LOCUS       sequence-178646-        8363 bp    DNA     circular SYN 15-APR-2025
DEFINITION  expresses human Synphilin fused to the C-terminal part of Venus.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8363)
  AUTHORS   Lazaro DF, Amen T, Gerhardt E, Santos PI, Milovanovic D, Hoglinger 
            G, Outeiro TF
  TITLE     Synphilin-1 as a modulator of aSyn assembly
  JOURNAL   bioRxiv 2024.03.05.583522
REFERENCE   2  (bases 1 to 8363)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "bioRxiv 
            2024.03.05.583522"
COMMENT     Sequence Label: Sph1-VC
FEATURES             Location/Qualifiers
     source          1..8363
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     11..32
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(151..171)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        156..485
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      336..471
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     398..417
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             552..1346
                     /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(606..625)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     1216..1235
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    1520..1641
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1557..1576)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1611..1630
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(1690..1706)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1690..1706)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1703..1725)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1714..1730
                     /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(1738..1744,1745..1762,1763..1768))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1783..1804
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1921..1938)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2092..2680)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2172..2191)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2851..3642,3643..3711))
                     /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     3474..3493
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3712..3816)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(3891..3910)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        4082..4461
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4462..4665
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     4616..4636
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     4710..4729
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        4710..4728
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     7703..7724
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(7801..7818)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    7807..8031
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      join(8077..8363,1..142)
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(8164..8183)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 ttctttaata gtggactctt gttccaaact ggaacaacac tcaaccctat ctcggtctat
       61 tcttttgatt tataagggat tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt
      121 taacaaaaat ttaacgcgaa ttaattctgt ggaatgtgtg tcagttaggg tgtggaaagt
      181 ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca
      241 ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt
      301 agtcagcaac catagtcccg cccctaactc cgcccatccc gcccctaact ccgcccagtt
      361 ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg
      421 cctctgcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc ctaggctttt
      481 gcaaaaagct cccgggagct tgtatatcca ttttcggatc tgatcaagag acaggatgag
      541 gatcgtttcg catgattgaa caagatggat tgcacgcagg ttctccggcc gcttgggtgg
      601 agaggctatt cggctatgac tgggcacaac agacaatcgg ctgctctgat gccgccgtgt
      661 tccggctgtc agcgcagggg cgcccggttc tttttgtcaa gaccgacctg tccggtgccc
      721 tgaatgaact gcaggacgag gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt
      781 gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga ctggctgcta ttgggcgaag
      841 tgccggggca ggatctcctg tcatctcacc ttgctcctgc cgagaaagta tccatcatgg
      901 ctgatgcaat gcggcggctg catacgcttg atccggctac ctgcccattc gaccaccaag
      961 cgaaacatcg catcgagcga gcacgtactc ggatggaagc cggtcttgtc gatcaggatg
     1021 atctggacga agagcatcag gggctcgcgc cagccgaact gttcgccagg ctcaaggcgc
