LOCUS       sequence-176939-        6268 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  expresses human beta Synuclein fused to the N-terminal part of 
            Venus.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6268)
  AUTHORS   Tenreiro S, Rosado-Ramos R, Gerhardt E, Favretto F, Magalhaes F, 
            Popova B, Becker S, Zweckstetter M, Braus GH, Outeiro TF
  TITLE     Yeast reveals similar molecular mechanisms underlying alpha- and 
            beta-synuclein toxicity.
  JOURNAL   Hum Mol Genet. 2016 Jan 15;25(2):275-90. doi: 10.1093/hmg/ddv470. 
            Epub 2015 Nov 18.
  PUBMED    26586132
REFERENCE   2  (bases 1 to 6268)
  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: 
            "10.1093/hmg/ddv470"; journalName: "Hum Mol Genet"; date: 
            "2016-01-15- 15"; volume: "25"; issue: "2"; pages: "275-90"
COMMENT     Sequence Label: VN-beta Synuclein
FEATURES             Location/Qualifiers
     source          1..6268
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(34..53)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        225..604
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        605..808
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     759..779
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     853..872
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        853..871
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             join(894..896,897..899,900..1358)
                     /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="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KLICTTGKLPVPWPTLVTTLGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITA"
     primer_bind     complement(939..960)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1200..1219)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     complement(1849..1866)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1855..2079
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2125..2553
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2212..2231)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2422..2443
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2562..2582)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2567..2896
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2747..2882
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2809..2828
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             2963..3757
                     /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="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(3017..3036)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3627..3646
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    3931..4052
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3968..3987)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4022..4041
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4101..4117)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4101..4117)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4114..4136)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4125..4141
                     /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(4149..4155,4156..4173,4174..4179))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4194..4215
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4332..4349)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4503..5091)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4583..4602)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5262..6053,6054..6122))
                     /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     5885..5904
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6123..6227)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg ccgcatagtt
       61 aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg cgagcaaaat
      121 ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc ttagggttag
      181 gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt gattattgac
      241 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg
      301 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
      361 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      421 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      481 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      541 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      601 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      661 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      721 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      781 acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca ctgcttactg
      841 gcttatcgaa attaatacga ctcactatag ggagacccaa gctggctagc accatggtga
      901 gcaagggcga ggagctgttc accggggtgg tgcccatcct ggtcgagctg gacggcgacg
      961 taaacggcca caagttcagc gtgtccggcg agggcgaggg cgatgccacc tacggcaagc
     1021 tgaccctgaa gctgatctgc accaccggca agctgcccgt gccctggccc accctcgtga
     1081 ccaccctggg ctacggcctg cagtgcttcg cccgctaccc cgaccacatg aagcagcacg
     1141 acttcttcaa gtccgccatg cccgaaggct acgtccagga gcgcaccatc ttcttcaagg
     1201 acgacggcaa ctacaagacc cgcgccgagg tgaagttcga gggcgacacc ctggtgaacc
     1261 gcatcgagct gaagggcatc gacttcaagg aggacggcaa catcctgggg cacaagctgg
     1321 agtacaacta caacagccac aacgtctata tcaccgccga caagcaaatc gatggtggcg
     1381 gtggctctgg aggtggtggg tcccttaagg acgtgttcat gaagggcctg tccatggcca
     1441 aggagggcgt cgtggcagcc gcggagaaaa ccaagcaggg ggtcaccgag gcggcggaga
     1501 agaccaagga gggcgtcctc tacgtcggaa gcaagacccg agaaggtgtg gtacaaggtg
     1561 tggcttcagt ggctgaaaaa accaaggaac aggcctcaca tctgggagga gctgtgttct
