LOCUS       Exported                6123 bp ds-DNA     circular SYN 13-MAY-2021
DEFINITION  Bacterial expression of mutant human alpha synuclein.
ACCESSION   .
VERSION     .
KEYWORDS    pET11a asyn Q109E (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      28..615
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     516..535
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     769..786
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    801..940
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(866..885)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     1026..1048
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(1042..1233)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(2545..3627)
                     /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     3589..3608
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(3628..3705)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     3911..3930
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     4014..4033
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        4014..4032
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    4033..4057
                     /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             4072..4094
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             4528..4560
                     /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(4638..4656)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      4652..4699
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        complement(4874..4902)
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     primer_bind     complement(4929..4947)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5015..5119
                     /gene="bla"
                     /label=AmpR promoter
     CDS             5120..5980
                     /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(5338..5357)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg
       61 cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc
      121 aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgtccttct
      181 agtgtagccg tagttaggcc accacttcaa gaacctgtag caccgcctac atacctcgct
      241 ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg
      301 gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc
      361 acacagccca gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta
      421 tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg
      481 gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt
      541 cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg
      601 cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg
      661 ccttttgctc acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc
      721 gcctttgagt gagctgatac cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg
      781 agcgaggaag cggaagagcg cctgatgcgg tattttctcc ttacgcatct gtgcggtatt
      841 tcacaccgca atggtgcact ctcagtacaa tctgctctga tgccgcatag ttaagccagt
      901 atacactccg ctatcgctac gtgactgggt catggctgcg ccccgacacc cgccaacacc
      961 cgctgacgcg ccctgacggg cttgtctgct cccggcatcc gcttacagac aagctgtgac
     1021 cgtctccggg agctgcatgt gtcagaggtt ttcaccgtca tcaccgaaac gcgcgaggca
     1081 gctgcggtaa agctcatcag cgtggtcgtg aagcgattca cagatgtctg cctgttcatc
     1141 cgcgtccagc tcgttgagtt tctccagaag cgttaatgtc tggcttctga taaagcgggc
     1201 catgttaagg gcggtttttt cctgtttggt cactgatgcc tccgtgtaag ggggatttct
     1261 gttcatgggg gtaatgatac cgatgaaacg agagaggatg ctcacgatac gggttactga
     1321 tgatgaacat gcccggttac tggaacgttg tgagggtaaa caactggcgg tatggatgcg
     1381 gcgggaccag agaaaaatca ctcagggtca atgccagcgc ttcgttaata cagatgtagg
     1441 tgttccacag ggtagccagc agcatcctgc gatgcagatc cggaacataa tggtgcaggg
     1501 cgctgacttc cgcgtttcca gactttacga aacacggaaa ccgaagacca ttcatgttgt
     1561 tgctcaggtc gcagacgttt tgcagcagca gtcgcttcac gttcgctcgc gtatcggtga
     1621 ttcattctgc taaccagtaa ggcaaccccg ccagcctagc cgggtcctca acgacaggag
     1681 cacgatcatg cgcacccgtg gccaggaccc aacgctgccc gagatgcgcc gcgtgcggct
     1741 gctggagatg gcggacgcga tggatatgtt ctgccaaggg ttggtttgcg cattcacagt
     1801 tctccgcaag aattgattgg ctccaattct tggagtggtg aatccgttag cgaggtgccg
     1861 ccggcttcca ttcaggtcga ggtggcccgg ctccatgcac cgcgacgcaa cgcggggagg
     1921 cagacaaggt atagggcggc gcctacaatc catgccaacc cgttccatgt gctcgccgag
     1981 gcggcataaa tcgccgtgac gatcagcggt ccaatgatcg aagttaggct ggtaagagcc
     2041 gcgagcgatc cttgaagctg tccctgatgg tcgtcatcta cctgcctgga cagcatggcc
     2101 tgcaacgcgg gcatcccgat gccgccggaa gcgagaagaa tcataatggg gaaggccatc
     2161 cagcctcgcg tcgcgaacgc cagcaagacg tagcccagcg cgtcggccgc catgccggcg
     2221 ataatggcct gcttctcgcc gaaacgtttg gtggcgggac cagtgacgaa ggcttgagcg
     2281 agggcgtgca agattccgaa taccgcaagc gacaggccga tcatcgtcgc gctccagcga