     1081 gcatgcccga cggcgaggat ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca
     1141 tggtggaaaa tggccgcttt tctggattca tcgactgtgg ccggctgggt gtggcggacc
     1201 gctatcagga catagcgttg gctacccgtg atattgctga agagcttggc ggcgaatggg
     1261 ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc atcgccttct
     1321 atcgccttct tgacgagttc ttctgagcgg gactctgggg ttcgaaatga ccgaccaagc
     1381 gacgcccaac ctgccatcac gagatttcga ttccaccgcc gccttctatg aaaggttggg
     1441 cttcggaatc gttttccggg acgccggctg gatgatcctc cagcgcgggg atctcatgct
     1501 ggagttcttc gcccacccca acttgtttat tgcagcttat aatggttaca aataaagcaa
     1561 tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc
     1621 caaactcatc aatgtatctt atcatgtctg tataccgtcg acctctagct agagcttggc
     1681 gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa
     1741 catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac
     1801 attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca
     1861 ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc
     1921 ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc
     1981 aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc
     2041 aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag
     2101 gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc
     2161 gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt
     2221 tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct
     2281 ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg
     2341 ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct
     2401 tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat
     2461 tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg
     2521 ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa
     2581 aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt
     2641 ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc
     2701 tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt
     2761 atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta
     2821 aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat
     2881 ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac
     2941 tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg
     3001 ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag
     3061 tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt
     3121 aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt
     3181 gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt
     3241 tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt
     3301 cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct
     3361 tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt
     3421 ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac
     3481 cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa
     3541 actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa
     3601 ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca
     3661 aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct
     3721 ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga
     3781 atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc
     3841 tgacgtcgac ggatcgggag atctcccgat cccctatggt gcactctcag tacaatctgc
     3901 tctgatgccg catagttaag ccagtatctg ctccctgctt gtgtgttgga ggtcgctgag
     3961 tagtgcgcga gcaaaattta agctacaaca aggcaaggct tgaccgacaa ttgcatgaag
     4021 aatctgctta gggttaggcg ttttgcgctg cttcgcgatg tacgggccag atatacgcgt
     4081 tgacattgat tattgactag ttattaatag taatcaatta cggggtcatt agttcatagc
     4141 ccatatatgg agttccgcgt tacataactt acggtaaatg gcccgcctgg ctgaccgccc
     4201 aacgaccccc gcccattgac gtcaataatg acgtatgttc ccatagtaac gccaataggg
     4261 actttccatt gacgtcaatg ggtggagtat ttacggtaaa ctgcccactt ggcagtacat
     4321 caagtgtatc atatgccaag tacgccccct attgacgtca atgacggtaa atggcccgcc
     4381 tggcattatg cccagtacat gaccttatgg gactttccta cttggcagta catctacgta
     4441 ttagtcatcg ctattaccat ggtgatgcgg ttttggcagt acatcaatgg gcgtggatag
     4501 cggtttgact cacggggatt tccaagtctc caccccattg acgtcaatgg gagtttgttt
     4561 tggcaccaaa atcaacggga ctttccaaaa tgtcgtaaca actccgcccc attgacgcaa
     4621 atgggcggta ggcgtgtacg gtgggaggtc tatataagca gagctctctg gctaactaga
     4681 gaacccactg cttactggct tatcgaaatt aatacgactc actataggga gacccaagct
     4741 ggctagcgtt taaacttaag atggaagccc ctgaatacct tgatttggat gaaattgact
     4801 ttagtgatga catatcttat tcagtcacat cactcaagac gatcccagaa ctgtgccgaa
