LOCUS       sequence-181603-        7541 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Retroviral overexpression vector for alpha-Syn bimolecular 
            fluorescence complementation (BiFC).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7541)
  AUTHORS   Rousseaux MW, de Haro M, Lasagna-Reeves CA, De Maio A, Park J, 
            Jafar-Nejad P, Al-Ramahi I, Sharma A, See L, Lu N, Vilanova-Velez L,
            Klisch TJ, Westbrook TF, Troncoso JC, Botas J, Zoghbi HY
  TITLE     TRIM28 regulates the nuclear accumulation and toxicity of both 
            alpha-synuclein and tau.
  JOURNAL   Elife. 2016 Oct 25;5. pii: e19809. doi: 10.7554/eLife.19809.
  PUBMED    27779468
REFERENCE   2  (bases 1 to 7541)
  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"; journalName: "Elife."; 
            date: "2016-10-25"; volume: "5. pii: e19809. doi"; pages: " 
            10.7554/eLife.19809"
COMMENT     Sequence Label: pBABE-alpha-Syn-nYFP
FEATURES             Location/Qualifiers
     source          1..7541
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     LTR             22..491
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     misc_feature    556..755
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     CDS             956..1372
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     1302..1324
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     1340..1356
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     primer_bind     complement(1409..1428)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        1600..1979
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1980..2183
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2134..2154
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     2180..2204
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     promoter        2414..2432
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     2414..2431
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     2445..2464
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     protein_bind    2510..2534
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(2991..3015)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             join(3077..3079,3080..3538)
                     /codon_start=1
                     /product="improved N-terminal fragment of mVenus for use in
                     bimolecular fluorescence complementation (BiFC) (Kodama and
                     Hu, 2010)"
                     /label=VN155(I152L)
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     LICTTGKLPVPWPTLVTTLGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITA"
     primer_bind     complement(3119..3140)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3380..3399)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     complement(3606..3629)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     3969..3988
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     CDS             4050..4853
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MGSAIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQ
                     GRPVLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQD
                     LLSSHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDE
                     EHQGLAPAELFARLKARMPDGDDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQ
                     DIALATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(4113..4132)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4723..4742
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     LTR             5049..5518
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     primer_bind     complement(5648..5665)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5819..6407)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5899..5918)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6567..7358,7359..7427))
                     /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     7190..7209
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7428..7532)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ccgaaaagtg ccacctgcag cctgaatatg ggccaaacag gatatctgtg gtaagcagtt
       61 cctgccccgg ctcagggcca agaacagatg gaacagctga atatgggcca aacaggatat
      121 ctgtggtaag cagttcctgc cccggctcag ggccaagaac agatggtccc cagatgcggt
      181 ccagccctca gcagtttcta gagaaccatc agatgtttcc agggtgcccc aaggacctga
      241 aatgaccctg tgccttattt gaactaacca atcagttcgc ttctcgcttc tgttcgcgcg
