LOCUS       Exported                6123 bp ds-DNA     circular SYN 13-MAY-2021
DEFINITION  Bacterial expression of mutant human alpha synuclein.
ACCESSION   .
VERSION     .
KEYWORDS    pET11a asyn Q79N (TAC-->TAT)
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 May 13, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6123
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     rep_origin      62..649
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     550..569
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     803..820
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    835..974
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(900..919)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     1060..1082
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(1076..1267)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(2579..3661)
                     /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     3623..3642
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(3662..3739)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     3945..3964
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     4048..4067
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        4048..4066
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    4067..4091
                     /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             4106..4128
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             4562..4594
                     /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(4672..4690)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      4686..4733
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        complement(4908..4936)
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     primer_bind     complement(4963..4981)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5049..5153
                     /gene="bla"
                     /label=AmpR promoter
     CDS             5154..6014
                     /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(5372..5391)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt
       61 cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac
      121 cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct
      181 tcagcagagc gcagatacca aatactgtcc ttctagtgta gccgtagtta ggccaccact
      241 tcaagaacct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
      301 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
      361 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
      421 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
      481 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
      541 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
      601 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
      661 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc
      721 gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg
      781 ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcctgat
      841 gcggtatttt ctccttacgc atctgtgcgg tatttcacac cgcaatggtg cactctcagt
      901 acaatctgct ctgatgccgc atagttaagc cagtatacac tccgctatcg ctacgtgact
      961 gggtcatggc tgcgccccga cacccgccaa cacccgctga cgcgccctga cgggcttgtc
     1021 tgctcccggc atccgcttac agacaagctg tgaccgtctc cgggagctgc atgtgtcaga
     1081 ggttttcacc gtcatcaccg aaacgcgcga ggcagctgcg gtaaagctca tcagcgtggt
     1141 cgtgaagcga ttcacagatg tctgcctgtt catccgcgtc cagctcgttg agtttctcca
     1201 gaagcgttaa tgtctggctt ctgataaagc gggccatgtt aagggcggtt ttttcctgtt
     1261 tggtcactga tgcctccgtg taagggggat ttctgttcat gggggtaatg ataccgatga
     1321 aacgagagag gatgctcacg atacgggtta ctgatgatga acatgcccgg ttactggaac
     1381 gttgtgaggg taaacaactg gcggtatgga tgcggcggga ccagagaaaa atcactcagg
     1441 gtcaatgcca gcgcttcgtt aatacagatg taggtgttcc acagggtagc cagcagcatc
     1501 ctgcgatgca gatccggaac ataatggtgc agggcgctga cttccgcgtt tccagacttt
     1561 acgaaacacg gaaaccgaag accattcatg ttgttgctca ggtcgcagac gttttgcagc
     1621 agcagtcgct tcacgttcgc tcgcgtatcg gtgattcatt ctgctaacca gtaaggcaac
     1681 cccgccagcc tagccgggtc ctcaacgaca ggagcacgat catgcgcacc cgtggccagg
     1741 acccaacgct gcccgagatg cgccgcgtgc ggctgctgga gatggcggac gcgatggata
     1801 tgttctgcca agggttggtt tgcgcattca cagttctccg caagaattga ttggctccaa
     1861 ttcttggagt ggtgaatccg ttagcgaggt gccgccggct tccattcagg tcgaggtggc
     1921 ccggctccat gcaccgcgac gcaacgcggg gaggcagaca aggtataggg cggcgcctac
     1981 aatccatgcc aacccgttcc atgtgctcgc cgaggcggca taaatcgccg tgacgatcag
     2041 cggtccaatg atcgaagtta ggctggtaag agccgcgagc gatccttgaa gctgtccctg
     2101 atggtcgtca tctacctgcc tggacagcat ggcctgcaac gcgggcatcc cgatgccgcc
     2161 ggaagcgaga agaatcataa tggggaaggc catccagcct cgcgtcgcga acgccagcaa
     2221 gacgtagccc agcgcgtcgg ccgccatgcc ggcgataatg gcctgcttct cgccgaaacg
