LOCUS       sequence-460494-        6268 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Yeast optimized human Amyloid beta-42 fused with aga2 protein gene 
            under gal1 promoter for the expression of Amyloid beta-42 in yeast 
            surface display..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6268)
  AUTHORS   Ozcelik CE, Begli O, Hincer A, Ahan RE, Kesici MS, Oguz O, Kasirga 
            TS, Ozcubukcu S, Seker UOS
  TITLE     Synergistic Screening of Peptide-Based Biotechnological Drug 
            Candidates for Neurodegenerative Diseases Using Yeast Display and 
            Phage Display.
  JOURNAL   ACS Chem Neurosci. 2023 Oct 4;14(19):3609-3621. doi: 
            10.1021/acschemneuro.3c00248. Epub 2023 Aug 28.
  PUBMED    37638647
REFERENCE   2  (bases 1 to 6268)
  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: 
            "10.1021/acschemneuro.3c00248"; journalName: "ACS Chem Neurosci"; 
            date: "2023-10-4- 4"; volume: "14"; issue: "19"; pages: "3609-3621"
COMMENT     Sequence Label: pETcon-pGAL1-Aga2-Amyloid beta-42-c_myc
FEATURES             Location/Qualifiers
     source          1..6268
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    2..18
                     /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     7..29
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     26..42
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     26..42
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        63..81
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        111..763
                     /gene="GAL1 and GAL10 genes of S. cerevisiae"
                     /label=GAL1,10 promoter
                     /note="divergent inducible promoter, regulated by Gal4"
     protein_bind    315..432
                     /label=UAS
                     /bound_moiety="Gal4"
                     /note="upstream activating sequence mediating 
                     Gal4-dependent induction"
     primer_bind     727..750
                     /label=GAL1
                     /note="S. cerevisiae GAL1 promoter, forward primer"
     CDS             1082..1093
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     CDS             1094..1120
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             1331..1360
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     primer_bind     complement(1689..1708)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1690..1708)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1715..1732)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1715..1731)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1724..1746)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      1872..2327
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1959..1978)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2169..2190
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(2430..3104)
                     /codon_start=1
                     /gene="S. cerevisiae TRP1"
                     /product="phosphoribosylanthranilate isomerase, required 
                     for tryptophan biosynthesis"
                     /label=TRP1
                     /note="yeast auxotrophic marker"
                     /translation="MSVINFTGSSGPLVKVCGLQSTEAAECALDSDADLLGIICVPNRK
                     RTIDPVIARKISSLVKAYKNSSGTPKYLVGVFRNQPKEDVLALVNDYGIDIVQLHGDES
                     WQEYQEFLGLPVIKRLVFPKDCNILLSAASQKPHSFIPLFDSEAGGTGELLDWNSISDW
                     VGRQESPESLHFMLAGGLTPENVGDALRLNGVIGVDVSGGVETNGVKDSNKIANFVKNA
                     KK"
     promoter        complement(3105..3385)
                     /gene="S. cerevisiae TRP1"
                     /label=TRP1 promoter
     primer_bind     complement(3406..3425)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3525..3547
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3585..3603)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     misc_feature    3644..4147
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     promoter        4184..4288
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4289..4357,4358..5149)
