LOCUS       sequence-178832-        8626 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  expresses human Synphilin fused to the N-terminal part of Venus.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8626)
  AUTHORS   Dominguez-Meijide A, Parrales V, Vasili E, Gonzalez-Lizarraga F, 
            Konig A, Lazaro DF, Lannuzel A, Haik S, Del Bel E, Chehin R, 
            Raisman-Vozari R, Michel PP, Bizat N, Outeiro TF
  TITLE     Doxycycline inhibits alpha-synuclein-associated pathologies in vitro
            and in vivo.
  JOURNAL   Neurobiol Dis. 2021 Apr;151:105256. doi: 10.1016/j.nbd.2021.105256. 
            Epub 2021 Jan 8.
  PUBMED    33429042
REFERENCE   2  (bases 1 to 8626)
  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: "Neurobiol 
            Dis."; date: "2021-04"; volume: "151:105256. doi"; pages: " 
            10.1016/j.nbd.2021.105256. Epub 2021 Jan 8"
COMMENT     Sequence Label: VN-Sph1
FEATURES             Location/Qualifiers
     source          1..8626
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(2891..2908)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2897..3121
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3167..3595
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3254..3273)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3464..3485
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3604..3624)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3609..3938
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3789..3924
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3851..3870
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4005..4799
                     /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(4059..4078)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4669..4688
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4973..5094
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5010..5029)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5064..5083
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5143..5159)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5143..5159)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5156..5178)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5167..5183
                     /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(5191..5197,5198..5215,5216..5221))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5236..5257
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5374..5391)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5545..6133)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5625..5644)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6304..7095,7096..7164))
                     /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     6927..6946
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7165..7269)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(7344..7363)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        7535..7914
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        7915..8118
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     8069..8089
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     8163..8182
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        8163..8181
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             join(8204..8206,8207..8209,8210..8626,1..42)
                     /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(8249..8270)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(8510..8529)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
ORIGIN
        1 ctggagtaca actacaacag ccacaacgtc tatatcaccg ccgacaagca aatcgatggt
       61 ggcggtggct ctggaggtgg tgggtccctt aagatggaag cccctgaata ccttgatttg
      121 gatgaaattg actttagtga tgacatatct tattcagtca catcactcaa gacgatccca
      181 gaactgtgcc gaagatgtga tacgcaaaac gaagacagat cagtttctag ctctagctgg
      241 aattgtggca tctcaactct tattacaaac acgcaaaagc ccacaggaat cgctgatgtg
      301 tacagtaagt tccgcccagt gaagcgggtt tcgccactga aacatcagcc agagactctg
      361 gagaacaatg aaagtgatga ccaaaagaac cagaaagtgg ttgagtacca gaaagggggt
      421 gagtctgacc tgggccccca gcctcaggag cttggccctg gagatggagt gggcggccca
      481 ccaggtaaga gctctgagcc cagcacatcg ctgggtgaac tggagcacta cgacctcgac
      541 atggatgaga ttctggatgt gccttatatt aaatccagtc agcagcttgc ctcttttacc
      601 aaggtgactt cagaaaaaag aattttgggc ttatgcacaa ccatcaatgg cctttctggc
      661 aaagcctgct ctacaggaag ttctgagagc tcatcatcca acatggcacc attttgtgtt
      721 ctttctcccg tgaaaagccc tcacttgaga aaagcatcag ctgtcatcca cgaccagcac
      781 aagctgtcca ctgaagaaac cgagatctca cctcctctgg ttaaatgtgg ctctgcatat
      841 gagcctgaaa accagagtaa agacttccta aacaagacat ttagtgatcc tcatggtcga
