LOCUS       sequence-207309-        5552 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of human alpha synuclein.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5552)
  AUTHORS   Mbefo MK, Fares MB, Paleologou K, Oueslati A, Yin G, Tenreiro S, 
            Pinto M, Outeiro T, Zweckstetter M, Masliah E, Lashuel HA
  TITLE     Parkinson disease mutant E46K enhances alpha-synuclein 
            phosphorylation in mammalian cell lines, in yeast, and in vivo.
  JOURNAL   J Biol Chem. 2015 Apr 10;290(15):9412-27. doi: 
            10.1074/jbc.M114.610774. Epub 2015 Feb 5.
  PUBMED    25657004
REFERENCE   2  (bases 1 to 5552)
  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"; doi: 
            "10.1074/jbc.M114.610774"; journalName: "J Biol Chem"; date: 
            "2015-04-10- 10"; volume: "290"; issue: "15"; pages: "9412-27"
COMMENT     Sequence Label: pcDNA6 asyn WT
FEATURES             Location/Qualifiers
     source          1..5552
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        24..403
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        404..607
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     558..578
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     652..671
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        652..670
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             696..718
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     protein_bind    complement(1226..1250)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(1297..1314)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1303..1527
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      1573..2001
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1660..1679)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1870..1891
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2010..2030)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2015..2344
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2195..2330
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2257..2276
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        2392..2439
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             2458..2856
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     polyA_signal    3014..3135
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3051..3070)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3105..3124
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3184..3200)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3184..3200)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3197..3219)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3208..3224
                     /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)."
     promoter        complement(join(3232..3238,3239..3256,3257..3262))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3277..3298
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3415..3432)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3586..4174)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3666..3685)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4345..5136,5137..5205))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     4968..4987
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5206..5310)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(5385..5404)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 tgtacgggcc agatatacgc gttgacattg attattgact agttattaat agtaatcaat
       61 tacggggtca ttagttcata gcccatatat ggagttccgc gttacataac ttacggtaaa
      121 tggcccgcct ggctgaccgc ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt
      181 tcccatagta acgccaatag ggactttcca ttgacgtcaa tgggtggagt atttacggta
      241 aactgcccac ttggcagtac atcaagtgta tcatatgcca agtacgcccc ctattgacgt
      301 caatgacggt aaatggcccg cctggcatta tgcccagtac atgaccttat gggactttcc
      361 tacttggcag tacatctacg tattagtcat cgctattacc atggtgatgc ggttttggca
      421 gtacatcaat gggcgtggat agcggtttga ctcacgggga tttccaagtc tccaccccat
      481 tgacgtcaat gggagtttgt tttggcacca aaatcaacgg gactttccaa aatgtcgtaa
      541 caactccgcc ccattgacgc aaatgggcgg taggcgtgta cggtgggagg tctatataag
      601 cagagctctc tggctaacta gagaacccac tgcttactgg cttatcgaaa ttaatacgac
      661 tcactatagg gagacccaag ctggctagaa ataattttgt ttaactttaa gaaggagata
      721 tacatatgga tgtattcatg aaaggacttt caaaggccaa ggagggagtt gtggctgctg
      781 ctgagaaaac caaacagggt gtggcagaag cagcaggaaa gacaaaagag ggtgttctct
      841 atgtaggctc caaaaccaag gagggagtgg tgcatggtgt ggcaacagtg gctgagaaga
      901 ccaaagagca agtgacaaat gttggaggag cagtggtgac gggtgtgaca gcagtagccc
      961 agaagacagt ggagggagca gggagcattg cagcagccac tggctttgtc aaaaaggacc
     1021 agttgggcaa gaatgaagaa ggagccccac aggaaggaat tctggaagat atgcctgtgg
     1081 atcctgacaa tgaggcttat gaaatgcctt ctgaggaagg gtatcaagac tacgaacctg
     1141 aagcctaaga aatatctttg ctcccagttt cttgagatct gctgacagat gttccatcca
     1201 agcttcgaga agggtgggcg cgccgaccca gctttcttgt acaaagtggt gataattaat
     1261 taagatctag agggcccgtt taaacccgct gatcagcctc gactgtgcct tctagttgcc
