LOCUS       sequence-207254-        5552 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of mutant 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 E46K
FEATURES             Location/Qualifiers
     source          1..5552
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        45..424
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        425..628
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     579..599
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     673..692
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        673..691
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             717..739
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     protein_bind    complement(1247..1271)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(1318..1335)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1324..1548
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      1594..2022
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1681..1700)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1891..1912
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2031..2051)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2036..2365
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2216..2351
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2278..2297
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        2413..2460
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             2479..2877
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     polyA_signal    3035..3156
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3072..3091)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3126..3145
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3205..3221)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3205..3221)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3218..3240)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3229..3245
                     /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(3253..3259,3260..3277,3278..3283))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3298..3319
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3436..3453)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3607..4195)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3687..3706)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4366..5157,5158..5226))
                     /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     4989..5008
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5227..5331)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(5406..5425)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt gattattgac
       61 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg
      121 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
      181 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      241 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      301 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      361 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      421 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      481 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      541 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      601 acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca ctgcttactg
      661 gcttatcgaa attaatacga ctcactatag ggagacccaa gctggctaga aataattttg
      721 tttaacttta agaaggagat atacatatgg atgtattcat gaaaggactt tcaaaggcca
      781 aggagggagt tgtggctgct gctgagaaaa ccaaacaggg tgtggcagaa gcagcaggaa
      841 agacaaaaga gggtgttctc tatgtaggct ccaaaaccaa gaagggagtg gtgcatggtg
      901 tggcaacagt ggctgagaag accaaagagc aagtgacaaa tgttggagga gcagtggtga
      961 cgggtgtgac agcagtagcc cagaagacag tggagggagc agggagcatt gcagcagcca
     1021 ctggctttgt caaaaaggac cagttgggca agaatgaaga aggagcccca caggaaggaa
     1081 ttctggaaga tatgcctgtg gatcctgaca atgaggctta tgaaatgcct tctgaggaag
     1141 ggtatcaaga ctacgaacct gaagcctaag aaatatcttt gctcccagtt tcttgagatc
     1201 tgctgacaga tgttccatcc aagcttcgag aagggtgggc gcgccgaccc agctttcttg
     1261 tacaaagtgg tgataattaa ttaagatcta gagggcccgt ttaaacccgc tgatcagcct
     1321 cgactgtgcc ttctagttgc cagccatctg ttgtttgccc ctcccccgtg ccttccttga
     1381 ccctggaagg tgccactccc actgtccttt cctaataaaa tgaggaaatt gcatcgcatt
     1441 gtctgagtag gtgtcattct attctggggg gtggggtggg gcaggacagc aagggggagg
     1501 attgggaaga caatagcagg catgctgggg atgcggtggg ctctatggct tctgaggcgg
     1561 aaagaaccag ctggggctct agggggtatc cccacgcgcc ctgtagcggc gcattaagcg
     1621 cggcgggtgt ggtggttacg cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg
     1681 ctcctttcgc tttcttccct tcctttctcg ccacgttcgc cggctttccc cgtcaagctc
     1741 taaatcgggg gctcccttta gggttccgat ttagtgcttt acggcacctc gaccccaaaa
     1801 aacttgatta gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc
     1861 ctttgacgtt ggagtccacg ttctttaata gtggactctt gttccaaact ggaacaacac
     1921 tcaaccctat ctcggtctat tcttttgatt tataagggat tttgccgatt tcggcctatt
     1981 ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa ttaattctgt ggaatgtgtg
     2041 tcagttaggg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca
     2101 tctcaattag tcagcaacca ggtgtggaaa gtccccaggc tccccagcag gcagaagtat
     2161 gcaaagcatg catctcaatt agtcagcaac catagtcccg cccctaactc cgcccatccc
     2221 gcccctaact ccgcccagtt ccgcccattc tccgccccat ggctgactaa ttttttttat
     2281 ttatgcagag gccgaggccg cctctgcctc tgagctattc cagaagtagt gaggaggctt
     2341 ttttggaggc ctaggctttt gcaaaaagct cccgggagct tgtatatcca ttttcggatc
     2401 tgatcagcac gtgttgacaa ttaatcatcg gcatagtata tcggcatagt ataatacgac
     2461 aaggtgagga actaaaccat ggccaagcct ttgtctcaag aagaatccac cctcattgaa
     2521 agagcaacgg ctacaatcaa cagcatcccc atctctgaag actacagcgt cgccagcgca
     2581 gctctctcta gcgacggccg catcttcact ggtgtcaatg tatatcattt tactggggga
     2641 ccttgtgcag aactcgtggt gctgggcact gctgctgctg cggcagctgg caacctgact
     2701 tgtatcgtcg cgatcggaaa tgagaacagg ggcatcttga gcccctgcgg acggtgccga
     2761 caggtgcttc tcgatctgca tcctgggatc aaagccatag tgaaggacag tgatggacag
     2821 ccgacggcag ttgggattcg tgaattgctg ccctctggtt atgtgtggga gggctaagca
     2881 cttcgtggcc gaggagcagg actgacacgt gctacgagat ttcgattcca ccgccgcctt
     2941 ctatgaaagg ttgggcttcg gaatcgtttt ccgggacgcc ggctggatga tcctccagcg
     3001 cggggatctc atgctggagt tcttcgccca ccccaacttg tttattgcag cttataatgg
     3061 ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc
     3121 tagttgtggt ttgtccaaac tcatcaatgt atcttatcat gtctgtatac cgtcgacctc
     3181 tagctagagc ttggcgtaat catggtcata gctgtttcct gtgtgaaatt gttatccgct
     3241 cacaattcca cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg
     3301 agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct
     3361 gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg
     3421 gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc
     3481 ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg
     3541 aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct
     3601 ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca
     3661 gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct
     3721 cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc
     3781 gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt
     3841 tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc
     3901 cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc
     3961 cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg
     4021 gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc
     4081 agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag
     4141 cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga
     4201 tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat
     4261 tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag
     4321 ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat
     4381 cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc
     4441 cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat
     4501 accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag
     4561 ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg
     4621 ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc
     4681 tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca
     4741 acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg
     4801 tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc
     4861 actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta
     4921 ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc
     4981 aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg
     5041 ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc
     5101 cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc
     5161 aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat
     5221 actcatactc ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag
     5281 cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc
     5341 ccgaaaagtg ccacctgacg tcgacggatc gggagatctc ccgatcccct atggtgcact
     5401 ctcagtacaa tctgctctga tgccgcatag ttaagccagt atctgctccc tgcttgtgtg
     5461 ttggaggtcg ctgagtagtg cgcgagcaaa atttaagcta caacaaggca aggcttgacc
     5521 gacaattgca tgaagaatct gcttagggtt ag
//