     1621 ctggggcagg gaacatcgca gcagccacag gactggtgaa gagggaggaa ttccctactg
     1681 atctgaagcc agaggaagtg gcccaggaag ctgctgaaga accactgatt gagcccctga
     1741 tggagccaga aggggagagt tatgaggacc caccccagga ggaatatcag gagtatgagc
     1801 cagaggcgta gctcgagtct agagggcccg tttaaacccg ctgatcagcc tcgactgtgc
     1861 cttctagttg ccagccatct gttgtttgcc cctcccccgt gccttccttg accctggaag
     1921 gtgccactcc cactgtcctt tcctaataaa atgaggaaat tgcatcgcat tgtctgagta
     1981 ggtgtcattc tattctgggg ggtggggtgg ggcaggacag caagggggag gattgggaag
     2041 acaatagcag gcatgctggg gatgcggtgg gctctatggc ttctgaggcg gaaagaacca
     2101 gctggggctc tagggggtat ccccacgcgc cctgtagcgg cgcattaagc gcggcgggtg
     2161 tggtggttac gcgcagcgtg accgctacac ttgccagcgc cctagcgccc gctcctttcg
     2221 ctttcttccc ttcctttctc gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg
     2281 ggctcccttt agggttccga tttagtgctt tacggcacct cgaccccaaa aaacttgatt
     2341 agggtgatgg ttcacgtagt gggccatcgc cctgatagac ggtttttcgc cctttgacgt
     2401 tggagtccac gttctttaat agtggactct tgttccaaac tggaacaaca ctcaacccta
     2461 tctcggtcta ttcttttgat ttataaggga ttttgccgat ttcggcctat tggttaaaaa
     2521 atgagctgat ttaacaaaaa tttaacgcga attaattctg tggaatgtgt gtcagttagg
     2581 gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta
     2641 gtcagcaacc aggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat
     2701 gcatctcaat tagtcagcaa ccatagtccc gcccctaact ccgcccatcc cgcccctaac
     2761 tccgcccagt tccgcccatt ctccgcccca tggctgacta atttttttta tttatgcaga
     2821 ggccgaggcc gcctctgcct ctgagctatt ccagaagtag tgaggaggct tttttggagg
     2881 cctaggcttt tgcaaaaagc tcccgggagc ttgtatatcc attttcggat ctgatcaaga
     2941 gacaggatga ggatcgtttc gcatgattga acaagatgga ttgcacgcag gttctccggc
     3001 cgcttgggtg gagaggctat tcggctatga ctgggcacaa cagacaatcg gctgctctga
     3061 tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca agaccgacct
     3121 gtccggtgcc ctgaatgaac tgcaggacga ggcagcgcgg ctatcgtggc tggccacgac
     3181 gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg actggctgct
     3241 attgggcgaa gtgccggggc aggatctcct gtcatctcac cttgctcctg ccgagaaagt
     3301 atccatcatg gctgatgcaa tgcggcggct gcatacgctt gatccggcta cctgcccatt
     3361 cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact cggatggaag ccggtcttgt
     3421 cgatcaggat gatctggacg aagagcatca ggggctcgcg ccagccgaac tgttcgccag
     3481 gctcaaggcg cgcatgcccg acggcgagga tctcgtcgtg acccatggcg atgcctgctt
     3541 gccgaatatc atggtggaaa atggccgctt ttctggattc atcgactgtg gccggctggg
     3601 tgtggcggac cgctatcagg acatagcgtt ggctacccgt gatattgctg aagagcttgg
     3661 cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc gccgctcccg attcgcagcg
     3721 catcgccttc tatcgccttc ttgacgagtt cttctgagcg ggactctggg gttcgaaatg
     3781 accgaccaag cgacgcccaa cctgccatca cgagatttcg attccaccgc cgccttctat
     3841 gaaaggttgg gcttcggaat cgttttccgg gacgccggct ggatgatcct ccagcgcggg
     3901 gatctcatgc tggagttctt cgcccacccc aacttgttta ttgcagctta taatggttac
     3961 aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact gcattctagt
     4021 tgtggtttgt ccaaactcat caatgtatct tatcatgtct gtataccgtc gacctctagc
     4081 tagagcttgg cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta tccgctcaca
     4141 attccacaca acatacgagc cggaagcata aagtgtaaag cctggggtgc ctaatgagtg
     4201 agctaactca cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg
     4261 tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc
     4321 tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta
     4381 tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag
     4441 aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg
     4501 tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg
     4561 tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg
     4621 cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga
     4681 agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc
     4741 tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt
     4801 aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact
     4861 ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg
     4921 cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt
     4981 accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt
     5041 ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct
     5101 ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg
     5161 gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt
     5221 aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt
     5281 gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc
     5341 gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg
     5401 cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc
     5461 gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg
     5521 gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca
     5581 ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga
     5641 tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct
     5701 ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg
     5761 cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca
     5821 accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata
     5881 cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct
     5941 tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact
     6001 cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa
     6061 acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc
     6121 atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga
     6181 tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga
     6241 aaagtgccac ctgacgtcga cggatcgg
//