      301 cttctgctcc ccgagctcaa taaaagagcc cacaacccct cactcggggc gccagtcctc
      361 cgattgactg agtcgcccgg gtacccgtgt atccaataaa ccctcttgca gttgcatccg
      421 acttgtggtc tcgctgttcc ttgggagggt ctcctctgag tgattgacta cccgtcagcg
      481 ggggtctttc atttgggggc tcgtccggga tcgggagacc cctgcccagg gaccaccgac
      541 ccaccaccgg gaggcaagct ggccagcaac ttatctgtgt ctgtccgatt gtctagtgtc
      601 tatgactgat tttatgcgcc tgcgtcggta ctagttagct aactagctct gtatctggcg
      661 gacccgtggt ggaactgacg agttctgaac acccggccgc aaccctggga gacgtcccag
      721 ggactttggg ggccgttttt gtggcccgac ctgaggaagg gagtcgatgt ggaatccgac
      781 cccgtcagga tatgtggttc tggtaggaga cgagaaccta aaacagttcc cgcctccgtc
      841 tgaatttttg ctttcggttt ggaaccgaag ccgcgcgtct tgtctgctgc agcgctgcag
      901 catcgttctg tgttgtctct gtctgactgt gtttctgtat ttgtctgaaa attagggcca
      961 gactgttacc actcccttaa gtttgacctt aggtcactgg aaagatgtcg agcggatcgc
     1021 tcacaaccag tcggtagatg tcaagaagag acgttgggtt accttctgct ctgcagaatg
     1081 gccaaccttt aacgtcggat ggccgcgaga cggcaccttt aaccgagacc tcatcaccca
     1141 ggttaagatc aaggtctttt cacctggccc gcatggacac ccagaccagg tcccctacat
     1201 cgtgacctgg gaagccttgg cttttgaccc ccctccctgg gtcaagccct ttgtacaccc
     1261 taagcctccg cctcctcttc ctccatccgc cccgtctctc ccccttgaac ctcctcgttc
     1321 gaccccgcct cgatcctccc tttatccagc cctcactcct tctctaggcg ccggccggat
     1381 ctcccgatcc cctatggtgc actctcagta caatctgctc tgatgccgca tagttaagcc
     1441 agtatctgct ccctgcttgt gtgttggagg tcgctgagta gtgcgcgagc aaaatttaag
     1501 ctacaacaag gcaaggcttg accgacaatt gcatgaagaa tctgcttagg gttaggcgtt
     1561 ttgcgctgct tcgcgatgta cgggccagat atacgcgttg acattgatta ttgactagtt
     1621 attaatagta atcaattacg gggtcattag ttcatagccc atatatggag ttccgcgtta
     1681 cataacttac ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc ccattgacgt
     1741 caataatgac gtatgttccc atagtaacgc caatagggac tttccattga cgtcaatggg
     1801 tggagtattt acggtaaact gcccacttgg cagtacatca agtgtatcat atgccaagta
     1861 cgccccctat tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc cagtacatga
     1921 ccttatggga ctttcctact tggcagtaca tctacgtatt agtcatcgct attaccatgg
     1981 tgatgcggtt ttggcagtac atcaatgggc gtggatagcg gtttgactca cggggatttc
     2041 caagtctcca ccccattgac gtcaatggga gtttgttttg gcaccaaaat caacgggact
     2101 ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat gggcggtagg cgtgtacggt
     2161 gggaggtcta tataagcaga gctcgtttag tgaaccgtca gatcgcctgg agacgccatc
     2221 cacgctgttt tgacctccat agaagacacc gggaccgatc cagcctccgc ggccgggaac
     2281 ggtgcattgg aacgcggatt ccccgtgcca agagtgacgt aagtaccgcc tatagagtct
     2341 ataggcccac ccccttggct tcgttagaac gcggctacaa ttaatacata accttatgta
     2401 tcatacacat acgatttagg tgacactata gaataacatc cactttgcct ttctctccac
     2461 aggtgtccac tcccaggtcc aactgcacct cggttctatc gattgatcaa caagtttgta
     2521 caaaaaagca ggctccgcgg ccgccccctt caccatggat gtattcatga aaggactttc
     2581 aaaggccaag gagggagttg tggctgctgc tgagaaaacc aaacagggtg tggcagaagc
     2641 agcaggaaag acaaaagagg gtgttctcta tgtaggctcc aaaaccaagg agggagtggt
     2701 gcatggtgtg gcaacagtgg ctgagaagac caaagagcaa gtgacaaatg ttggaggagc
     2761 agtggtgacg ggtgtgacag cagtagccca gaagacagtg gagggagcag ggagcattgc
     2821 agcagccact ggctttgtca aaaaggacca gttgggcaag aatgaagaag gagccccaca
     2881 ggaaggaatt ctggaagata tgcctgtgga tcctgacaat gaggcttatg aaatgccttc
     2941 tgaggaaggg tatcaagact acgaacctga agccaagggt gggcgcgccg acccagcttt
     3001 cttgtacaaa gtggttgatc cccggggatc ctctagagtc gacggcggag gaggttcagg
     3061 aggcggaggc gcgccagtga gcaagggcga ggagctgttc accggggtgg tgcccatcct
     3121 ggtcgagctg gacggcgacg taaacggcca caagttcagc gtgtccggcg agggcgaggg
     3181 cgatgccacc tacggcaagc tgaccctgaa gctgatctgc accaccggca agctgcccgt
     3241 gccctggccc accctcgtga ccaccctggg ctacggcctg cagtgcttcg cccgctaccc
     3301 cgaccacatg aagcagcacg acttcttcaa gtccgccatg cccgaaggct acgtccagga
     3361 gcgcaccatc ttcttcaagg acgacggcaa ctacaagacc cgcgccgagg tgaagttcga
     3421 gggcgacacc ctggtgaacc gcatcgagct gaagggcatc gacttcaagg aggacggcaa
     3481 catcctgggg cacaagctgg agtacaacta caacagccac aacgtctata tcactgccta
     3541 gttaacaacc tcgagagatc taattctacc gggtagggga ggcgcttttc ccaaggcagt
     3601 ctggagcatg cgctttagca gccccgctgg gcacttggcg ctacacaagt ggcctctggc
     3661 ctcgcacaca ttccacatcc accggtaggc gccaaccggc tccgttcttt ggtggcccct
     3721 tcgcgccacc ttctactcct cccctagtca ggaagttccc ccccgccccg cagctcgcgt
