatg gtttatgaca
     5221 tggagaaagg aaaattccca gttcaactag aaaacgtaga ttcgttcgta gaactaggtc
     5281 aagttgcact taggacctct ctggaattat ggatgcatac cgacccggta agtcaaaaga
     5341 acgaaagtgt gagaaatcaa gttgaagact tgttggcaac cttagaaaag tctggcgccg
     5401 gggttccagc cgttatatta agaagaccta accaatcgca accattaccc ccttcatctt
     5461 tgcaaagatt cttaagagcc agaaatattt cgggtgtggt cttagcggat cattcagggg
     5521 cttttcacaa taagtactat caatctatct atgatacggc ggaaaatatc aacgtttcct
     5581 acccggagtg gttaagtcca gaggaagatt tgaatttcgt tactgataca gctaaagcat
     5641 tggcggacgt tgcaactgta cttggtagag ccctttacga attagctggt ggtacaaatt
     5701 tctctgatac cgtacaagca gacccacaga cagtgactcg tttactatac ggatttctga
     5761 tcaaggctaa taattcgtgg tttcaatcta ttttaagaca agatttgagg tcttatctgg
     5821 gagatggccc attgcagcac tatattgcag tgagttctcc aactaataca acgtacgtag
     5881 ttcaatacgc tctggccaac ctaacgggaa ccgtcgttaa tttaacaaga gaacaatgtc
     5941 aagacccttc aaaggttcct tcggagaata aagacttata cgaatattcg tgggtgcaag
     6001 gtccattaca cagtaacgaa actgatcgtt taccaaggtg cgtcagatcc accgccaggc
     6061 ttgctcgtgc tttatcccca gctttcgagt taagccaatg gtcaagcaca gagtatagta
     6121 cttggaccga atcgaggtgg aaggacatta gagccaggat tttcttaatt gcatcaaagg
     6181 agctagaact aataactttg accgtgggat ttggtatttt aattttcagc ttaattgtca
     6241 catactgtat aaacgctaag gcagacgttt tatttatcgc accacgtgaa ccgggcgcag
     6301 tctcctatta aagatctgag ctcttaatta acaattcttc gccagaggtt tggtcaagtc
     6361 tccaatcaag gttgtcggct tgtctacctt gccagaaatt tacgaaaaga tggaaaaggg
     6421 tcaaatcgtt ggtagatacg ttgttgacac ttctaaataa gcgaatttct tatgatttat
     6481 gatttttatt attaaataag ttataaaaaa aataagtgta tacaaatttt aaagtgactc
     6541 ttaggtttta aaacgaaaat tcttattctt gagtaactct ttcctgtagg tcaggttgct
     6601 ttctcaggta tagcaattcc tggcgtaata gcgaagaggc ccgcaccgat cgcccttccc
     6661 aacagttgcg cagcctgaat ggcgaatgga cgcgccctgt agcggcgcat taagcgcggc
     6721 gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc
     6781 tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa
     6841 tcgggggctc cctttagggt tccgatttag tgctttacgg cacctcgacc ccaaaaaact
     6901 tgattagggt gatggttcac gtagtgggcc atcgccctga tagacggttt ttcgcccttt
     6961 gacgttggag tccacgttct ttaatagtgg actcttgttc caaactggaa caacactcaa
     7021 ccctatctcg gtctattctt ttgatttata agggattttg ccgatttcgg cctattggtt
     7081 aaaaaatgag ctgatttaac aaaaatttaa cgcgaatttt aacaaaatat taacgcttac
     7141 aatttcctga tgcggtattt tctccttacg catctgtgcg gtatttcaca ccgcatagat
     7201 cttatgtatg aaattcttaa aaaaggacac ctgtaagcgt tgatttctat gtatgaagtc
     7261 cacatttgat gtaatcataa caaagcctaa aaaataggta tatcatttta taattatttg
     7321 ctgtacaagt atatcaataa acttatatat tacttgtttt ctagataagc ttcgtaaccg
     7381 acagtttcta acttttgtgc tttgacaaga acttcttctt cttgctttaa taaaaactgt
     7441 tccattttcg ttgtataact tgaatcataa gcgccaagca gtctgacagc caacagcgca
