LOCUS       sequence-207276-        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 Q109N (TAC-->TAT)
FEATURES             Location/Qualifiers
     source          1..6123
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      91..678
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     579..598
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     832..849
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    864..1003
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(929..948)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     1089..1111
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(1105..1296)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(2608..3690)
                     /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     3652..3671
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(3691..3768)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     3974..3993
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     4077..4096
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        4077..4095
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    4096..4120
                     /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             4135..4157
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             4591..4623
                     /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(4701..4719)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      4715..4762
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        complement(join(4937..4942,4943..4959,4960..4965))
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     primer_bind     complement(4992..5010)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5078..5182
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5183..5251,5252..6043)
                     /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(5401..5420)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc
       61 agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg
      121 ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct
      181 accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa atactgtcct
      241 tctagtgtag ccgtagttag gccaccactt caagaacctg tagcaccgcc tacatacctc
      301 gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg
      361 ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg
      421 tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag
      481 ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc
      541 agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat
      601 agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg
      661 gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc
      721 tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtatt
      781 accgcctttg agtgagctga taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca
      841 gtgagcgagg aagcggaaga gcgcctgatg cggtattttc tccttacgca tctgtgcggt
      901 atttcacacc gcaatggtgc actctcagta caatctgctc tgatgccgca tagttaagcc
      961 agtatacact ccgctatcgc tacgtgactg ggtcatggct gcgccccgac acccgccaac
     1021 acccgctgac gcgccctgac gggcttgtct gctcccggca tccgcttaca gacaagctgt
     1081 gaccgtctcc gggagctgca tgtgtcagag gttttcaccg tcatcaccga aacgcgcgag
     1141 gcagctgcgg taaagctcat cagcgtggtc gtgaagcgat tcacagatgt ctgcctgttc
     1201 atccgcgtcc agctcgttga gtttctccag aagcgttaat gtctggcttc tgataaagcg
     1261 ggccatgtta agggcggttt tttcctgttt ggtcactgat gcctccgtgt aagggggatt
     1321 tctgttcatg ggggtaatga taccgatgaa acgagagagg atgctcacga tacgggttac
     1381 tgatgatgaa catgcccggt tactggaacg ttgtgagggt aaacaactgg cggtatggat
     1441 gcggcgggac cagagaaaaa tcactcaggg tcaatgccag cgcttcgtta atacagatgt
     1501 aggtgttcca cagggtagcc agcagcatcc tgcgatgcag atccggaaca taatggtgca
     1561 gggcgctgac ttccgcgttt ccagacttta cgaaacacgg aaaccgaaga ccattcatgt
     1621 tgttgctcag gtcgcagacg ttttgcagca gcagtcgctt cacgttcgct cgcgtatcgg
     1681 tgattcattc tgctaaccag taaggcaacc ccgccagcct agccgggtcc tcaacgacag
     1741 gagcacgatc atgcgcaccc gtggccagga cccaacgctg cccgagatgc gccgcgtgcg
     1801 gctgctggag atggcggacg cgatggatat gttctgccaa gggttggttt gcgcattcac
     1861 agttctccgc aagaattgat tggctccaat tcttggagtg gtgaatccgt tagcgaggtg
     1921 ccgccggctt ccattcaggt cgaggtggcc cggctccatg caccgcgacg caacgcgggg
     1981 aggcagacaa ggtatagggc ggcgcctaca atccatgcca acccgttcca tgtgctcgcc
     2041 gaggcggcat aaatcgccgt gacgatcagc ggtccaatga tcgaagttag gctggtaaga
     2101 gccgcgagcg atccttgaag ctgtccctga tggtcgtcat ctacctgcct ggacagcatg
     2161 gcctgcaacg cgggcatccc gatgccgccg gaagcgagaa gaatcataat ggggaaggcc
     2221 atccagcctc gcgtcgcgaa cgccagcaag acgtagccca gcgcgtcggc cgccatgccg
     2281 gcgataatgg cctgcttctc gccgaaacgt ttggtggcgg gaccagtgac gaaggcttga