     3781 cgtgcaggac gtgacaaatg gaagtagcac gtctcactag tctcgtgcag atggacagca
     3841 ccgctgagca atggaagcgg gtaggccttt ggggcagcgg ccaatagcag ctttgctcct
     3901 tcgctttctg ggctcagagg ctgggaaggg gtgggtccgg gggccccgaa ggtcctccgg
     3961 aggcccggca ttctgcacgc ttcaaaagcg cacgtctgcc gcgctgttct cctcttcctc
     4021 atctccgggc ctttcgacct gcagccaata tgggatcggc cattgaacaa gatggattgc
     4081 acgcaggttc tccggccgct tgggtggaga ggctattcgg ctatgactgg gcacaacaga
     4141 caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt
     4201 ttgtcaagac cgacctgtcc ggtgccctga atgaactgca ggacgaggca gcgcggctat
     4261 cgtggctggc cacgacgggc gttccttgcg cagctgtgct cgacgttgtc actgaagcgg
     4321 gaagggactg gctgctattg ggcgaagtgc cggggcagga tctcctgtca tctcaccttg
     4381 ctcctgccga gaaagtatcc atcatggctg atgcaatgcg gcggctgcat acgcttgatc
     4441 cggctacctg cccattcgac caccaagcga aacatcgcat cgagcgagca cgtactcgga
     4501 tggaagccgg tcttgtcgat caggatgatc tggacgaaga gcatcagggg ctcgcgccag
     4561 ccgaactgtt cgccaggctc aaggcgcgca tgcccgacgg cgatgatctc gtcgtgaccc
     4621 atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct ggattcatcg
     4681 actgtggccg gctgggtgtg gcggaccgct atcaggacat agcgttggct acccgtgata
     4741 ttgctgaaga gcttggcggc gaatgggctg accgcttcct cgtgctttac ggtatcgccg
     4801 ctcccgattc gcagcgcatc gccttctatc gccttcttga cgagttcttc tgaggggatc
     4861 cgtcgacctg cagccaagct tatcgataaa ataaaagatt ttatttagtc tccagaaaaa
     4921 ggggggaatg aaagacccca cctgtaggtt tggcaagcta gcttaagtaa cgccattttg
     4981 caaggcatgg aaaaatacat aactgagaat agagaagttc agatcaaggt caggaacaga
     5041 tggaacagct gaatatgggc caaacaggat atctgtggta agcagttcct gccccggctc
     5101 agggccaaga acagatggaa cagctgaata tgggccaaac aggatatctg tggtaagcag
     5161 ttcctgcccc ggctcagggc caagaacaga tggtccccag atgcggtcca gccctcagca
     5221 gtttctagag aaccatcaga tgtttccagg gtgccccaag gacctgaaat gaccctgtgc
     5281 cttatttgaa ctaaccaatc agttcgcttc tcgcttctgt tcgcgcgctt ctgctccccg
     5341 agctcaataa aagagcccac aacccctcac tcggggcgcc agtcctccga ttgactgagt
     5401 cgcccgggta cccgtgtatc caataaaccc tcttgcagtt gcatccgact tgtggtctcg
     5461 ctgttccttg ggagggtctc ctctgagtga ttgactaccc gtcagcgggg gtctttcaca
     5521 tgcagcatgt atcaaaatta atttggtttt ttttcttaag tatttacatt aaatggccat
     5581 agttgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt
     5641 ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag
     5701 ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca
     5761 tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt
     5821 tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc
     5881 gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct
     5941 ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg
     6001 tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca
     6061 agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact
     6121 atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta
     6181 acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta
     6241 actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct
     6301 tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt
     6361 tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga
     6421 tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca
     6481 tgagattatc aaaaaggatc ttcacctaga tccttttgcg gccgcaaatc aatctaaagt
     6541 atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca
     6601 gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg
     6661 atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca
     6721 ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt
     6781 cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt
     6841 agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca
     6901 cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca
     6961 tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga
     7021 agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact
     7081 gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga
     7141 gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg
     7201 ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc
     7261 tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga
     7321 tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat
     7381 gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt
     7441 caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt
     7501 atttagaaaa ataaacaaat aggggttccg cgcacatttc c
//
