LOCUS       sequence-207251-        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 A30P
FEATURES             Location/Qualifiers
     source          1..5552
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        32..411
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        412..615
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     566..586
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     660..679
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        660..678
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             704..726
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     protein_bind    complement(1234..1258)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(1305..1322)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1311..1535
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      1581..2009
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1668..1687)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1878..1899
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2018..2038)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2023..2352
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2203..2338
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2265..2284
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        2400..2447
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             2466..2864
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     polyA_signal    3022..3143
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3059..3078)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3113..3132
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3192..3208)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3192..3208)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3205..3227)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3216..3232
                     /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(3240..3246,3247..3264,3265..3270))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3285..3306
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3423..3440)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3594..4182)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3674..3693)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4353..5144,5145..5213))
                     /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     4976..4995
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5214..5318)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(5393..5412)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 cttcgcgatg tacgggccag atatacgcgt tgacattgat tattgactag ttattaatag
       61 taatcaatta cggggtcatt agttcatagc ccatatatgg agttccgcgt tacataactt
      121 acggtaaatg gcccgcctgg ctgaccgccc aacgaccccc gcccattgac gtcaataatg
      181 acgtatgttc ccatagtaac gccaataggg actttccatt gacgtcaatg ggtggagtat
      241 ttacggtaaa ctgcccactt ggcagtacat caagtgtatc atatgccaag tacgccccct
      301 attgacgtca atgacggtaa atggcccgcc tggcattatg cccagtacat gaccttatgg
      361 gactttccta cttggcagta catctacgta ttagtcatcg ctattaccat ggtgatgcgg
      421 ttttggcagt acatcaatgg gcgtggatag cggtttgact cacggggatt tccaagtctc
      481 caccccattg acgtcaatgg gagtttgttt tggcaccaaa atcaacggga ctttccaaaa
      541 tgtcgtaaca actccgcccc attgacgcaa atgggcggta ggcgtgtacg gtgggaggtc
      601 tatataagca gagctctctg gctaactaga gaacccactg cttactggct tatcgaaatt
      661 aatacgactc actataggga gacccaagct ggctagaaat aattttgttt aactttaaga
      721 aggagatata catatggatg tattcatgaa aggactttca aaggccaagg agggagttgt
      781 ggctgctgct gagaaaacca aacagggtgt ggcagaagca ccaggaaaga caaaagaggg
      841 tgttctctat gtaggctcca aaaccaagga gggagtggtg catggtgtgg caacagtggc
      901 tgagaagacc aaagagcaag tgacaaatgt tggaggagca gtggtgacgg gtgtgacagc
      961 agtagcccag aagacagtgg agggagcagg gagcattgca gcagccactg gctttgtcaa
     1021 aaaggaccag ttgggcaaga atgaagaagg agccccacag gaaggaattc tggaagatat
     1081 gcctgtggat cctgacaatg aggcttatga aatgccttct gaggaagggt atcaagacta
     1141 cgaacctgaa gcctaagaaa tatctttgct cccagtttct tgagatctgc tgacagatgt
     1201 tccatccaag cttcgagaag ggtgggcgcg ccgacccagc tttcttgtac aaagtggtga
     1261 taattaatta agatctagag ggcccgttta aacccgctga tcagcctcga ctgtgccttc
     1321 tagttgccag ccatctgttg tttgcccctc ccccgtgcct tccttgaccc tggaaggtgc
     1381 cactcccact gtcctttcct aataaaatga ggaaattgca tcgcattgtc tgagtaggtg
     1441 tcattctatt ctggggggtg gggtggggca ggacagcaag ggggaggatt gggaagacaa
     1501 tagcaggcat gctggggatg cggtgggctc tatggcttct gaggcggaaa gaaccagctg
     1561 gggctctagg gggtatcccc acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt
     1621 ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt
     1681 cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcgggggct
     1741 ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgattaggg
     1801 tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga
     1861 gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca accctatctc
     1921 ggtctattct tttgatttat aagggatttt gccgatttcg gcctattggt taaaaaatga
     1981 gctgatttaa caaaaattta acgcgaatta attctgtgga atgtgtgtca gttagggtgt
     2041 ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca
     2101 gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat
     2161 ctcaattagt cagcaaccat agtcccgccc ctaactccgc ccatcccgcc cctaactccg
     2221 cccagttccg cccattctcc gccccatggc tgactaattt tttttattta tgcagaggcc
     2281 gaggccgcct ctgcctctga gctattccag aagtagtgag gaggcttttt tggaggccta
     2341 ggcttttgca aaaagctccc gggagcttgt atatccattt tcggatctga tcagcacgtg
     2401 ttgacaatta atcatcggca tagtatatcg gcatagtata atacgacaag gtgaggaact
     2461 aaaccatggc caagcctttg tctcaagaag aatccaccct cattgaaaga gcaacggcta
     2521 caatcaacag catccccatc tctgaagact acagcgtcgc cagcgcagct ctctctagcg
     2581 acggccgcat cttcactggt gtcaatgtat atcattttac tgggggacct tgtgcagaac
     2641 tcgtggtgct gggcactgct gctgctgcgg cagctggcaa cctgacttgt atcgtcgcga
     2701 tcggaaatga gaacaggggc atcttgagcc cctgcggacg gtgccgacag gtgcttctcg
     2761 atctgcatcc tgggatcaaa gccatagtga aggacagtga tggacagccg acggcagttg
     2821 ggattcgtga attgctgccc tctggttatg tgtgggaggg ctaagcactt cgtggccgag
     2881 gagcaggact gacacgtgct acgagatttc gattccaccg ccgccttcta tgaaaggttg
     2941 ggcttcggaa tcgttttccg ggacgccggc tggatgatcc tccagcgcgg ggatctcatg
     3001 ctggagttct tcgcccaccc caacttgttt attgcagctt ataatggtta caaataaagc
     3061 aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg
     3121 tccaaactca tcaatgtatc ttatcatgtc tgtataccgt cgacctctag ctagagcttg
     3181 gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac
     3241 aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc
     3301 acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg
     3361 cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct
     3421 tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac
     3481 tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga
     3541 gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat
     3601 aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac
     3661 ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct
     3721 gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg
     3781 ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg
     3841 ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt
     3901 cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg
     3961 attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac
     4021 ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga
     4081 aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt
     4141 gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt
     4201 tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga
     4261 ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc
     4321 taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct
     4381 atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata
     4441 actacgatac gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca
     4501 cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga
     4561 agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga
     4621 gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg
     4681 gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga
     4741 gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt
     4801 gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct
     4861 cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca
     4921 ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat
     4981 accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga
     5041 aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc
     5101 aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg
     5161 caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc
     5221 ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt
     5281 gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca
     5341 cctgacgtcg acggatcggg agatctcccg atcccctatg gtgcactctc agtacaatct
     5401 gctctgatgc cgcatagtta agccagtatc tgctccctgc ttgtgtgttg gaggtcgctg
     5461 agtagtgcgc gagcaaaatt taagctacaa caaggcaagg cttgaccgac aattgcatga
     5521 agaatctgct tagggttagg cgttttgcgc tg
//
