LOCUS       sequence-420307-        5211 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Mammalian expression of 3H-alpha-synuclein.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5211)
  AUTHORS   Wu S, Hernandez Villegas NC, Sirkis DW, Thomas-Wright I, 
            Wade-Martins R, Schekman R
  TITLE     Unconventional secretion of alpha-synuclein mediated by 
            palmitoylated DNAJC5 oligomers.
  JOURNAL   Elife. 2023 Jan 10;12:e85837. doi: 10.7554/eLife.85837.
  PUBMED    36626307
REFERENCE   2  (bases 1 to 5211)
  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: "Elife."; 
            date: "2023-01-10"; volume: "12:e85837. doi"; pages: " 
            10.7554/eLife.85837"
COMMENT     Sequence Label: 3H-alpha-synuclein
FEATURES             Location/Qualifiers
     source          1..5211
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..304
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        305..508
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     459..479
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     misc_feature    531..611
                     /label=MCS
                     /note="multiple cloning site"
     CDS             1344..1361
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    1939..2060
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1976..1995)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2030..2049
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2067..2522)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2204..2225)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2416..2435
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2549..2653
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2655..3012
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2655..2846
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2678..2698)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      2863..2998
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3047..3841
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             3047..3841
                     /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
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(3101..3120)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3711..3730
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4029..4048)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4073..4120
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4449..5037
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4938..4957
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
       61 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      121 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      181 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      241 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      301 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      361 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      421 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      481 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgcta
      541 ccggactcag atctcgagct caagcttcga attctgcagt cgacggtacc gcgggcccgg
      601 gatccaccgg tgccaccatg aacgaggatg acatgaaaaa gctgtataag caaatggtac
      661 aagaattgga gaaagccaga gacaggatgg aaaagcttta taaggagatg gtggagctca
      721 tacagaaagc gatcgagctc atgcggaaaa ttttccaaga ggtgaaacag gaggtggaaa
      781 aggccataga ggaaatgaaa aagctttatg acgaggcaaa aaagaaaatc gaacaaatga
      841 ttcagcaaat taaacaaggg ggagacaaac agaagatgga ggagttgctc aaaagagcaa
      901 aagaggaaat gaagaaggtt aaggacaaaa tggaaaagct gctggaaaag ctgaagcaga
      961 tcatgcaaga ggcaaagcag aagatggaaa agctcctcaa gcaactcaag gaggagatga
     1021 agaagatgaa agagaaaatg gaaaagcttc tcaaggaaat gaagcaacgc atggaagaag
     1081 tgaagaaaaa aatggacggc gatgatgaac tgcttgaaaa gatcaaaaag aatatcgacg
     1141 atctcaaaaa aatcgccgaa gatctgataa aaaaagctga ggaaaatata aaggaagcca
     1201 agaaaattgc tgaacagctg gtcaaacggg ctaagcaact tatcgaaaag gcgaagcagg
     1261 tcgcagaaga actcattaaa aaaattctgc agttgatcga gaaagccaaa gaaatcgcgg
     1321 agaaggtttt gaaaggactg gagcaccacc atcatcatca tgggggtagt gggggaggga
     1381 gcggtggcgg tagcggagat gtattcatga aaggactttc aaaggccaag gagggagttg
     1441 tggctgctgc tgagaaaacc aaacagggtg tggcagaagc agcaggaaag acaaaagagg
     1501 gtgttctcta tgtaggctcc aaaaccaagg agggagtggt gcatggtgtg gcaacagtgg
     1561 ctgagaagac caaagagcaa gtgacaaatg ttggaggagc agtggtgacg ggtgtgacag