     2281 tttggtggcg ggaccagtga cgaaggcttg agcgagggcg tgcaagattc cgaataccgc
     2341 aagcgacagg ccgatcatcg tcgcgctcca gcgaaagcgg tcctcgccga aaatgaccca
     2401 gagcgctgcc ggcacctgtc ctacgagttg catgataaag aagacagtca taagtgcggc
     2461 gacgatagtc atgccccgcg cccaccggaa ggagctgact gggttgaagg ctctcaaggg
     2521 catcggtcga gatcccggtg cctaatgagt gagctaactt acattaattg cgttgcgctc
     2581 actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg
     2641 cgcggggaga ggcggtttgc gtattgggcg ccagggtggt ttttcttttc accagtgaga
     2701 cgggcaacag ctgattgccc ttcaccgcct ggccctgaga gagttgcagc aagcggtcca
     2761 cgctggtttg ccccagcagg cgaaaatcct gtttgatggt ggttaacggc gggatataac
     2821 atgagctgtc ttcggtatcg tcgtatccca ctaccgagat atccgcacca acgcgcagcc
     2881 cggactcggt aatggcgcgc attgcgccca gcgccatctg atcgttggca accagcatcg
     2941 cagtgggaac gatgccctca ttcagcattt gcatggtttg ttgaaaaccg gacatggcac
     3001 tccagtcgcc ttcccgttcc gctatcggct gaatttgatt gcgagtgaga tatttatgcc
     3061 agccagccag acgcagacgc gccgagacag aacttaatgg gcccgctaac agcgcgattt
     3121 gctggtgacc caatgcgacc agatgctcca cgcccagtcg cgtaccgtct tcatgggaga
     3181 aaataatact gttgatgggt gtctggtcag agacatcaag aaataacgcc ggaacattag
     3241 tgcaggcagc ttccacagca atggcatcct ggtcatccag cggatagtta atgatcagcc
     3301 cactgacgcg ttgcgcgaga agattgtgca ccgccgcttt acaggcttcg acgccgcttc
     3361 gttctaccat cgacaccacc acgctggcac ccagttgatc ggcgcgagat ttaatcgccg
     3421 cgacaatttg cgacggcgcg tgcagggcca gactggaggt ggcaacgcca atcagcaacg
     3481 actgtttgcc cgccagttgt tgtgccacgc ggttgggaat gtaattcagc tccgccatcg
     3541 ccgcttccac tttttcccgc gttttcgcag aaacgtggct ggcctggttc accacgcggg
     3601 aaacggtctg ataagagaca ccggcatact ctgcgacatc gtataacgtt actggtttca
     3661 cattcaccac cctgaattga ctctcttccg ggcgctatca tgccataccg cgaaaggttt
     3721 tgcgccattc gatggtgtcc gggatctcga cgctctccct tatgcgactc ctgcattagg
     3781 aagcagccca gtagtaggtt gaggccgttg agcaccgccg ccgcaaggaa tggtgcatgc
     3841 aaggagatgg cgcccaacag tcccccggcc acggggcctg ccaccatacc cacgccgaaa
     3901 caagcgctca tgagcccgaa gtggcgagcc cgatcttccc catcggtgat gtcggcgata
     3961 taggcgccag caaccgcacc tgtggcgccg gtgatgccgg ccacgatgcg tccggcgtag
     4021 aggatcgaga tctcgatccc gcgaaattaa tacgactcac tataggggaa ttgtgagcgg
     4081 ataacaattc ccctctagaa ataattttgt ttaactttaa gaaggagata tacatatgga
     4141 tgtattcatg aaaggacttt caaaggccaa ggagggagtt gtggctgctg ctgagaaaac
     4201 caaacagggt gtggcagaag cagcaggaaa gacaaaagag ggtgttctct atgtaggctc
     4261 caaaaccaag gagggagtgg tgcatggtgt ggcaacagtg gctgagaaga ccaaagagca
     4321 agtgacaaat gttggaggag cagtggtgac gggtgtgaca gcagtagcca acaagacagt
     4381 ggagggagcg gggagcattg cagcagccac tggctttgtc aaaaaggacc agttgggcaa
     4441 gaatgaagaa ggagccccac aggaaggaat tctggaagat atgcctgtgg atcctgacaa
     4501 tgaggcttat gaaatgcctt ctgaggaagg gtatcaagac tatgaacctg aagcctaaca
     4561 tatggctagc atgactggtg gacagcaaat gggtcgcgga tcccgggtac ctcgagtcga
     4621 ccgatccggc tgctaacaaa gcccgaaagg aagctgagtt ggctgctgcc accgctgagc
     4681 aataactagc ataacccctt ggggcctcta aacgggtctt gaggggtttt ttgctgaaag
     4741 gaggaactat atccggatat cccgcaagag gcccggcagt accggcataa ccaagcctat
     4801 gcctacagca tccagggtga cggtgccgag gatgacgatg agcgcattgt tagatttcat
     4861 acacggtgcc tgactgcgtt agcaatttaa ctgtgataaa ctaccgcatt aaagcttatc
     4921 gatgataagc tgtcaaacat gagaattctt gaagacgaaa gggcctcgtg atacgcctat
     4981 ttttataggt taatgtcatg ataataatgg tttcttagac gtcaggtggc acttttcggg
     5041 gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc
     5101 tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag agtatgagta
     5161 ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg
     5221 ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg
     5281 gttacatcga actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac
     5341 gttttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtgttg
     5401 acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt
     5461 actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg
     5521 ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg atcggaggac
     5581 cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt
     5641 gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgcag
     5701 caatggcaac aacgttgcgc aaactattaa ctggcgaact acttactcta gcttcccggc
     5761 aacaattaat agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc
     5821 ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggg tctcgcggta
     5881 tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg
     5941 ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga
     6001 ttaagcattg gtaactgtca gaccaagttt actcatatat actttagatt gatttaaaac
     6061 ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa
     6121 tcc
//
