LOCUS       sequence-460503-        6502 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Alpha synuclein gene fused with gst gene under t7 promoter for 
            bacterial expression of alpha synuclein protein..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6502)
  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 6502)
  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: pET22b-T7-6h-GST-alpha-Synuclein
FEATURES             Location/Qualifiers
     source          1..6502
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     121..140
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     224..243
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        224..242
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    243..267
                     /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)."
     CDS             311..328
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             338..991
                     /codon_start=1
                     /product="glutathione S-transferase from Schistosoma 
                     japonicum"
                     /label=GST
                     /translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
                     FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
                     GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
                     YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
     CDS             998..1018
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             1457..1474
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(1527..1545)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1541..1588
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      1625..2080
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1712..1731)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1922..1943
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        2106..2210
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2211..2279,2280..3071)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(2429..2448)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3242..3830
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3731..3750
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3984..4001
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    4016..4155
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(4081..4100)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4241..4263
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(4257..4448)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(5257..6339)
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor"
                     /label=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)."
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     primer_bind     6301..6320
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(6340..6417)
                     /gene="lacI"
                     /label=lacI promoter
ORIGIN
        1 aaggaatggt gcatgcaagg agatggcgcc caacagtccc ccggccacgg ggcctgccac
       61 catacccacg ccgaaacaag cgctcatgag cccgaagtgg cgagcccgat cttccccatc
      121 ggtgatgtcg gcgatatagg cgccagcaac cgcacctgtg gcgccggtga tgccggccac
      181 gatgcgtccg gcgtagagga tcgagatctc gatcccgcga aattaatacg actcactata
      241 ggggaattgt gagcggataa caattcccct ctagattaac tttaagaagg agggtaccat
      301 gaacaccatt catcaccatc accatcacaa cactagtatg tcccctatac taggttattg
      361 gaaaattaag ggccttgtgc aacccactcg acttcttttg gaatatcttg aagaaaaata
      421 tgaagagcat ttgtatgagc gcgatgaagg tgataaatgg cgaaacaaaa agtttgaatt
      481 gggtttggag tttcccaatc ttccttatta tattgatggt gatgttaaat taacacagtc
      541 tatggccatc atacgttata tagctgacaa gcacaacatg ttgggtggtt gtccaaaaga
