LOCUS       Exported                3651 bp ds-DNA     circular SYN 13-MAY-2021
DEFINITION  Bacterial expression of mutant human alpha synuclein.
ACCESSION   .
VERSION     .
KEYWORDS    pT7-7 asyn WT1-106 Mxe
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3651)
  TITLE     Lashuel Lab plasmids
REFERENCE   2  (bases 1 to 3651)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 13, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..3651
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     rep_origin      10..598
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     499..518
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     752..769
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     1004..1026
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     1091..1110
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1091..1109
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             1141..1163
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             1489..2082
                     /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             2113..2268
                     /codon_start=1
                     /gene="Bacillus circulans chiA"
                     /product="chitin binding domain from chitinase A1 (Watanabe
                     et al., 1994)"
                     /label=CBD
                     /translation="TTNPGVSAWQVNTAYTAGQLVTYNGKTYKCLQPHTSLAGWEPSNV
                     PALWQLQ"
     CDS             complement(2481..3341)
                     /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     3104..3123
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3342..3446)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3514..3532
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc
       61 accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt
      121 aactggcttc agcagagcgc agataccaaa tactgtcctt ctagtgtagc cgtagttagg
      181 ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc
      241 agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt
      301 accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga
      361 gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct
      421 tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg
      481 cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca
      541 cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa
      601 cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt
      661 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga
      721 taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga
      781 gcgcctgatg cggtattttc tccttacgca tctgtgcggt atttcacacc gcataggaag
      841 atcttccgga agatcttcct atggtgcact ctcagtacaa tctgctctga tgcgctacgt
      901 gactgggtca tggctgcgcc ccgacacccg ccaacacccg ctgacgcgcc ctgacgggct
      961 tgtctgctcc cggcatccgc ttacagacaa gctgtgaccg tctccgggag ctgcatgtgt
     1021 cagaggtttt caccgtcatc accgaaacgc gcgaggccca gcgattcgaa cttctgatag
     1081 acttcgaaat taatacgact cactataggg agaccacaac ggtttccctc tagaaataat
     1141 tttgtttaac tttaagaagg agatatacat atggatgtat tcatgaaagg actttcaaag
     1201 gccaaggagg gagttgtggc tgctgctgag aaaaccaaac agggtgtggc agaagcagca
     1261 ggaaagacaa aagagggtgt tctctatgta ggctccaaaa ccaaggaggg agtggtgcat
     1321 ggtgtggcaa cagtggctga gaagaccaaa gagcaagtga caaatgttgg aggagcagtg
     1381 gtgacgggtg tgacagcagt agcccagaag acagtggagg gagcagggag cattgcagca
     1441 gccactggct ttgtcaaaaa ggaccagttg ggcaagaatg aagaaggatg catcacggga
     1501 gatgcactag ttgccctacc cgagggcgag tcggtacgca tcgccgacat cgtgccgggt
     1561 gcgcggccca acagtgacaa cgccatcgac ctgaaagtcc ttgaccggca tggcaatccc
     1621 gtgctcgccg accggctgtt ccactccggc gagcatccgg tgtacacggt gcgtacggtc
     1681 gaaggtctgc gtgtgacggg caccgcgaac cacccgttgt tgtgtttggt cgacgtcgcc
     1741 ggggtgccga ccctgctgtg gaagctgatc gacgaaatca agccgggcga ttacgcggtg
     1801 attcaacgca gcgcattcag cgtcgactgt gcaggttttg cccgcgggaa acccgaattt
     1861 gcgcccacaa cctacacagt cggcgtccct ggactggtgc gtttcttgga agcacaccac
     1921 cgagacccgg acgcccaagc tatcgccgac gagctgaccg acgggcggtt ctactacgcg
     1981 aaagtcgcca gtgtcaccga cgccggcgtg cagccggtgt atagccttcg tgtcgacacg
     2041 gcagaccacg cgtttatcac gaacgggttc gtcagccacg ctactggcct caccggtctg
     2101 tagtgaggcc tcacgacaaa tcctggtgta tccgcttggc aggtcaacac agcttatact
     2161 gcgggacaat tggtcacata taacggcaag acgtataaat gtttgcagcc ccacacctcc
     2221 ttggcaggat gggaaccatc caacgttcct gccttgtggc agcttcaatg actgcaggaa
     2281 ggggatcctg acaatgaggc ttatgaaatg ccttctgagg aagggtatca agactacgaa
     2341 cctgaagcct aagaaatatc tttgctccca gtttcttgag atctgctgac agatgttcca
     2401 tccaagctta tcgatgataa gctgtcaaac atgagaatta aatcaatcta aagtatatat
     2461 gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc
     2521 tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg
     2581 gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct
     2641 ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca
     2701 actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg
     2761 ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg
     2821 tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc
     2881 cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag
     2941 ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg
     3001 ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag
     3061 tgtatgcggc gaccgagttg ctcttgcccg gcgtcaacac gggataatac cgcgccacat
     3121 agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg
     3181 atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca
     3241 gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca
     3301 aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat
     3361 tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag
     3421 aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgtctaa
     3481 gaaaccatta ttatcatgac attaacctat aaaaataggc gtatcacgag gccctttcgt
     3541 cttcaagaat aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc
     3601 ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca a
//