     1621 cagtagccca gaagacagtg gagggagcag ggagcattgc agcagccact ggctttgtca
     1681 aaaaggacca gttgggcaag aatgaagaag gagccccaca ggaaggaatt ctggaagata
     1741 tgcctgtgga tcctgacaat gaggcttatg aaatgccttc tgaggaaggg tatcaagact
     1801 acgaacctga agcctaagcg gccgcgactc tagatcataa tcagccatac cacatttgta
     1861 gaggttttac ttgctttaaa aaacctccca cacctccccc tgaacctgaa acataaaatg
     1921 aatgcaattg ttgttgttaa cttgtttatt gcagcttata atggttacaa ataaagcaat
     1981 agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc
     2041 aaactcatca atgtatctta aggcgtaaat tgtaagcgtt aatattttgt taaaattcgc
     2101 gttaaatttt tgttaaatca gctcattttt taaccaatag gccgaaatcg gcaaaatccc
     2161 ttataaatca aaagaataga ccgagatagg gttgagtgtt gttccagttt ggaacaagag
     2221 tccactatta aagaacgtgg actccaacgt caaagggcga aaaaccgtct atcagggcga
     2281 tggcccacta cgtgaaccat caccctaatc aagttttttg gggtcgaggt gccgtaaagc
     2341 actaaatcgg aaccctaaag ggagcccccg atttagagct tgacggggaa agccggcgaa
     2401 cgtggcgaga aaggaaggga agaaagcgaa aggagcgggc gctagggcgc tggcaagtgt
     2461 agcggtcacg ctgcgcgtaa ccaccacacc cgccgcgctt aatgcgccgc tacagggcgc
     2521 gtcaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat
     2581 acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg
     2641 aaaaaggaag agtcctgagg cggaaagaac cagctgtgga atgtgtgtca gttagggtgt
     2701 ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca
     2761 gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat
     2821 ctcaattagt cagcaaccat agtcccgccc ctaactccgc ccatcccgcc cctaactccg
     2881 cccagttccg cccattctcc gccccatggc tgactaattt tttttattta tgcagaggcc
     2941 gaggccgcct cggcctctga gctattccag aagtagtgag gaggcttttt tggaggccta
     3001 ggcttttgca aagatcgatc aagagacagg atgaggatcg tttcgcatga ttgaacaaga
     3061 tggattgcac gcaggttctc cggccgcttg ggtggagagg ctattcggct atgactgggc
     3121 acaacagaca atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc
     3181 ggttcttttt gtcaagaccg acctgtccgg tgccctgaat gaactgcaag acgaggcagc
     3241 gcggctatcg tggctggcca cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac
     3301 tgaagcggga agggactggc tgctattggg cgaagtgccg gggcaggatc tcctgtcatc
     3361 tcaccttgct cctgccgaga aagtatccat catggctgat gcaatgcggc ggctgcatac
     3421 gcttgatccg gctacctgcc cattcgacca ccaagcgaaa catcgcatcg agcgagcacg
     3481 tactcggatg gaagccggtc ttgtcgatca ggatgatctg gacgaagagc atcaggggct
     3541 cgcgccagcc gaactgttcg ccaggctcaa ggcgagcatg cccgacggcg aggatctcgt
     3601 cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg
     3661 attcatcgac tgtggccggc tgggtgtggc ggaccgctat caggacatag cgttggctac
     3721 ccgtgatatt gctgaagagc ttggcggcga atgggctgac cgcttcctcg tgctttacgg
     3781 tatcgccgct cccgattcgc agcgcatcgc cttctatcgc cttcttgacg agttcttctg
     3841 agcgggactc tggggttcga aatgaccgac caagcgacgc ccaacctgcc atcacgagat
     3901 ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt ccgggacgcc
     3961 ggctggatga tcctccagcg cggggatctc atgctggagt tcttcgccca ccctaggggg
     4021 aggctaactg aaacacggaa ggagacaata ccggaaggaa cccgcgctat gacggcaata
     4081 aaaagacaga ataaaacgca cggtgttggg tcgtttgttc ataaacgcgg ggttcggtcc
     4141 cagggctggc actctgtcga taccccaccg agaccccatt ggggccaata cgcccgcgtt
     4201 tcttcctttt ccccacccca ccccccaagt tcgggtgaag gcccagggct cgcagccaac
     4261 gtcggggcgg caggccctgc catagcctca ggttactcat atatacttta gattgattta
     4321 aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc
     4381 aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa
     4441 ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca
     4501 ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta
     4561 actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc
     4621 caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca
     4681 gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta
     4741 ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag
     4801 cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt
     4861 cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc
     4921 acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac
     4981 ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac
     5041 gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc
     5101 tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgccatgca ttagttatta
     5161 atagtaatca attacggggt cattagttca tagcccatat atggagttcc g
//