      601 gcgtgcagag atttcaatgc ttgaaggagc ggttttggat attagatacg gtgtttcgag
      661 aattgcatat agtaaagact ttgaaactct caaagttgat tttcttagca agctacctga
      721 aatgctgaaa atgttcgaag atcgtttatg tcataaaaca tatttaaatg gtgatcatgt
      781 aacccatcct gacttcatgt tgtatgacgc tcttgatgtt gttttataca tggacccaat
      841 gtgcctggat gcgttcccaa aattagtttg ttttaaaaaa cgtattgaag ctatcccaca
      901 aattgataag tacttgaaat ccagcaagta tatagcatgg cctttgcagg gctggcaagc
      961 cacgtttggt ggtggcgacc atcctccaaa aggatccgaa aacctgtatt ttcagggcac
     1021 tagtatggac gttttcatga agggtctttc taaagcgaaa gagggcgtgg tagctgcggc
     1081 cgaaaaaact aaacaagggg tggccgaggc tgctgggaaa acgaaggaag gtgtattgta
     1141 cgttggttca aagaccaaag agggagtagt tcacggagtc gccacagttg ccgagaagac
     1201 caaggaacag gtaacgaatg tgggaggtgc agtggtgact ggtgtcactg cggtcgccca
     1261 aaaaacagtt gaaggagcgg gatcaatagc cgcagcaacg ggatttgtta agaaggacca
     1321 attaggaaaa aatgaagagg gagcacctca agaaggtatt ctagaggata tgccagtcga
     1381 ccccgataac gaggcttatg agatgccgtc agaggaaggg tatcaggact atgagccaga
     1441 agcctgatga ctcgagcacc accaccacca ccactgagat ccggctgcta acaaagcccg
     1501 aaaggaagct gagttggctg ctgccaccgc tgagcaataa ctagcataac cccttggggc
     1561 ctctaaacgg gtcttgaggg gttttttgct gaaaggagga actatatccg gattggcgaa
     1621 tgggacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg
     1681 accgctacac ttgccagcgc cctagcgccc gctcctttcg ctttcttccc ttcctttctc
     1741 gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt agggttccga
     1801 tttagtgctt tacggcacct cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt
     1861 gggccatcgc cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat
     1921 agtggactct tgttccaaac tggaacaaca ctcaacccta tctcggtcta ttcttttgat
     1981 ttataaggga ttttgccgat ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa
     2041 tttaacgcga attttaacaa aatattaacg cttacaattt aggtggcact tttcggggaa
     2101 atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca
     2161 tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt atgagtattc
     2221 aacatttccg tgtcgccctt attccctttt ttgcggcatt ttgccttcct gtttttgctc
     2281 acccagaaac gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt
     2341 acatcgaact ggatctcaac agcggtaaga tccttgagag ttttcgcccc gaagaacgtt
     2401 ttccaatgat gagcactttt aaagttctgc tatgtggcgc ggtattatcc cgtattgacg
     2461 ccgggcaaga gcaactcggt cgccgcatac actattctca gaatgacttg gttgagtact
     2521 caccagtcac agaaaagcat cttacggatg gcatgacagt aagagaatta tgcagtgctg
     2581 ccataaccat gagtgataac actgcggcca acttacttct gacaacgatc ggaggaccga
     2641 aggagctaac cgcttttttg cacaacatgg gggatcatgt aactcgcctt gatcgttggg
     2701 aaccggagct gaatgaagcc ataccaaacg acgagcgtga caccacgatg cctgcagcaa
     2761 tggcaacaac gttgcgcaaa ctattaactg gcgaactact tactctagct tcccggcaac
     2821 aattaataga ctggatggag gcggataaag ttgcaggacc acttctgcgc tcggcccttc
     2881 cggctggctg gtttattgct gataaatctg gagccggtga gcgtgggtct cgcggtatca
     2941 ttgcagcact ggggccagat ggtaagccct cccgtatcgt agttatctac acgacgggga
     3001 gtcaggcaac tatggatgaa cgaaatagac agatcgctga gataggtgcc tcactgatta
     3061 agcattggta actgtcagac caagtttact catatatact ttagattgat ttaaaacttc
     3121 atttttaatt taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc
     3181 cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt
     3241 cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac
     3301 cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct
     3361 tcagcagagc gcagatacca aatactgtcc ttctagtgta gccgtagtta ggccaccact
     3421 tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg
     3481 ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata
     3541 aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga
     3601 cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag
     3661 ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg
     3721 agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac
     3781 ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca
     3841 acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg ttctttcctg
     3901 cgttatcccc tgattctgtg gataaccgta ttaccgcctt tgagtgagct gataccgctc
     3961 gccgcagccg aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa gagcgcctga
     4021 tgcggtattt tctccttacg catctgtgcg gtatttcaca ccgcaatggt gcactctcag
     4081 tacaatctgc tctgatgccg catagttaag ccagtataca ctccgctatc gctacgtgac
     4141 tgggtcatgg ctgcgccccg acacccgcca acacccgctg acgcgccctg acgggcttgt
     4201 ctgctcccgg catccgctta cagacaagct gtgaccgtct ccgggagctg catgtgtcag
     4261 aggttttcac cgtcatcacc gaaacgcgcg aggcagctgc ggtaaagctc atcagcgtgg
     4321 tcgtgaagcg attcacagat gtctgcctgt tcatccgcgt ccagctcgtt gagtttctcc
     4381 agaagcgtta atgtctggct tctgataaag cgggccatgt taagggcggt tttttcctgt
     4441 ttggtcactg atgcctccgt gtaaggggga tttctgttca tgggggtaat gataccgatg
     4501 aaacgagaga ggatgctcac gatacgggtt actgatgatg aacatgcccg gttactggaa
     4561 cgttgtgagg gtaaacaact ggcggtatgg atgcggcggg accagagaaa aatcactcag
     4621 ggtcaatgcc agcgcttcgt taatacagat gtaggtgttc cacagggtag ccagcagcat
     4681 cctgcgatgc agatccggaa cataatggtg cagggcgctg acttccgcgt ttccagactt
     4741 tacgaaacac ggaaaccgaa gaccattcat gttgttgctc aggtcgcaga cgttttgcag
     4801 cagcagtcgc ttcacgttcg ctcgcgtatc ggtgattcat tctgctaacc agtaaggcaa
     4861 ccccgccagc ctagccgggt cctcaacgac aggagcacga tcatgcgcac ccgtggggcc
     4921 gccatgccgg cgataatggc ctgcttctcg ccgaaacgtt tggtggcggg accagtgacg
     4981 aaggcttgag cgagggcgtg caagattccg aataccgcaa gcgacaggcc gatcatcgtc
     5041 gcgctccagc gaaagcggtc ctcgccgaaa atgacccaga gcgctgccgg cacctgtcct
     5101 acgagttgca tgataaagaa gacagtcata agtgcggcga cgatagtcat gccccgcgcc
     5161 caccggaagg agctgactgg gttgaaggct ctcaagggca tcggtcgaga tcccggtgcc
     5221 taatgagtga gctaacttac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga
     5281 aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt
     5341 attgggcgcc agggtggttt ttcttttcac cagtgagacg ggcaacagct gattgccctt
     5401 caccgcctgg ccctgagaga gttgcagcaa gcggtccacg ctggtttgcc ccagcaggcg
     5461 aaaatcctgt ttgatggtgg ttaacggcgg gatataacat gagctgtctt cggtatcgtc
     5521 gtatcccact accgagatat ccgcaccaac gcgcagcccg gactcggtaa tggcgcgcat
     5581 tgcgcccagc gccatctgat cgttggcaac cagcatcgca gtgggaacga tgccctcatt
     5641 cagcatttgc atggtttgtt gaaaaccgga catggcactc cagtcgcctt cccgttccgc
     5701 tatcggctga atttgattgc gagtgagata tttatgccag ccagccagac gcagacgcgc
     5761 cgagacagaa cttaatgggc ccgctaacag cgcgatttgc tggtgaccca atgcgaccag
     5821 atgctccacg cccagtcgcg taccgtcttc atgggagaaa ataatactgt tgatgggtgt
     5881 ctggtcagag acatcaagaa ataacgccgg aacattagtg caggcagctt ccacagcaat
     5941 ggcatcctgg tcatccagcg gatagttaat gatcagccca ctgacgcgtt gcgcgagaag
     6001 attgtgcacc gccgctttac aggcttcgac gccgcttcgt tctaccatcg acaccaccac
     6061 gctggcaccc agttgatcgg cgcgagattt aatcgccgcg acaatttgcg acggcgcgtg
     6121 cagggccaga ctggaggtgg caacgccaat cagcaacgac tgtttgcccg ccagttgttg
     6181 tgccacgcgg ttgggaatgt aattcagctc cgccatcgcc gcttccactt tttcccgcgt
     6241 tttcgcagaa acgtggctgg cctggttcac cacgcgggaa acggtctgat aagagacacc
     6301 ggcatactct gcgacatcgt ataacgttac tggtttcaca ttcaccaccc tgaattgact
     6361 ctcttccggg cgctatcatg ccataccgcg aaaggttttg cgccattcga tggtgtccgg
     6421 gatctcgacg ctctccctta tgcgactcct gcattaggaa gcagcccagt agtaggttga
     6481 ggccgttgag caccgccgcc gc
//
