LOCUS       sequence-207256-        6123 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial expression of mutant human alpha synuclein.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6123)
  TITLE     Lashuel Lab plasmids
REFERENCE   2  (bases 1 to 6123)
  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"
COMMENT     Sequence Label: pET11a asyn Q79E (TAC-->TAT)
FEATURES             Location/Qualifiers
     source          1..6123
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      32..619
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     520..539
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     773..790
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    805..944
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(870..889)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     1030..1052
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(1046..1237)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(2549..3631)
                     /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     3593..3612
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(3632..3709)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     3915..3934
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     4018..4037
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        4018..4036
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    4037..4061
                     /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)."
     RBS             4076..4098
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             4532..4564
                     /codon_start=1
                     /product="leader peptide from bacteriophage T7 gene 10"
                     /label=T7 tag (gene 10 leader)
                     /note="promotes efficient translation in E. coli"
                     /translation="MASMTGGQQMG"
     primer_bind     complement(4642..4660)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      4656..4703
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        complement(join(4878..4883,4884..4900,4901..4906))
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     primer_bind     complement(4933..4951)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5019..5123
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5124..5192,5193..5984)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(5342..5361)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct
       61 gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc
      121 taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgtcc
      181 ttctagtgta gccgtagtta ggccaccact tcaagaacct gtagcaccgc ctacatacct
      241 cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg
      301 gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc
      361 gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga
      421 gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg
      481 cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta
      541 tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg
      601 ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg
      661 ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat
      721 taccgccttt gagtgagctg ataccgctcg ccgcagccga acgaccgagc gcagcgagtc
      781 agtgagcgag gaagcggaag agcgcctgat gcggtatttt ctccttacgc atctgtgcgg
      841 tatttcacac cgcaatggtg cactctcagt acaatctgct ctgatgccgc atagttaagc
      901 cagtatacac tccgctatcg ctacgtgact gggtcatggc tgcgccccga cacccgccaa
      961 cacccgctga cgcgccctga cgggcttgtc tgctcccggc atccgcttac agacaagctg
     1021 tgaccgtctc cgggagctgc atgtgtcaga ggttttcacc gtcatcaccg aaacgcgcga
     1081 ggcagctgcg gtaaagctca tcagcgtggt cgtgaagcga ttcacagatg tctgcctgtt
     1141 catccgcgtc cagctcgttg agtttctcca gaagcgttaa tgtctggctt ctgataaagc
     1201 gggccatgtt aagggcggtt ttttcctgtt tggtcactga tgcctccgtg taagggggat
     1261 ttctgttcat gggggtaatg ataccgatga aacgagagag gatgctcacg atacgggtta
     1321 ctgatgatga acatgcccgg ttactggaac gttgtgaggg taaacaactg gcggtatgga
     1381 tgcggcggga ccagagaaaa atcactcagg gtcaatgcca gcgcttcgtt aatacagatg
     1441 taggtgttcc acagggtagc cagcagcatc ctgcgatgca gatccggaac ataatggtgc
     1501 agggcgctga cttccgcgtt tccagacttt acgaaacacg gaaaccgaag accattcatg
     1561 ttgttgctca ggtcgcagac gttttgcagc agcagtcgct tcacgttcgc tcgcgtatcg
     1621 gtgattcatt ctgctaacca gtaaggcaac cccgccagcc tagccgggtc ctcaacgaca
     1681 ggagcacgat catgcgcacc cgtggccagg acccaacgct gcccgagatg cgccgcgtgc
     1741 ggctgctgga gatggcggac gcgatggata tgttctgcca agggttggtt tgcgcattca
     1801 cagttctccg caagaattga ttggctccaa ttcttggagt ggtgaatccg ttagcgaggt
     1861 gccgccggct tccattcagg tcgaggtggc ccggctccat gcaccgcgac gcaacgcggg
     1921 gaggcagaca aggtataggg cggcgcctac aatccatgcc aacccgttcc atgtgctcgc
     1981 cgaggcggca taaatcgccg tgacgatcag cggtccaatg atcgaagtta ggctggtaag
     2041 agccgcgagc gatccttgaa gctgtccctg atggtcgtca tctacctgcc tggacagcat
     2101 ggcctgcaac gcgggcatcc cgatgccgcc ggaagcgaga agaatcataa tggggaaggc
