LOCUS       sequence-269806-        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 Q99E (TAC-->TAT)
FEATURES             Location/Qualifiers
     source          1..6123
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             193..225
                     /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(303..321)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      317..364
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        complement(join(539..544,545..561,562..567))
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     primer_bind     complement(594..612)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        680..784
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(785..853,854..1645)
                     /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(1003..1022)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1816..2403
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2304..2323
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2557..2574
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    2589..2728
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(2654..2673)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2814..2836
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(2830..3021)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(4333..5415)
                     /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     5377..5396
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(5416..5493)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     5699..5718
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     5802..5821
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        5802..5820
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    5821..5845
                     /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             5860..5882
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
ORIGIN
        1 cagaagacag tggagggagc ggggagcatt gcagcagcca ctggctttgt caaaaaggac
       61 gagttgggca agaatgaaga aggagcccca caggaaggaa ttctggaaga tatgcctgtg
      121 gatcctgaca atgaggctta tgaaatgcct tctgaggaag ggtatcaaga ctatgaacct
      181 gaagcctaac atatggctag catgactggt ggacagcaaa tgggtcgcgg atcccgggta
      241 cctcgagtcg accgatccgg ctgctaacaa agcccgaaag gaagctgagt tggctgctgc
      301 caccgctgag caataactag cataacccct tggggcctct aaacgggtct tgaggggttt
      361 tttgctgaaa ggaggaacta tatccggata tcccgcaaga ggcccggcag taccggcata
      421 accaagccta tgcctacagc atccagggtg acggtgccga ggatgacgat gagcgcattg
      481 ttagatttca tacacggtgc ctgactgcgt tagcaattta actgtgataa actaccgcat
      541 taaagcttat cgatgataag ctgtcaaaca tgagaattct tgaagacgaa agggcctcgt
      601 gatacgccta tttttatagg ttaatgtcat gataataatg gtttcttaga cgtcaggtgg
      661 cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa
      721 tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa
      781 gagtatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct
      841 tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg
      901 tgcacgagtg ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg
      961 ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt
     1021 atcccgtgtt gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga
     1081 cttggttgag tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga
     1141 attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac
     1201 gatcggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg
     1261 ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac
     1321 gatgcctgca gcaatggcaa caacgttgcg caaactatta actggcgaac tacttactct
     1381 agcttcccgg caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct
     1441 gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg
     1501 gtctcgcggt atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat
     1561 ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg
     1621 tgcctcactg attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat
     1681 tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct
     1741 catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa
     1801 gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa
     1861 aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc
     1921 gaaggtaact ggcttcagca gagcgcagat accaaatact gtccttctag tgtagccgta
     1981 gttaggccac cacttcaaga acctgtagca ccgcctacat acctcgctct gctaatcctg
     2041 ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga
     2101 tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc
     2161 ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc
     2221 acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga
     2281 gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt
     2341 cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg
     2401 aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac
     2461 atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc ctttgagtga
     2521 gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg
     2581 gaagagcgcc tgatgcggta ttttctcctt acgcatctgt gcggtatttc acaccgcaat
     2641 ggtgcactct cagtacaatc tgctctgatg ccgcatagtt aagccagtat acactccgct
     2701 atcgctacgt gactgggtca tggctgcgcc ccgacacccg ccaacacccg ctgacgcgcc
     2761 ctgacgggct tgtctgctcc cggcatccgc ttacagacaa gctgtgaccg tctccgggag
     2821 ctgcatgtgt cagaggtttt caccgtcatc accgaaacgc gcgaggcagc tgcggtaaag
     2881 ctcatcagcg tggtcgtgaa gcgattcaca gatgtctgcc tgttcatccg cgtccagctc
     2941 gttgagtttc tccagaagcg ttaatgtctg gcttctgata aagcgggcca tgttaagggc
     3001 ggttttttcc tgtttggtca ctgatgcctc cgtgtaaggg ggatttctgt tcatgggggt
     3061 aatgataccg atgaaacgag agaggatgct cacgatacgg gttactgatg atgaacatgc
     3121 ccggttactg gaacgttgtg agggtaaaca actggcggta tggatgcggc gggaccagag
     3181 aaaaatcact cagggtcaat gccagcgctt cgttaataca gatgtaggtg ttccacaggg
     3241 tagccagcag catcctgcga tgcagatccg gaacataatg gtgcagggcg ctgacttccg
     3301 cgtttccaga ctttacgaaa cacggaaacc gaagaccatt catgttgttg ctcaggtcgc
     3361 agacgttttg cagcagcagt cgcttcacgt tcgctcgcgt atcggtgatt cattctgcta
     3421 accagtaagg caaccccgcc agcctagccg ggtcctcaac gacaggagca cgatcatgcg
     3481 cacccgtggc caggacccaa cgctgcccga gatgcgccgc gtgcggctgc tggagatggc
     3541 ggacgcgatg gatatgttct gccaagggtt ggtttgcgca ttcacagttc tccgcaagaa
     3601 ttgattggct ccaattcttg gagtggtgaa tccgttagcg aggtgccgcc ggcttccatt
     3661 caggtcgagg tggcccggct ccatgcaccg cgacgcaacg cggggaggca gacaaggtat
     3721 agggcggcgc ctacaatcca tgccaacccg ttccatgtgc tcgccgaggc ggcataaatc
     3781 gccgtgacga tcagcggtcc aatgatcgaa gttaggctgg taagagccgc gagcgatcct
     3841 tgaagctgtc cctgatggtc gtcatctacc tgcctggaca gcatggcctg caacgcgggc
     3901 atcccgatgc cgccggaagc gagaagaatc ataatgggga aggccatcca gcctcgcgtc
     3961 gcgaacgcca gcaagacgta gcccagcgcg tcggccgcca tgccggcgat aatggcctgc
     4021 ttctcgccga aacgtttggt ggcgggacca gtgacgaagg cttgagcgag ggcgtgcaag
     4081 attccgaata ccgcaagcga caggccgatc atcgtcgcgc tccagcgaaa gcggtcctcg
     4141 ccgaaaatga cccagagcgc tgccggcacc tgtcctacga gttgcatgat aaagaagaca
     4201 gtcataagtg cggcgacgat agtcatgccc cgcgcccacc ggaaggagct gactgggttg
     4261 aaggctctca agggcatcgg tcgagatccc ggtgcctaat gagtgagcta acttacatta
     4321 attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa
     4381 tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgccaggg tggtttttct
     4441 tttcaccagt gagacgggca acagctgatt gcccttcacc gcctggccct gagagagttg
     4501 cagcaagcgg tccacgctgg tttgccccag caggcgaaaa tcctgtttga tggtggttaa
     4561 cggcgggata taacatgagc tgtcttcggt atcgtcgtat cccactaccg agatatccgc
     4621 accaacgcgc agcccggact cggtaatggc gcgcattgcg cccagcgcca tctgatcgtt
     4681 ggcaaccagc atcgcagtgg gaacgatgcc ctcattcagc atttgcatgg tttgttgaaa
     4741 accggacatg gcactccagt cgccttcccg ttccgctatc ggctgaattt gattgcgagt
     4801 gagatattta tgccagccag ccagacgcag acgcgccgag acagaactta atgggcccgc
     4861 taacagcgcg atttgctggt gacccaatgc gaccagatgc tccacgccca gtcgcgtacc
     4921 gtcttcatgg gagaaaataa tactgttgat gggtgtctgg tcagagacat caagaaataa
     4981 cgccggaaca ttagtgcagg cagcttccac agcaatggca tcctggtcat ccagcggata
     5041 gttaatgatc agcccactga cgcgttgcgc gagaagattg tgcaccgccg ctttacaggc
     5101 ttcgacgccg cttcgttcta ccatcgacac caccacgctg gcacccagtt gatcggcgcg
     5161 agatttaatc gccgcgacaa tttgcgacgg cgcgtgcagg gccagactgg aggtggcaac
     5221 gccaatcagc aacgactgtt tgcccgccag ttgttgtgcc acgcggttgg gaatgtaatt
     5281 cagctccgcc atcgccgctt ccactttttc ccgcgttttc gcagaaacgt ggctggcctg
     5341 gttcaccacg cgggaaacgg tctgataaga gacaccggca tactctgcga catcgtataa
     5401 cgttactggt ttcacattca ccaccctgaa ttgactctct tccgggcgct atcatgccat
     5461 accgcgaaag gttttgcgcc attcgatggt gtccgggatc tcgacgctct cccttatgcg
     5521 actcctgcat taggaagcag cccagtagta ggttgaggcc gttgagcacc gccgccgcaa
     5581 ggaatggtgc atgcaaggag atggcgccca acagtccccc ggccacgggg cctgccacca
     5641 tacccacgcc gaaacaagcg ctcatgagcc cgaagtggcg agcccgatct tccccatcgg
     5701 tgatgtcggc gatataggcg ccagcaaccg cacctgtggc gccggtgatg ccggccacga
     5761 tgcgtccggc gtagaggatc gagatctcga tcccgcgaaa ttaatacgac tcactatagg
     5821 ggaattgtga gcggataaca attcccctct agaaataatt ttgtttaact ttaagaagga
     5881 gatatacata tggatgtatt catgaaagga ctttcaaagg ccaaggaggg agttgtggct
     5941 gctgctgaga aaaccaaaca gggtgtggca gaagcagcag gaaagacaaa agagggtgtt
     6001 ctctatgtag gctccaaaac caaggaggga gtggtgcatg gtgtggcaac agtggctgag
     6061 aagaccaaag agcaagtgac aaatgttgga ggagcagtgg tgacgggtgt gacagcagta
     6121 gcc
//