     2341 aagcggtcct cgccgaaaat gacccagagc gctgccggca cctgtcctac gagttgcatg
     2401 ataaagaaga cagtcataag tgcggcgacg atagtcatgc cccgcgccca ccggaaggag
     2461 ctgactgggt tgaaggctct caagggcatc ggtcgagatc ccggtgccta atgagtgagc
     2521 taacttacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc
     2581 cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgccag
     2641 ggtggttttt cttttcacca gtgagacggg caacagctga ttgcccttca ccgcctggcc
     2701 ctgagagagt tgcagcaagc ggtccacgct ggtttgcccc agcaggcgaa aatcctgttt
     2761 gatggtggtt aacggcggga tataacatga gctgtcttcg gtatcgtcgt atcccactac
     2821 cgagatatcc gcaccaacgc gcagcccgga ctcggtaatg gcgcgcattg cgcccagcgc
     2881 catctgatcg ttggcaacca gcatcgcagt gggaacgatg ccctcattca gcatttgcat
     2941 ggtttgttga aaaccggaca tggcactcca gtcgccttcc cgttccgcta tcggctgaat
     3001 ttgattgcga gtgagatatt tatgccagcc agccagacgc agacgcgccg agacagaact
     3061 taatgggccc gctaacagcg cgatttgctg gtgacccaat gcgaccagat gctccacgcc
     3121 cagtcgcgta ccgtcttcat gggagaaaat aatactgttg atgggtgtct ggtcagagac
     3181 atcaagaaat aacgccggaa cattagtgca ggcagcttcc acagcaatgg catcctggtc
     3241 atccagcgga tagttaatga tcagcccact gacgcgttgc gcgagaagat tgtgcaccgc
     3301 cgctttacag gcttcgacgc cgcttcgttc taccatcgac accaccacgc tggcacccag
     3361 ttgatcggcg cgagatttaa tcgccgcgac aatttgcgac ggcgcgtgca gggccagact
     3421 ggaggtggca acgccaatca gcaacgactg tttgcccgcc agttgttgtg ccacgcggtt
     3481 gggaatgtaa ttcagctccg ccatcgccgc ttccactttt tcccgcgttt tcgcagaaac
     3541 gtggctggcc tggttcacca cgcgggaaac ggtctgataa gagacaccgg catactctgc
     3601 gacatcgtat aacgttactg gtttcacatt caccaccctg aattgactct cttccgggcg
     3661 ctatcatgcc ataccgcgaa aggttttgcg ccattcgatg gtgtccggga tctcgacgct
     3721 ctcccttatg cgactcctgc attaggaagc agcccagtag taggttgagg ccgttgagca
     3781 ccgccgccgc aaggaatggt gcatgcaagg agatggcgcc caacagtccc ccggccacgg
     3841 ggcctgccac catacccacg ccgaaacaag cgctcatgag cccgaagtgg cgagcccgat
     3901 cttccccatc ggtgatgtcg gcgatatagg cgccagcaac cgcacctgtg gcgccggtga
     3961 tgccggccac gatgcgtccg gcgtagagga tcgagatctc gatcccgcga aattaatacg
     4021 actcactata ggggaattgt gagcggataa caattcccct ctagaaataa ttttgtttaa
     4081 ctttaagaag gagatataca tatggatgta ttcatgaaag gactttcaaa ggccaaggag
     4141 ggagttgtgg ctgctgctga gaaaaccaaa cagggtgtgg cagaagcagc aggaaagaca
     4201 aaagagggtg ttctctatgt aggctccaaa accaaggagg gagtggtgca tggtgtggca
     4261 acagtggctg agaagaccaa agagcaagtg acaaatgttg gaggagcagt ggtgacgggt
     4321 gtgacagcag tagcccagaa gacagtggag ggagcgggga gcattgcagc agccactggc
     4381 tttgtcaaaa aggaccagtt gggcaagaat gaagaaggag ccccagagga aggaattctg
     4441 gaagatatgc ctgtggatcc tgacaatgag gcttatgaaa tgccttctga ggaagggtat
     4501 caagactatg aacctgaagc ctaacatatg gctagcatga ctggtggaca gcaaatgggt
     4561 cgcggatccc gggtacctcg agtcgaccga tccggctgct aacaaagccc gaaaggaagc
     4621 tgagttggct gctgccaccg ctgagcaata actagcataa ccccttgggg cctctaaacg
     4681 ggtcttgagg ggttttttgc tgaaaggagg aactatatcc ggatatcccg caagaggccc
     4741 ggcagtaccg gcataaccaa gcctatgcct acagcatcca gggtgacggt gccgaggatg
     4801 acgatgagcg cattgttaga tttcatacac ggtgcctgac tgcgttagca atttaactgt
     4861 gataaactac cgcattaaag cttatcgatg ataagctgtc aaacatgaga attcttgaag
     4921 acgaaagggc ctcgtgatac gcctattttt ataggttaat gtcatgataa taatggtttc
     4981 ttagacgtca ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt
     5041 ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata
     5101 atattgaaaa aggaagagta tgagtattca acatttccgt gtcgccctta ttcccttttt
     5161 tgcggcattt tgccttcctg tttttgctca cccagaaacg ctggtgaaag taaaagatgc
     5221 tgaagatcag ttgggtgcac gagtgggtta catcgaactg gatctcaaca gcggtaagat
     5281 ccttgagagt tttcgccccg aagaacgttt tccaatgatg agcactttta aagttctgct
     5341 atgtggcgcg gtattatccc gtgttgacgc cgggcaagag caactcggtc gccgcataca
     5401 ctattctcag aatgacttgg ttgagtactc accagtcaca gaaaagcatc ttacggatgg
     5461 catgacagta agagaattat gcagtgctgc cataaccatg agtgataaca ctgcggccaa
     5521 cttacttctg acaacgatcg gaggaccgaa ggagctaacc gcttttttgc acaacatggg
     5581 ggatcatgta actcgccttg atcgttggga accggagctg aatgaagcca taccaaacga
     5641 cgagcgtgac accacgatgc ctgcagcaat ggcaacaacg ttgcgcaaac tattaactgg
     5701 cgaactactt actctagctt cccggcaaca attaatagac tggatggagg cggataaagt
     5761 tgcaggacca cttctgcgct cggcccttcc ggctggctgg tttattgctg ataaatctgg
     5821 agccggtgag cgtgggtctc gcggtatcat tgcagcactg gggccagatg gtaagccctc
     5881 ccgtatcgta gttatctaca cgacggggag tcaggcaact atggatgaac gaaatagaca
     5941 gatcgctgag ataggtgcct cactgattaa gcattggtaa ctgtcagacc aagtttactc
     6001 atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat
     6061 cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc
     6121 aga
//