     2161 catccagcct cgcgtcgcga acgccagcaa gacgtagccc agcgcgtcgg ccgccatgcc
     2221 ggcgataatg gcctgcttct cgccgaaacg tttggtggcg ggaccagtga cgaaggcttg
     2281 agcgagggcg tgcaagattc cgaataccgc aagcgacagg ccgatcatcg tcgcgctcca
     2341 gcgaaagcgg tcctcgccga aaatgaccca gagcgctgcc ggcacctgtc ctacgagttg
     2401 catgataaag aagacagtca taagtgcggc gacgatagtc atgccccgcg cccaccggaa
     2461 ggagctgact gggttgaagg ctctcaaggg catcggtcga gatcccggtg cctaatgagt
     2521 gagctaactt acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc
     2581 gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg
     2641 ccagggtggt ttttcttttc accagtgaga cgggcaacag ctgattgccc ttcaccgcct
     2701 ggccctgaga gagttgcagc aagcggtcca cgctggtttg ccccagcagg cgaaaatcct
     2761 gtttgatggt ggttaacggc gggatataac atgagctgtc ttcggtatcg tcgtatccca
     2821 ctaccgagat atccgcacca acgcgcagcc cggactcggt aatggcgcgc attgcgccca
     2881 gcgccatctg atcgttggca accagcatcg cagtgggaac gatgccctca ttcagcattt
     2941 gcatggtttg ttgaaaaccg gacatggcac tccagtcgcc ttcccgttcc gctatcggct
     3001 gaatttgatt gcgagtgaga tatttatgcc agccagccag acgcagacgc gccgagacag
     3061 aacttaatgg gcccgctaac agcgcgattt gctggtgacc caatgcgacc agatgctcca
     3121 cgcccagtcg cgtaccgtct tcatgggaga aaataatact gttgatgggt gtctggtcag
     3181 agacatcaag aaataacgcc ggaacattag tgcaggcagc ttccacagca atggcatcct
     3241 ggtcatccag cggatagtta atgatcagcc cactgacgcg ttgcgcgaga agattgtgca
     3301 ccgccgcttt acaggcttcg acgccgcttc gttctaccat cgacaccacc acgctggcac
     3361 ccagttgatc ggcgcgagat ttaatcgccg cgacaatttg cgacggcgcg tgcagggcca
     3421 gactggaggt ggcaacgcca atcagcaacg actgtttgcc cgccagttgt tgtgccacgc
     3481 ggttgggaat gtaattcagc tccgccatcg ccgcttccac tttttcccgc gttttcgcag
     3541 aaacgtggct ggcctggttc accacgcggg aaacggtctg ataagagaca ccggcatact
     3601 ctgcgacatc gtataacgtt actggtttca cattcaccac cctgaattga ctctcttccg
     3661 ggcgctatca tgccataccg cgaaaggttt tgcgccattc gatggtgtcc gggatctcga
     3721 cgctctccct tatgcgactc ctgcattagg aagcagccca gtagtaggtt gaggccgttg
     3781 agcaccgccg ccgcaaggaa tggtgcatgc aaggagatgg cgcccaacag tcccccggcc
     3841 acggggcctg ccaccatacc cacgccgaaa caagcgctca tgagcccgaa gtggcgagcc
     3901 cgatcttccc catcggtgat gtcggcgata taggcgccag caaccgcacc tgtggcgccg
     3961 gtgatgccgg ccacgatgcg tccggcgtag aggatcgaga tctcgatccc gcgaaattaa
     4021 tacgactcac tataggggaa ttgtgagcgg ataacaattc ccctctagaa ataattttgt
     4081 ttaactttaa gaaggagata tacatatgga tgtattcatg aaaggacttt caaaggccaa
     4141 ggagggagtt gtggctgctg ctgagaaaac caaacagggt gtggcagaag cagcaggaaa
     4201 gacaaaagag ggtgttctct atgtaggctc caaaaccaag gagggagtgg tgcatggtgt
     4261 ggcaacagtg gctgagaaga ccaaagagca agtgacaaat gttggaggag cagtggtgac
     4321 gggtgtgaca gcagtagccg agaagacagt ggagggagcg gggagcattg cagcagccac
     4381 tggctttgtc aaaaaggacc agttgggcaa gaatgaagaa ggagccccac aggaaggaat
     4441 tctggaagat atgcctgtgg atcctgacaa tgaggcttat gaaatgcctt ctgaggaagg
     4501 gtatcaagac tatgaacctg aagcctaaca tatggctagc atgactggtg gacagcaaat
     4561 gggtcgcgga tcccgggtac ctcgagtcga ccgatccggc tgctaacaaa gcccgaaagg
     4621 aagctgagtt ggctgctgcc accgctgagc aataactagc ataacccctt ggggcctcta
     4681 aacgggtctt gaggggtttt ttgctgaaag gaggaactat atccggatat cccgcaagag
     4741 gcccggcagt accggcataa ccaagcctat gcctacagca tccagggtga cggtgccgag
     4801 gatgacgatg agcgcattgt tagatttcat acacggtgcc tgactgcgtt agcaatttaa
     4861 ctgtgataaa ctaccgcatt aaagcttatc gatgataagc tgtcaaacat gagaattctt
     4921 gaagacgaaa gggcctcgtg atacgcctat ttttataggt taatgtcatg ataataatgg
     4981 tttcttagac gtcaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat
     5041 ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc
     5101 aataatattg aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc cttattccct
     5161 tttttgcggc attttgcctt cctgtttttg ctcacccaga aacgctggtg aaagtaaaag
     5221 atgctgaaga tcagttgggt gcacgagtgg gttacatcga actggatctc aacagcggta
     5281 agatccttga gagttttcgc cccgaagaac gttttccaat gatgagcact tttaaagttc
     5341 tgctatgtgg cgcggtatta tcccgtgttg acgccgggca agagcaactc ggtcgccgca
     5401 tacactattc tcagaatgac ttggttgagt actcaccagt cacagaaaag catcttacgg
     5461 atggcatgac agtaagagaa ttatgcagtg ctgccataac catgagtgat aacactgcgg
     5521 ccaacttact tctgacaacg atcggaggac cgaaggagct aaccgctttt ttgcacaaca
     5581 tgggggatca tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa gccataccaa
     5641 acgacgagcg tgacaccacg atgcctgcag caatggcaac aacgttgcgc aaactattaa
     5701 ctggcgaact acttactcta gcttcccggc aacaattaat agactggatg gaggcggata
     5761 aagttgcagg accacttctg cgctcggccc ttccggctgg ctggtttatt gctgataaat
     5821 ctggagccgg tgagcgtggg tctcgcggta tcattgcagc actggggcca gatggtaagc
     5881 cctcccgtat cgtagttatc tacacgacgg ggagtcaggc aactatggat gaacgaaata
     5941 gacagatcgc tgagataggt gcctcactga ttaagcattg gtaactgtca gaccaagttt
     6001 actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga
     6061 agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag
     6121 cgt
//
