LOCUS       sequence-441270-        6559 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Yeast optimized human alpha synuclein fused with aga2 protein gene 
            under gal1 promoter for the expression of alpha synuclein in yeast 
            surface display..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6559)
  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 6559)
  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-alpha-Synuclein-c_myc
FEATURES             Location/Qualifiers
     source          1..6559
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      63..518
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(150..169)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     360..381
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(621..1295)
                     /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(1296..1576)
                     /gene="S. cerevisiae TRP1"
                     /label=TRP1 promoter
     primer_bind     complement(1597..1616)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     1716..1738
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(1776..1794)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     misc_feature    1835..2338
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     promoter        2375..2479
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2480..2548,2549..3340)
                     /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(2698..2717)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3511..4099
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4000..4019
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4253..4270
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    4387..4408
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(4423..4428,4429..4446,4447..4453)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4461..4477
                     /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     4466..4488
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     4485..4501
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4485..4501
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        4522..4540
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        4570..5222
                     /gene="GAL1 and GAL10 genes of S. cerevisiae"
                     /label=GAL1,10 promoter
                     /note="divergent inducible promoter, regulated by Gal4"
     protein_bind    4774..4891
                     /label=UAS
                     /bound_moiety="Gal4"
                     /note="upstream activating sequence mediating 
                     Gal4-dependent induction"
     primer_bind     5186..5209
                     /label=GAL1
                     /note="S. cerevisiae GAL1 promoter, forward primer"
     CDS             5541..5552
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     CDS             5553..5579
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             6081..6110
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     primer_bind     complement(6439..6458)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(6440..6458)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(6465..6482)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(6465..6481)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6474..6496)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
ORIGIN
        1 atagcgaaga ggcccgcacc gatcgccttt cccaacagtt gcgcagcctg aatggcgaat
       61 ggacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac
      121 cgctacactt gccagcgccc tagcgcccgc tcctttcgct ttcttccctt cctttctcgc
      181 cacgttcgcc ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt
      241 tagtgcttta cggcacctcg accccaaaaa acttgattag ggtgatggtt cacgtagtgg
      301 gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag
      361 tggactcttg ttccaaactg gaacaacact caaccctatc tcggtctatt cttttgattt
      421 ataagggatt ttgccgattt cggcctattg gttaaaaaat gagctgattt aacaaaaatt
      481 taacgcgaat tttaacaaaa tattaacgct tacaatttcc tgatgcggta ttttctcctt
      541 acgcatctgt gcggtatttc acaccgcata gatcggcaag tgcacaaaca atacttaaat
      601 aaatactact cagtaataac ctatttctta gcatttttga cgaaatttgc tattttgtta
      661 gagtctttta caccatttgt ctccacacct ccgcttacat caacaccaat aacgccattt
      721 aatctaagcg catcaccaac attttctggc gtcagtccac cagctaacat aaaatgtaag
      781 ctttcggggc tctcttgcct tccaacccag tcagaaatcg agttccaatc caaaagttca
      841 cctgtcccac ctgcttctga atcaaacaag ggaataaacg aatgaggttt ctgtgaagct
      901 gcactgagta gtatgttgca gtcttttgga aatacgagtc ttttaataac tggcaaaccg
      961 aggaactctt ggtattcttg ccacgactca tctccatgca gttggacgat atcaatgccg
     1021 taatcattga ccagagccaa aacatcctcc ttaggttgat tacgaaacac gccaaccaag
     1081 tatttcggag tgcctgaact atttttatat gcttttacaa gacttgaaat tttccttgca
     1141 ataaccgggt caattgttct ctttctattg ggcacacata taatacccag caagtcagca
     1201 tcggaatcta gagcacattc tgcggcctct gtgctctgca agccgcaaac tttcaccaat
     1261 ggaccagaac tacctgtgaa attaataaca gacatactcc aagctgcctt tgtgtgctta
     1321 atcacgtata ctcacgtgct caatagtcac caatgccctc cctcttggcc ctctcctttt
     1381 cttttttcga ccgaattaat tcttaatcgg caaaaaaaga aaagctccgg atcaagattg
     1441 tacgtaaggt gacaagctat ttttcaataa agaatatctt ccactactgc catctggcgt
     1501 cataactgca aagtacacat atattacgat gctgttctat taaatgcttc ctatattata
     1561 tatatagtaa tgtcgtgatc tatggtgcac tctcagtaca atctgctctg atgccgcata
     1621 gttaagccag ccccgacacc cgccaacacc cgctgacgcg ccctgacggg cttgtctgct
     1681 cccggcatcc gcttacagac aagctgtgac cgtctccggg agctgcatgt gtcagaggtt
     1741 ttcaccgtca tcaccgaaac gcgcgagacg aaagggcctc gtgatacgcc tatttttata
     1801 ggttaatgtc atgataataa tggtttctta gacggatcgc ttgcctgtaa cttacacgcg
     1861 cctcgtatct tttaatgatg gaataatttg ggaatttact ctgtgtttat ttatttttat
     1921 gttttgtatt tggattttag aaagtaaata aagaaggtag aagagttacg gaatgaagaa
     1981 aaaaaaataa acaaaggttt aaaaaatttc aacaaaaagc gtactttaca tatatattta
     2041 ttagacaaga aaagcagatt aaatagatat acattcgatt aacgataagt aaaatgtaaa
     2101 atcacaggat tttcgtgtgt ggtcttctac acagacaaga tgaaacaatt cggcattaat
     2161 acctgagagc aggaagagca agataaaagg tagtatttgt tggcgatccc cctagagtct
     2221 tttacatctt cggaaaacaa aaactatttt ttctttaatt tcttttttta ctttctattt
     2281 ttaatttata tatttatatt aaaaaattta aattataatt atttttatag cacgtgatga
     2341 aaaggaccca ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt
     2401 ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata
     2461 atattgaaaa aggaagagta tgagtattca acatttccgt gtcgccctta ttcccttttt
     2521 tgcggcattt tgccttcctg tttttgctca cccagaaacg ctggtgaaag taaaagatgc
     2581 tgaagatcag ttgggtgcac gagtgggtta catcgaactg gatctcaaca gcggtaagat
     2641 ccttgagagt tttcgccccg aagaacgttt tccaatgatg agcactttta aagttctgct
     2701 atgtggcgcg gtattatccc gtattgacgc cgggcaagag caactcggtc gccgcataca
     2761 ctattctcag aatgacttgg ttgagtactc accagtcaca gaaaagcatc ttacggatgg
     2821 catgacagta agagaattat gcagtgctgc cataaccatg agtgataaca ctgcggccaa
     2881 cttacttctg acaacgatcg gaggaccgaa ggagctaacc gcttttttgc acaacatggg
     2941 ggatcatgta actcgccttg atcgttggga accggagctg aatgaagcca taccaaacga
     3001 cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg
     3061 cgaactactt actctagctt cccggcaaca attaatagac tggatggagg cggataaagt
     3121 tgcaggacca cttctgcgct cggcccttcc ggctggctgg tttattgctg ataaatctgg
     3181 agccggtgag cgtgggtctc gcggtatcat tgcagcactg gggccagatg gtaagccctc
     3241 ccgtatcgta gttatctaca cgacggggag tcaggcaact atggatgaac gaaatagaca
     3301 gatcgctgag ataggtgcct cactgattaa gcattggtaa ctgtcagacc aagtttactc
     3361 atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat
     3421 cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc
     3481 agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg
     3541 ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct
     3601 accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct
     3661 tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct
     3721 cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg
     3781 gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc
     3841 gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga
     3901 gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg
     3961 cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta
     4021 tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg
     4081 ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg
     4141 ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat
     4201 taccgccttt gagtgagctg ataccgctcg ccgcagccga acgaccgagc gcagcgagtc
     4261 agtgagcgag gaagcggaag agcgcccaat acgcaaaccg cctctccccg cgcgttggcc
     4321 gattcattaa tgcagctggc acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa
     4381 cgcaattaat gtgagttagc tcactcatta ggcaccccag gctttacact ttatgcttcc
     4441 ggctcgtatg ttgtgtggaa ttgtgagcgg ataacaattt cacacaggaa acagctatga
     4501 ccatgattac gccaagctcg aaattaaccc tcactaaagg gaacaaaagc tggtaccaat
     4561 tccttgaatt ttcaaaaatt cttacttttt ttttggatgg acgcaaagaa gtttaataat
     4621 catattacat ggcattacca ccatatacat atccatatct aatcttactt atatgttgtg
     4681 gaaatgtaaa gagccccatt atcttagcct aaaaaaacct tctctttgga actttcagta
     4741 atacgcttaa ctgctcattg ctatattgaa gtacggatta gaagccgccg agcgggtgac
     4801 agccctccga aggaagactc tcctccgtgc gtcctcgtct tcaccggtcg cgttcctgaa
     4861 acgcagatgt gcctcgcgcc gcactgctcc gaacaataaa gattctacaa tactagcttt
     4921 tatggttatg aagaggaaaa attggcagta acctggcccc acaaaccttc aaatgaacga
     4981 atcaaattaa caaccatagg atgataatgc gattagtttt ttagccttat ttctggggta
     5041 attaatcagc gaagcgatga tttttgatct attaacagat atataaatgc aaaaactgca
     5101 taaccacttt aactaatact ttcaacattt tcggtttgta ttacttctta ttcaaatgta
     5161 ataaaagtat caacaaaaaa ttgttaatat acctctatac tttaacgtca aggagaaaaa
     5221 accccggatc gaattcccta cttcatacat tttcaattaa gatgcagtta cttcgctgtt
     5281 tttcaatatt ttctgttatt gcttcagttt tagcacagga actgacaact atatgcgagc
     5341 aaatcccctc accaacttta gaatcgacgc cgtactcttt gtcaacgact actattttgg
     5401 ccaacgggaa ggcaatgcaa ggagtttttg aatattacaa atcagtaacg tttgtcagta
     5461 attgcggttc tcacccctca acaactagca aaggcagccc cataaacaca cagtatgttt
     5521 ttaaggacaa tagctcgacg attgaaggta gatacccata cgacgttcca gactacgctc
     5581 tgcaggctag tggtggagga ggctctggtg gaggcggtag cggaggcgga gggtcgacta
     5641 gtatggacgt tttcatgaag ggtctttcta aagcgaaaga gggcgtggta gctgcggccg
     5701 aaaaaactaa acaaggggtg gccgaggctg ctgggaaaac gaaggaaggt gtattgtacg
     5761 ttggttcaaa gaccaaagag ggagtagttc acggagtcgc cacagttgcc gagaagacca
     5821 aggaacaggt aacgaatgtg ggaggtgcag tggtgactgg tgtcactgcg gtcgcccaaa
     5881 aaacagttga aggagcggga tcaatagccg cagcaacggg atttgttaag aaggaccaat
     5941 taggaaaaaa tgaagaggga gcacctcaag aaggtattct agaggatatg ccagtcgacc
     6001 ccgataacga ggcttatgag atgccgtcag aggaagggta tcaggactat gagccagaag
     6061 ccctcgaggg gggcggatcc gaacaaaagc ttatttctga agaggacttg taatagagat
     6121 ctgataacaa cagtgtagat gtaacaaaat cgactttgtt cccactgtac ttttagctcg
     6181 tacaaaatac aatatacttt tcatttctcc gtaaacaaca tgttttccca tgtaatatcc
     6241 ttttctattt ttcgttccgt taccaacttt acacatactt tatatagcta ttcacttcta
     6301 tacactaaaa aactaagaca attttaattt tgctgcctgc catatttcaa tttgttataa
     6361 attcctataa tttatcctat tagtagctaa aaaaagatga atgtgaatcg aatcctaaga
     6421 gaattgagct ccaattcgcc ctatagtgag tcgtattaca attcactggc cgtcgtttta
     6481 caacgtcgtg actgggaaaa ccctggcgtt acccaactta atcgccttgc agcacatccc
     6541 cctttcgcca gctggcgta
//