     2341 gcgagggcgt gcaagattcc gaataccgca agcgacaggc cgatcatcgt cgcgctccag
     2401 cgaaagcggt cctcgccgaa aatgacccag agcgctgccg gcacctgtcc tacgagttgc
     2461 atgataaaga agacagtcat aagtgcggcg acgatagtca tgccccgcgc ccaccggaag
     2521 gagctgactg ggttgaaggc tctcaagggc atcggtcgag atcccggtgc ctaatgagtg
     2581 agctaactta cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg
     2641 tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc
     2701 cagggtggtt tttcttttca ccagtgagac gggcaacagc tgattgccct tcaccgcctg
     2761 gccctgagag agttgcagca agcggtccac gctggtttgc cccagcaggc gaaaatcctg
     2821 tttgatggtg gttaacggcg ggatataaca tgagctgtct tcggtatcgt cgtatcccac
     2881 taccgagata tccgcaccaa cgcgcagccc ggactcggta atggcgcgca ttgcgcccag
     2941 cgccatctga tcgttggcaa ccagcatcgc agtgggaacg atgccctcat tcagcatttg
     3001 catggtttgt tgaaaaccgg acatggcact ccagtcgcct tcccgttccg ctatcggctg
     3061 aatttgattg cgagtgagat atttatgcca gccagccaga cgcagacgcg ccgagacaga
     3121 acttaatggg cccgctaaca gcgcgatttg ctggtgaccc aatgcgacca gatgctccac
     3181 gcccagtcgc gtaccgtctt catgggagaa aataatactg ttgatgggtg tctggtcaga
     3241 gacatcaaga aataacgccg gaacattagt gcaggcagct tccacagcaa tggcatcctg
     3301 gtcatccagc ggatagttaa tgatcagccc actgacgcgt tgcgcgagaa gattgtgcac
     3361 cgccgcttta caggcttcga cgccgcttcg ttctaccatc gacaccacca cgctggcacc
     3421 cagttgatcg gcgcgagatt taatcgccgc gacaatttgc gacggcgcgt gcagggccag
     3481 actggaggtg gcaacgccaa tcagcaacga ctgtttgccc gccagttgtt gtgccacgcg
     3541 gttgggaatg taattcagct ccgccatcgc cgcttccact ttttcccgcg ttttcgcaga
     3601 aacgtggctg gcctggttca ccacgcggga aacggtctga taagagacac cggcatactc
     3661 tgcgacatcg tataacgtta ctggtttcac attcaccacc ctgaattgac tctcttccgg
     3721 gcgctatcat gccataccgc gaaaggtttt gcgccattcg atggtgtccg ggatctcgac
     3781 gctctccctt atgcgactcc tgcattagga agcagcccag tagtaggttg aggccgttga
     3841 gcaccgccgc cgcaaggaat ggtgcatgca aggagatggc gcccaacagt cccccggcca
     3901 cggggcctgc caccataccc acgccgaaac aagcgctcat gagcccgaag tggcgagccc
     3961 gatcttcccc atcggtgatg tcggcgatat aggcgccagc aaccgcacct gtggcgccgg
     4021 tgatgccggc cacgatgcgt ccggcgtaga ggatcgagat ctcgatcccg cgaaattaat
     4081 acgactcact ataggggaat tgtgagcgga taacaattcc cctctagaaa taattttgtt
     4141 taactttaag aaggagatat acatatggat gtattcatga aaggactttc aaaggccaag
     4201 gagggagttg tggctgctgc tgagaaaacc aaacagggtg tggcagaagc agcaggaaag
     4261 acaaaagagg gtgttctcta tgtaggctcc aaaaccaagg agggagtggt gcatggtgtg
     4321 gcaacagtgg ctgagaagac caaagagcaa gtgacaaatg ttggaggagc agtggtgacg
     4381 ggtgtgacag cagtagccca gaagacagtg gagggagcgg ggagcattgc agcagccact
     4441 ggctttgtca aaaaggacca gttgggcaag aatgaagaag gagccccaaa cgaaggaatt
     4501 ctggaagata tgcctgtgga tcctgacaat gaggcttatg aaatgccttc tgaggaaggg
     4561 tatcaagact atgaacctga agcctaacat atggctagca tgactggtgg acagcaaatg
     4621 ggtcgcggat cccgggtacc tcgagtcgac cgatccggct gctaacaaag cccgaaagga
     4681 agctgagttg gctgctgcca ccgctgagca ataactagca taaccccttg gggcctctaa
     4741 acgggtcttg aggggttttt tgctgaaagg aggaactata tccggatatc ccgcaagagg
     4801 cccggcagta ccggcataac caagcctatg cctacagcat ccagggtgac ggtgccgagg
     4861 atgacgatga gcgcattgtt agatttcata cacggtgcct gactgcgtta gcaatttaac
     4921 tgtgataaac taccgcatta aagcttatcg atgataagct gtcaaacatg agaattcttg
     4981 aagacgaaag ggcctcgtga tacgcctatt tttataggtt aatgtcatga taataatggt
     5041 ttcttagacg tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt
     5101 tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca
     5161 ataatattga aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc ttattccctt
     5221 ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctggtga aagtaaaaga
     5281 tgctgaagat cagttgggtg cacgagtggg ttacatcgaa ctggatctca acagcggtaa
     5341 gatccttgag agttttcgcc ccgaagaacg ttttccaatg atgagcactt ttaaagttct
     5401 gctatgtggc gcggtattat cccgtgttga cgccgggcaa gagcaactcg gtcgccgcat
     5461 acactattct cagaatgact tggttgagta ctcaccagtc acagaaaagc atcttacgga
     5521 tggcatgaca gtaagagaat tatgcagtgc tgccataacc atgagtgata acactgcggc
     5581 caacttactt ctgacaacga tcggaggacc gaaggagcta accgcttttt tgcacaacat
     5641 gggggatcat gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa
     5701 cgacgagcgt gacaccacga tgcctgcagc aatggcaaca acgttgcgca aactattaac
     5761 tggcgaacta cttactctag cttcccggca acaattaata gactggatgg aggcggataa
     5821 agttgcagga ccacttctgc gctcggccct tccggctggc tggtttattg ctgataaatc
     5881 tggagccggt gagcgtgggt ctcgcggtat cattgcagca ctggggccag atggtaagcc
     5941 ctcccgtatc gtagttatct acacgacggg gagtcaggca actatggatg aacgaaatag
     6001 acagatcgct gagataggtg cctcactgat taagcattgg taactgtcag accaagttta
     6061 ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa
     6121 gat
//