      901 aaagttgaga agacaacacc agactgccag ctcagggcct tccacctaca atcctcagca
      961 gcagaatcca aaccagaaga gcaggtcagt ggcctaaacc ggaccagctc ccaaggccca
     1021 gaagaaagga gtgagtatct gaaaaaagtg aaaagcatct tgaacattgt taaagaagga
     1081 cagatctctc tcctgccaca cctagctgca gacaatctag acaaaattca cgacgaaaat
     1141 ggaaacaatc tattacatat tgcggcgtca cagggacacg cagagtgtct acagcacctc
     1201 acttctttga tgggagaaga ctgcctcaat gagcgcaaca ctgagaagtt gactccagca
     1261 ggcctggcca ttaagaatgg tcagttggag tgcgtacgct ggatggtgag cgaaacagaa
     1321 gccattgcag aactgagttg ttctaaggat tttccaagcc ttattcatta cgcaggttgc
     1381 tatggccagg aaaagattct tctgtggctt cttcagttta tgcaagaaca gggcatctcg
     1441 ttggatgaag tagaccagga tggcaacagt gccgttcacg tagcctcaca gcatggctac
     1501 cttggatgca tacagacctt ggttgaatat ggagcaaatg tcaccatgca gaaccacgct
     1561 ggggaaaagc cctcccagag cgccgagcgg caggggcaca ccctgtgctc caggtacctg
     1621 gtggtggtgg agacctgcat gtcgctggcc tctcaagtgg tgaagttaac caagcagcta
     1681 aaggaacaaa cagtagaacg tgtcacgctg cagaaccaac tccaacaatt tctagaagcc
     1741 cagaaatcag agggcaagtc actcccttct tcacccagtt cgccatcctc acctgcctcc
     1801 agaaagtccc agtggaaatc tccagatgca gatgatgatt ctgtagccaa aagcaagcca
     1861 ggagtccaag aggggattca ggttcttgga agcctgtcag cctccagccg ggctagaccc
     1921 aaagcaaaag atgaagattc tgataaaatc ttacgccagt tattgggaaa ggaaatctca
     1981 gaaaatgtct gcacccagga aaaactgtcc ttggaattcc aggatgctca ggcttcctct
     2041 agaaattcta aaaagatccc actggagaag agggaactga agttagccag gctgagacag
     2101 ctgatgcaga ggtcactgag tgagtctgac acagactcca acaactctga ggaccccaag
     2161 actaccccag tgaggaaggc tgaccgacca aggccgcagc ccattgtaga aagcgtagag
     2221 agtatggaca gcgcagaaag cctgcacctg atgattaaga aacacacctt ggcatcaggg
     2281 ggacgcaggt ttcctttcag catcaaggcc tccaaatccc tggatggcca cagcccatct
     2341 cccacctcag agagcagcga accagactta gaatcccagt atccaggctc agggagtatt
     2401 cctccaaacc agccctctgg tgaccctcag cagcccagcc ctgacagtac tgctgcccag
     2461 aaagttgcca caagtcccaa gagtgccctc aagtctccat cttccaagcg taggacatct
     2521 cagaacttaa aactgagagt tacctttgag gagcctgtgg tgcagatgga gcagcctagc
     2581 cttgaactga atggagaaaa agacaaagat aagggaagga ctctccagcg gacctccaca
     2641 agtaacgaat cgggggatca actgaaaagg ccttttggag cctttcgatc tatcatggag
     2701 acactaagtg gcaaccaaaa caataataat aactaccagg cagccaacca gctgaaaacc
     2761 tctacattgc ccttgacctc acttgggagg aagacagatg ccaagggaaa ccctgccagc
     2821 tccgctagca aaggaaagaa taaggcagca taactcgagt ctagagggcc cgtttaaacc
     2881 cgctgatcag cctcgactgt gccttctagt tgccagccat ctgttgtttg cccctccccc
     2941 gtgccttcct tgaccctgga aggtgccact cccactgtcc tttcctaata aaatgaggaa
     3001 attgcatcgc attgtctgag taggtgtcat tctattctgg ggggtggggt ggggcaggac
     3061 agcaaggggg aggattggga agacaatagc aggcatgctg gggatgcggt gggctctatg
     3121 gcttctgagg cggaaagaac cagctggggc tctagggggt atccccacgc gccctgtagc
     3181 ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgccagc
     3241 gccctagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt
     3301 ccccgtcaag ctctaaatcg ggggctccct ttagggttcc gatttagtgc tttacggcac
     3361 ctcgacccca aaaaacttga ttagggtgat ggttcacgta gtgggccatc gccctgatag
     3421 acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact cttgttccaa
     3481 actggaacaa cactcaaccc tatctcggtc tattcttttg atttataagg gattttgccg
     3541 atttcggcct attggttaaa aaatgagctg atttaacaaa aatttaacgc gaattaattc
     3601 tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg ctccccagca ggcagaagta
     3661 tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag
     3721 caggcagaag tatgcaaagc atgcatctca attagtcagc aaccatagtc ccgcccctaa
     3781 ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc catggctgac
     3841 taattttttt tatttatgca gaggccgagg ccgcctctgc ctctgagcta ttccagaagt
     3901 agtgaggagg cttttttgga ggcctaggct tttgcaaaaa gctcccggga gcttgtatat
     3961 ccattttcgg atctgatcaa gagacaggat gaggatcgtt tcgcatgatt gaacaagatg
     4021 gattgcacgc aggttctccg gccgcttggg tggagaggct attcggctat gactgggcac
     4081 aacagacaat cggctgctct gatgccgccg tgttccggct gtcagcgcag gggcgcccgg
     4141 ttctttttgt caagaccgac ctgtccggtg ccctgaatga actgcaggac gaggcagcgc
     4201 ggctatcgtg gctggccacg acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg
     4261 aagcgggaag ggactggctg ctattgggcg aagtgccggg gcaggatctc ctgtcatctc
     4321 accttgctcc tgccgagaaa gtatccatca tggctgatgc aatgcggcgg ctgcatacgc
     4381 ttgatccggc tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta
     4441 ctcggatgga agccggtctt gtcgatcagg atgatctgga cgaagagcat caggggctcg
     4501 cgccagccga actgttcgcc aggctcaagg cgcgcatgcc cgacggcgag gatctcgtcg
     4561 tgacccatgg cgatgcctgc ttgccgaata tcatggtgga aaatggccgc ttttctggat
     4621 tcatcgactg tggccggctg ggtgtggcgg accgctatca ggacatagcg ttggctaccc
