LOCUS       Exported                3499 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  Bacterial expression of mutant human alpha synuclein.
ACCESSION   .
VERSION     .
KEYWORDS    pT7-7 asyn 1-106AA Mxe Histag
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3499)
  TITLE     Lashuel Lab plasmids
REFERENCE   2  (bases 1 to 3499)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..3499
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     rep_origin      35..623
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     524..543
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     777..794
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     1029..1051
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     1116..1135
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1116..1134
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             1166..1188
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             1514..2107
                     /codon_start=1
                     /gene="Mycobacterium xenopi gyrA"
                     /product="modified GyrA intein (Southworth et al., 1999)"
                     /label=Mxe GyrA intein
                     /note="N-terminal cleavage of this intein can be induced by
                     thiols."
                     /translation="CITGDALVALPEGESVRIADIVPGARPNSDNAIDLKVLDRHGNPV
                     LADRLFHSGEHPVYTVRTVEGLRVTGTANHPLLCLVDVAGVPTLLWKLIDEIKPGDYAV
                     IQRSAFSVDCAGFARGKPEFAPTTYTVGVPGLVRFLEAHHRDPDAQAIADELTDGRFYY
                     AKVASVTDAGVQPVYSLRVDTADHAFITNGFVSHA"
     CDS             2132..2149
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             complement(2354..3214)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     2977..2996
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3215..3319)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3387..3405
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa
       61 tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag
      121 agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg
      181 tccttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat
      241 acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta
      301 ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg
      361 gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc
      421 gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa
      481 gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc
      541 tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt
      601 caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct
      661 tttgctggcc ttttgctcac atgttctttc ctgcgttatc ccctgattct gtggataacc
      721 gtattaccgc ctttgagtga gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg
      781 agtcagtgag cgaggaagcg gaagagcgcc tgatgcggta ttttctcctt acgcatctgt
      841 gcggtatttc acaccgcata ggaagatctt ccggaagatc ttcctatggt gcactctcag
      901 tacaatctgc tctgatgcgc tacgtgactg ggtcatggct gcgccccgac acccgccaac
      961 acccgctgac gcgccctgac gggcttgtct gctcccggca tccgcttaca gacaagctgt
     1021 gaccgtctcc gggagctgca tgtgtcagag gttttcaccg tcatcaccga aacgcgcgag
     1081 gcccagcgat tcgaacttct gatagacttc gaaattaata cgactcacta tagggagacc
     1141 acaacggttt ccctctagaa ataattttgt ttaactttaa gaaggagata tacatatgga
     1201 tgtattcatg aaaggacttt caaaggccaa ggagggagtt gtggctgctg ctgagaaaac
     1261 caaacagggt gtggcagaag cagcaggaaa gacaaaagag ggtgttctct atgtaggctc
     1321 caaaaccaag gagggagtgg tgcatggtgt ggcaacagtg gctgagaaga ccaaagagca
     1381 agtgacaaat gttggaggag cagtggtgac gggtgtgaca gcagtagccc agaagacagt
     1441 ggagggagca gggagcattg cagcagccac tggctttgtc aaaaaggacc agttgggcaa
     1501 gaatgaagaa ggatgcatca cgggagatgc actagttgcc ctacccgagg gcgagtcggt
     1561 acgcatcgcc gacatcgtgc cgggtgcgcg gcccaacagt gacaacgcca tcgacctgaa
     1621 agtccttgac cggcatggca atcccgtgct cgccgaccgg ctgttccact ccggcgagca
     1681 tccggtgtac acggtgcgta cggtcgaagg tctgcgtgtg acgggcaccg cgaaccaccc
     1741 gttgttgtgt ttggtcgacg tcgccggggt gccgaccctg ctgtggaagc tgatcgacga
     1801 aatcaagccg ggcgattacg cggtgattca acgcagcgca ttcagcgtcg actgtgcagg
     1861 ttttgcccgc gggaaacccg aatttgcgcc cacaacctac acagtcggcg tccctggact
     1921 ggtgcgtttc ttggaagcac accaccgaga cccggacgcc caagctatcg ccgacgagct
     1981 gaccgacggg cggttctact acgcgaaagt cgccagtgtc accgacgccg gcgtgcagcc
     2041 ggtgtatagc cttcgtgtcg acacggcaga ccacgcgttt atcacgaacg ggttcgtcag
     2101 ccacgctact ggcctcaccg gtctggaatt ccaccaccac caccaccact gataaggatc
     2161 ctgacaatga ggcttatgaa atgccttctg aggaagggta tcaagactac gaacctgaag
     2221 cctaagaaat atctttgctc ccagtttctt gagatctgct gacagatgtt ccatccaagc
     2281 ttatcgatga taagctgtca aacatgagaa ttaaatcaat ctaaagtata tatgagtaaa
     2341 cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat
     2401 ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct
     2461 taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt
     2521 tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat
     2581 ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta
     2641 atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg
     2701 gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt
     2761 tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg
     2821 cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg
     2881 taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc
     2941 ggcgaccgag ttgctcttgc ccggcgtcaa cacgggataa taccgcgcca catagcagaa
     3001 ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac
     3061 cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt
     3121 ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg
     3181 gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa
     3241 gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata
     3301 aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc taagaaacca
     3361 ttattatcat gacattaacc tataaaaata ggcgtatcac gaggcccttt cgtcttcaag
     3421 aataaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt
     3481 gagttttcgt tccactgag
//