                     /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(4507..4526)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5320..5908
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5809..5828
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6062..6079
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    6196..6217
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6232..6237,6238..6255,6256..6262)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
ORIGIN
        1 attgtgagcg gataacaatt tcacacagga aacagctatg accatgatta cgccaagctc
       61 gaaattaacc ctcactaaag ggaacaaaag ctggtaccaa ttccttgaat tttcaaaaat
      121 tcttactttt tttttggatg gacgcaaaga agtttaataa tcatattaca tggcattacc
      181 accatataca tatccatatc taatcttact tatatgttgt ggaaatgtaa agagccccat
      241 tatcttagcc taaaaaaacc ttctctttgg aactttcagt aatacgctta actgctcatt
      301 gctatattga agtacggatt agaagccgcc gagcgggtga cagccctccg aaggaagact
      361 ctcctccgtg cgtcctcgtc ttcaccggtc gcgttcctga aacgcagatg tgcctcgcgc
      421 cgcactgctc cgaacaataa agattctaca atactagctt ttatggttat gaagaggaaa
      481 aattggcagt aacctggccc cacaaacctt caaatgaacg aatcaaatta acaaccatag
      541 gatgataatg cgattagttt tttagcctta tttctggggt aattaatcag cgaagcgatg
      601 atttttgatc tattaacaga tatataaatg caaaaactgc ataaccactt taactaatac
      661 tttcaacatt ttcggtttgt attacttctt attcaaatgt aataaaagta tcaacaaaaa
      721 attgttaata tacctctata ctttaacgtc aaggagaaaa aaccccggat cgaattccct
      781 acttcataca ttttcaatta agatgcagtt acttcgctgt ttttcaatat tttctgttat
      841 tgcttcagtt ttagcacagg aactgacaac tatatgcgag caaatcccct caccaacttt
      901 agaatcgacg ccgtactctt tgtcaacgac tactattttg gccaacggga aggcaatgca
      961 aggagttttt gaatattaca aatcagtaac gtttgtcagt aattgcggtt ctcacccctc
     1021 aacaactagc aaaggcagcc ccataaacac acagtatgtt tttaaggaca atagctcgac
     1081 gattgaaggt agatacccat acgacgttcc agactacgct ctgcaggcta gtggtggagg
     1141 aggctctggt ggaggcggta gcggaggcgg agggtcgact agtatggacg cggagtttcg
     1201 tcacgatagt ggatatgagg ttcatcatca aaagttagtc ttctttgcgg aggatgtagg
     1261 ttctaataaa ggtgccataa tcggactaat ggtcggcgga gttgtgatag ctctcgaggg
     1321 gggcggatcc gaacaaaagc ttatttctga agaggacttg taatagagat ctgataacaa
     1381 cagtgtagat gtaacaaaat cgactttgtt cccactgtac ttttagctcg tacaaaatac
     1441 aatatacttt tcatttctcc gtaaacaaca tgttttccca tgtaatatcc ttttctattt
     1501 ttcgttccgt taccaacttt acacatactt tatatagcta ttcacttcta tacactaaaa
     1561 aactaagaca attttaattt tgctgcctgc catatttcaa tttgttataa attcctataa
     1621 tttatcctat tagtagctaa aaaaagatga atgtgaatcg aatcctaaga gaattgagct
     1681 ccaattcgcc ctatagtgag tcgtattaca attcactggc cgtcgtttta caacgtcgtg
     1741 actgggaaaa ccctggcgtt acccaactta atcgccttgc agcacatccc cctttcgcca
     1801 gctggcgtaa tagcgaagag gcccgcaccg atcgcctttc ccaacagttg cgcagcctga
     1861 atggcgaatg gacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg
     1921 cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc
     1981 ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg
     2041 gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc
     2101 acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt
     2161 ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc
     2221 ttttgattta taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta
     2281 acaaaaattt aacgcgaatt ttaacaaaat attaacgctt acaatttcct gatgcggtat
     2341 tttctcctta cgcatctgtg cggtatttca caccgcatag atcggcaagt gcacaaacaa
     2401 tacttaaata aatactactc agtaataacc tatttcttag catttttgac gaaatttgct
     2461 attttgttag agtcttttac accatttgtc tccacacctc cgcttacatc aacaccaata
     2521 acgccattta atctaagcgc atcaccaaca ttttctggcg tcagtccacc agctaacata
     2581 aaatgtaagc tttcggggct ctcttgcctt ccaacccagt cagaaatcga gttccaatcc
     2641 aaaagttcac ctgtcccacc tgcttctgaa tcaaacaagg gaataaacga atgaggtttc
     2701 tgtgaagctg cactgagtag tatgttgcag tcttttggaa atacgagtct tttaataact
     2761 ggcaaaccga ggaactcttg gtattcttgc cacgactcat ctccatgcag ttggacgata
     2821 tcaatgccgt aatcattgac cagagccaaa acatcctcct taggttgatt acgaaacacg