     4681 gtgatattgc tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg ctttacggta
     4741 tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct tcttgacgag ttcttctgag
     4801 cgggactctg gggttcgaaa tgaccgacca agcgacgccc aacctgccat cacgagattt
     4861 cgattccacc gccgccttct atgaaaggtt gggcttcgga atcgttttcc gggacgccgg
     4921 ctggatgatc ctccagcgcg gggatctcat gctggagttc ttcgcccacc ccaacttgtt
     4981 tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc
     5041 atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttatcatgt
     5101 ctgtataccg tcgacctcta gctagagctt ggcgtaatca tggtcatagc tgtttcctgt
     5161 gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca taaagtgtaa
     5221 agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct cactgcccgc
     5281 tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag
     5341 aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt
     5401 cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga
     5461 atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg
     5521 taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa
     5581 aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt
     5641 tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct
     5701 gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct
     5761 cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc
     5821 cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt
     5881 atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc
     5941 tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat
     6001 ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa
     6061 acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa
     6121 aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga
     6181 aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct
     6241 tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga
     6301 cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc
     6361 catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg
     6421 ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat
     6481 aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat
     6541 ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg
     6601 caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc
     6661 attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa
     6721 agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc
     6781 actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt
     6841 ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag
     6901 ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt
     6961 gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag
     7021 atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac
     7081 cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc
     7141 gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca
     7201 gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg
     7261 ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc gacggatcgg gagatctccc
     7321 gatcccctat ggtgcactct cagtacaatc tgctctgatg ccgcatagtt aagccagtat
     7381 ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg cgagcaaaat ttaagctaca
     7441 acaaggcaag gcttgaccga caattgcatg aagaatctgc ttagggttag gcgttttgcg
     7501 ctgcttcgcg atgtacgggc cagatatacg cgttgacatt gattattgac tagttattaa
     7561 tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg cgttacataa
     7621 cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt gacgtcaata
     7681 atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca atgggtggag
     7741 tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc aagtacgccc
     7801 cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta catgacctta
     7861 tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac catggtgatg
     7921 cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg atttccaagt
     7981 ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg ggactttcca
     8041 aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt acggtgggag
     8101 gtctatataa gcagagctct ctggctaact agagaaccca ctgcttactg gcttatcgaa
     8161 attaatacga ctcactatag ggagacccaa gctggctagc accatggtga gcaagggcga
     8221 ggagctgttc accggggtgg tgcccatcct ggtcgagctg gacggcgacg taaacggcca
     8281 caagttcagc gtgtccggcg agggcgaggg cgatgccacc tacggcaagc tgaccctgaa
     8341 gctgatctgc accaccggca agctgcccgt gccctggccc accctcgtga ccaccctggg
     8401 ctacggcctg cagtgcttcg cccgctaccc cgaccacatg aagcagcacg acttcttcaa
     8461 gtccgccatg cccgaaggct acgtccagga gcgcaccatc ttcttcaagg acgacggcaa
     8521 ctacaagacc cgcgccgagg tgaagttcga gggcgacacc ctggtgaacc gcatcgagct
     8581 gaagggcatc gacttcaagg aggacggcaa catcctgggg cacaag
//