     1321 agccatctgt tgtttgcccc tcccccgtgc cttccttgac cctggaaggt gccactccca
     1381 ctgtcctttc ctaataaaat gaggaaattg catcgcattg tctgagtagg tgtcattcta
     1441 ttctgggggg tggggtgggg caggacagca agggggagga ttgggaagac aatagcaggc
     1501 atgctgggga tgcggtgggc tctatggctt ctgaggcgga aagaaccagc tggggctcta
     1561 gggggtatcc ccacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc
     1621 gcagcgtgac cgctacactt gccagcgccc tagcgcccgc tcctttcgct ttcttccctt
     1681 cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggg ctccctttag
     1741 ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgattag ggtgatggtt
     1801 cacgtagtgg gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt
     1861 tctttaatag tggactcttg ttccaaactg gaacaacact caaccctatc tcggtctatt
     1921 cttttgattt ataagggatt ttgccgattt cggcctattg gttaaaaaat gagctgattt
     1981 aacaaaaatt taacgcgaat taattctgtg gaatgtgtgt cagttagggt gtggaaagtc
     2041 cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccag
     2101 gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta
     2161 gtcagcaacc atagtcccgc ccctaactcc gcccatcccg cccctaactc cgcccagttc
     2221 cgcccattct ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc
     2281 ctctgcctct gagctattcc agaagtagtg aggaggcttt tttggaggcc taggcttttg
     2341 caaaaagctc ccgggagctt gtatatccat tttcggatct gatcagcacg tgttgacaat
     2401 taatcatcgg catagtatat cggcatagta taatacgaca aggtgaggaa ctaaaccatg
     2461 gccaagcctt tgtctcaaga agaatccacc ctcattgaaa gagcaacggc tacaatcaac
     2521 agcatcccca tctctgaaga ctacagcgtc gccagcgcag ctctctctag cgacggccgc
     2581 atcttcactg gtgtcaatgt atatcatttt actgggggac cttgtgcaga actcgtggtg
     2641 ctgggcactg ctgctgctgc ggcagctggc aacctgactt gtatcgtcgc gatcggaaat
     2701 gagaacaggg gcatcttgag cccctgcgga cggtgccgac aggtgcttct cgatctgcat
     2761 cctgggatca aagccatagt gaaggacagt gatggacagc cgacggcagt tgggattcgt
     2821 gaattgctgc cctctggtta tgtgtgggag ggctaagcac ttcgtggccg aggagcagga
     2881 ctgacacgtg ctacgagatt tcgattccac cgccgccttc tatgaaaggt tgggcttcgg
     2941 aatcgttttc cgggacgccg gctggatgat cctccagcgc ggggatctca tgctggagtt
     3001 cttcgcccac cccaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat
     3061 cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact
     3121 catcaatgta tcttatcatg tctgtatacc gtcgacctct agctagagct tggcgtaatc
     3181 atggtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac acaacatacg
     3241 agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac tcacattaat
     3301 tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg
     3361 aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct
     3421 cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc
     3481 ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg
     3541 ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg
     3601 cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg
     3661 actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac
     3721 cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca
     3781 tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt
     3841 gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc
     3901 caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag
     3961 agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac
     4021 tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt
     4081 tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa
     4141 gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg
     4201 gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa
     4261 aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat
     4321 atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc
     4381 gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat
     4441 acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc
     4501 ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc
     4561 tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag
     4621 ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg
     4681 ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg
     4741 atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag
     4801 taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt
     4861 catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga
     4921 atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc
     4981 acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc
     5041 aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc
     5101 ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc
     5161 cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca
     5221 atattattga agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat
     5281 ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgacgt
     5341 cgacggatcg ggagatctcc cgatccccta tggtgcactc tcagtacaat ctgctctgat
     5401 gccgcatagt taagccagta tctgctccct gcttgtgtgt tggaggtcgc tgagtagtgc
     5461 gcgagcaaaa tttaagctac aacaaggcaa ggcttgaccg acaattgcat gaagaatctg
     5521 cttagggtta ggcgttttgc gctgcttcgc ga
//