     4861 gatgtgatac gcaaaacgaa gacagatcag tttctagctc tagctggaat tgtggcatct
     4921 caactcttat tacaaacacg caaaagccca caggaatcgc tgatgtgtac agtaagttcc
     4981 gcccagtgaa gcgggtttcg ccactgaaac atcagccaga gactctggag aacaatgaaa
     5041 gtgatgacca aaagaaccag aaagtggttg agtaccagaa agggggtgag tctgacctgg
     5101 gcccccagcc tcaggagctt ggccctggag atggagtggg cggcccacca ggtaagagct
     5161 ctgagcccag cacatcgctg ggtgaactgg agcactacga cctcgacatg gatgagattc
     5221 tggatgtgcc ttatattaaa tccagtcagc agcttgcctc ttttaccaag gtgacttcag
     5281 aaaaaagaat tttgggctta tgcacaacca tcaatggcct ttctggcaaa gcctgctcta
     5341 caggaagttc tgagagctca tcatccaaca tggcaccatt ttgtgttctt tctcccgtga
     5401 aaagccctca cttgagaaaa gcatcagctg tcatccacga ccagcacaag ctgtccactg
     5461 aagaaaccga gatctcacct cctctggtta aatgtggctc tgcatatgag cctgaaaacc
     5521 agagtaaaga cttcctaaac aagacattta gtgatcctca tggtcgaaaa gttgagaaga
     5581 caacaccaga ctgccagctc agggccttcc acctacaatc ctcagcagca gaatccaaac
     5641 cagaagagca ggtcagtggc ctaaaccgga ccagctccca aggcccagaa gaaaggagtg
     5701 agtatctgaa aaaagtgaaa agcatcttga acattgttaa agaaggacag atctctctcc
     5761 tgccacacct agctgcagac aatctagaca aaattcacga cgaaaatgga aacaatctat
     5821 tacatattgc ggcgtcacag ggacacgcag agtgtctaca gcacctcact tctttgatgg
     5881 gagaagactg cctcaatgag cgcaacactg agaagttgac tccagcaggc ctggccatta
     5941 agaatggtca gttggagtgc gtacgctgga tggtgagcga aacagaagcc attgcagaac
     6001 tgagttgttc taaggatttt ccaagcctta ttcattacgc aggttgctat ggccaggaaa
     6061 agattcttct gtggcttctt cagtttatgc aagaacaggg catctcgttg gatgaagtag
     6121 accaggatgg caacagtgcc gttcacgtag cctcacagca tggctacctt ggatgcatac
     6181 agaccttggt tgaatatgga gcaaatgtca ccatgcagaa ccacgctggg gaaaagccct
     6241 cccagagcgc cgagcggcag gggcacaccc tgtgctccag gtacctggtg gtggtggaga
     6301 cctgcatgtc gctggcctct caagtggtga agttaaccaa gcagctaaag gaacaaacag
     6361 tagaacgtgt cacgctgcag aaccaactcc aacaatttct agaagcccag aaatcagagg
     6421 gcaagtcact cccttcttca cccagttcgc catcctcacc tgcctccaga aagtcccagt
     6481 ggaaatctcc agatgcagat gatgattctg tagccaaaag caagccagga gtccaagagg
     6541 ggattcaggt tcttggaagc ctgtcagcct ccagccgggc tagacccaaa gcaaaagatg
     6601 aagattctga taaaatctta cgccagttat tgggaaagga aatctcagaa aatgtctgca
     6661 cccaggaaaa actgtccttg gaattccagg atgctcaggc ttcctctaga aattctaaaa
     6721 agatcccact ggagaagagg gaactgaagt tagccaggct gagacagctg atgcagaggt
     6781 cactgagtga gtctgacaca gactccaaca actctgagga ccccaagact accccagtga
     6841 ggaaggctga ccgaccaagg ccgcagccca ttgtagaaag cgtagagagt atggacagcg
     6901 cagaaagcct gcacctgatg attaagaaac acaccttggc atcaggggga cgcaggtttc
     6961 ctttcagcat caaggcctcc aaatccctgg atggccacag cccatctccc acctcagaga
     7021 gcagcgaacc agacttagaa tcccagtatc caggctcagg gagtattcct ccaaaccagc
     7081 cctctggtga ccctcagcag cccagccctg acagtactgc tgcccagaaa gttgccacaa
     7141 gtcccaagag tgccctcaag tctccatctt ccaagcgtag gacatctcag aacttaaaac
     7201 tgagagttac ctttgaggag cctgtggtgc agatggagca gcctagcctt gaactgaatg
     7261 gagaaaaaga caaagataag ggaaggactc tccagcggac ctccacaagt aacgaatcgg
     7321 gggatcaact gaaaaggcct tttggagcct ttcgatctat catggagaca ctaagtggca
     7381 accaaaacaa taataataac taccaggcag ccaaccagct gaaaacctct acattgccct
     7441 tgacctcact tgggaggaag acagatgcca agggaaaccc tgccagctcc gctagcaaag
     7501 gaaagaataa ggcagcactc gagaagaacg gcatcaaggc caacttcaag atccgccaca
     7561 acatcgagga cggcggcgtg cagctcgccg accactacca gcagaacacc cccatcggcg
     7621 acggccccgt gctgctgccc gacaaccact acctgagcta ccagtccgcc ctgagcaaag
     7681 accccaacga gaagcgcgat cacatggtcc tgctggagtt cgtgaccgcc gccgggatca
     7741 ctctcggcat ggacgagctg tacaagtaat ctagagggcc cgtttaaacc cgctgatcag
     7801 cctcgactgt gccttctagt tgccagccat ctgttgtttg cccctccccc gtgccttcct
     7861 tgaccctgga aggtgccact cccactgtcc tttcctaata aaatgaggaa attgcatcgc
     7921 attgtctgag taggtgtcat tctattctgg ggggtggggt ggggcaggac agcaaggggg
     7981 aggattggga agacaatagc aggcatgctg gggatgcggt gggctctatg gcttctgagg
     8041 cggaaagaac cagctggggc tctagggggt atccccacgc gccctgtagc ggcgcattaa
     8101 gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgccagc gccctagcgc
     8161 ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag
     8221 ctctaaatcg ggggctccct ttagggttcc gatttagtgc tttacggcac ctcgacccca
     8281 aaaaacttga ttagggtgat ggttcacgta gtgggccatc gccctgatag acggtttttc
     8341 gccctttgac gttggagtcc acg
//