     2881 ccaaccaagt atttcggagt gcctgaacta tttttatatg cttttacaag acttgaaatt
     2941 ttccttgcaa taaccgggtc aattgttctc tttctattgg gcacacatat aatacccagc
     3001 aagtcagcat cggaatctag agcacattct gcggcctctg tgctctgcaa gccgcaaact
     3061 ttcaccaatg gaccagaact acctgtgaaa ttaataacag acatactcca agctgccttt
     3121 gtgtgcttaa tcacgtatac tcacgtgctc aatagtcacc aatgccctcc ctcttggccc
     3181 tctccttttc ttttttcgac cgaattaatt cttaatcggc aaaaaaagaa aagctccgga
     3241 tcaagattgt acgtaaggtg acaagctatt tttcaataaa gaatatcttc cactactgcc
     3301 atctggcgtc ataactgcaa agtacacata tattacgatg ctgttctatt aaatgcttcc
     3361 tatattatat atatagtaat gtcgtgatct atggtgcact ctcagtacaa tctgctctga
     3421 tgccgcatag ttaagccagc cccgacaccc gccaacaccc gctgacgcgc cctgacgggc
     3481 ttgtctgctc ccggcatccg cttacagaca agctgtgacc gtctccggga gctgcatgtg
     3541 tcagaggttt tcaccgtcat caccgaaacg cgcgagacga aagggcctcg tgatacgcct
     3601 atttttatag gttaatgtca tgataataat ggtttcttag acggatcgct tgcctgtaac
     3661 ttacacgcgc ctcgtatctt ttaatgatgg aataatttgg gaatttactc tgtgtttatt
     3721 tatttttatg ttttgtattt ggattttaga aagtaaataa agaaggtaga agagttacgg
     3781 aatgaagaaa aaaaaataaa caaaggttta aaaaatttca acaaaaagcg tactttacat
     3841 atatatttat tagacaagaa aagcagatta aatagatata cattcgatta acgataagta
     3901 aaatgtaaaa tcacaggatt ttcgtgtgtg gtcttctaca cagacaagat gaaacaattc
     3961 ggcattaata cctgagagca ggaagagcaa gataaaaggt agtatttgtt ggcgatcccc
     4021 ctagagtctt ttacatcttc ggaaaacaaa aactattttt tctttaattt ctttttttac
     4081 tttctatttt taatttatat atttatatta aaaaatttaa attataatta tttttatagc
     4141 acgtgatgaa aaggacccag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg
     4201 tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat
     4261 gcttcaataa tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat
     4321 tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt
     4381 aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag
     4441 cggtaagatc cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa
     4501 agttctgcta tgtggcgcgg tattatcccg tattgacgcc gggcaagagc aactcggtcg
     4561 ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct
     4621 tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac
     4681 tgcggccaac ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca
     4741 caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat
     4801 accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact
     4861 attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc
     4921 ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga
     4981 taaatctgga gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg
     5041 taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg
     5101 aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca
     5161 agtttactca tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta
     5221 ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca
     5281 ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg
     5341 cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga
     5401 tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa
     5461 tactgttctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc
     5521 tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg
     5581 tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac
     5641 ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct
     5701 acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc
     5761 ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg
     5821 gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg
     5881 ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct
     5941 ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga
     6001 taaccgtatt accgcctttg agtgagctga taccgctcgc cgcagccgaa cgaccgagcg
     6061 cagcgagtca gtgagcgagg aagcggaaga gcgcccaata cgcaaaccgc ctctccccgc
     6121 gcgttggccg attcattaat gcagctggca cgacaggttt cccgactgga aagcgggcag
     6181 tgagcgcaac gcaattaatg tgagttagct cactcattag gcaccccagg ctttacactt
     6241 tatgcttccg gctcgtatgt tgtgtgga
//