     7501 gcgttcgtac tattattaat agcgacggta gctactggaa cacctctagg catttgcaca
     7561 attgaatgta aagaatctac tccatctaga caagaacctt ttacgggcac accgatgaca
     7621 ggaagtggtg tcattgcagc caccatacct ggcaagtgag cagccccacc agctccagcg
     7681 ataattgttt taattccacg cttgcttgcg gaaatagcat atgctgacat cctatgtgga
     7741 gttctatgag cagagactat tgtcacttca aatggaacgc caaaatcttt taaaaccgca
     7801 catgcggcag acattaccgg caagtcagag tctgatccca tgatgattcc aaccaatggt
     7861 ttgaccattg cttccaagtc caacttttga gcgacagaga ttttgattgg aatatcagtt
     7921 ctacctgtaa tgtagttcag cctttgttca cattccgcca tactggaggc aataatattt
     7981 atgtgaccta cttttctgtt aggtctagac tcttttccat ataagtacac tgaggaacct
     8041 ggagtcgcca atgctctttc gcaagtttct agctctttat cttttgtatg tttgtctcca
     8101 agaacattta gcataatggc gttcgttgta atggtggaga aagatgtgaa attctttggc
     8161 attggcaaat ccaatattga tctcaaatga gcttcaaatt gagaagtgac gcaagcatca
     8221 atggtataat gtccagagtt gtgaggcctt ggggcaattt cgttaataag caattcccct
     8281 gtttctaaat agaacatttc cacaccaaat ataccacaac cgggaaaaga tttgattgca
     8341 ttttctgcca acaacttcgc cttaagttga acggagtccg gaactctagc aggcgcataa
     8401 cataagtcac aaatattgtc cttgtggata gtctctacaa ttgggtaaga aaacactaaa
     8461 ccgttaacag acctcacaat catgactgct aattctttag taaatggtgc ccatttttcg
     8521 gcgtacaaag gacgatcctt cagtacttcc aaagcttccg gaatcatttc cttattcttt
     8581 acaacgaagt tacctcttcc atcgtatgcc aaagtcctcg acttcaagac gaatggaaaa
     8641 cccaaatctc ttccaacatt caatagggac gtctcactgg cttgttccac aggaacactt
     8701 tgggtaactg ctataccatt tttgattaaa tgctcttttt gaatatattt gtcttgtatc
     8761 aatctgattg tttctggaga agggtaaatt tttaatttgg gatgttttac ttgaagattc
     8821 tttagtgtag gaacatcaac atgctcaatc tcaatcgtta gcacatcaca tttttcagct
     8881 agtttttcga tatcaagagg attggaaaag gagccattaa cgtggtcatt ggagttgctt
     8941 atttgtttgg caggagaatt ttcagcatct agtattaccg tcttaatgtt gagcctgttt
     9001 gctgcctcaa caatcatacg tcccaattgt ccccctccta atataccaac tgttctagaa
     9061 tccatacttg attgttttgt ccgattttct tgtttttctt gattgttata gtaggatgta
     9121 cttagaagag agatccaacg attttacgca ccaatttata catgaaatgc tccataatat
     9181 tgtccattta gttcttaata aaaggtcagc aagagtcaat cacttagtat tacccggttc
     9241 gtagccatgc aacaagagtc atttgtcagc atagctgtaa taatcaatca tgacgtaaga
     9301 aatgtatcat aattaaaagt tgttaaagat gtcagtgtta tgttggtgtt acaaaattct
     9361 cggcttctca ctaatattta atatctctta aattttatct gtctttgatt cttttaagaa
     9421 aagttatgta ttattcaaga attaattcag atctattctt ttgatttata agggattttg
     9481 ccgatttcgg cctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaatttt
     9541 aacaaaatat taacgcttac aatttcctga tgcggtattt tctccttacg catctgtgcg
     9601 gtatttcaca ccgcatatgg tgcactctca gtacaatctg ctctgatgcc gcatagttaa
     9661 gccagccccg acacccgcca acacccgctg acgcgccctg acgggcttgt ctgctcccgg
     9721 catccgctta cagacaagct gtgaccgtct ccgggagctg catgtgtcag aggttttcac
     9781 cgtcatcacc gaaacgcgcg a
//
