LOCUS       Exported                5834 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  Overexpression of  synuclein alpha in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    pcDNA3.1-hSNCA-NE
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5834)
  AUTHORS   Ho PW, Leung CT, Liu H, Pang SY, Lam CS, Xian J, Li L, Kung MH, 
            Ramsden DB, Ho SL
  TITLE     Age-dependent accumulation of oligomeric SNCA/alpha-synuclein from 
            impaired degradation in mutant LRRK2 knockin mouse model of 
            Parkinson disease: role for therapeutic activation of 
            chaperone-mediated autophagy (CMA).
  JOURNAL   Autophagy. 2019 Apr 14:1-24. doi: 10.1080/15548627.2019.1603545.
  PUBMED    30983487
REFERENCE   2  (bases 1 to 5834)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..5834
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     enhancer        1..380
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        381..584
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     535..555
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     629..648
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        629..647
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1191..1208)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1197..1421
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      1467..1895
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1554..1573)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1764..1785
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(1904..1924)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        1909..2238
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2089..2224
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2151..2170
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             2305..3099
                     /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(2359..2378)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     2969..2988
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    3273..3394
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3310..3329)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3364..3383
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3443..3459)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3443..3459)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3456..3478)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3467..3483
                     /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(3491..3521)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3536..3557
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3674..3691)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3845..4433)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3925..3944)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(4604..5464)
                     /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     5227..5246
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5465..5569)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(5644..5663)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc
       61 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
      121 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
      181 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
      241 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
      301 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat
      361 tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc
      421 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt
      481 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa
      541 tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctctctgg ctaactagag
      601 aacccactgc ttactggctt atcgaaatta atacgactca ctatagggag acccaagctg
      661 gctagcgttt aaacttaagc ttatggatgt attcatgaaa ggactttcaa aggccaagga
      721 gggagttgtg gctgctgctg agaaaaccaa acagggtgtg gcagaagcag caggaaagac
      781 aaaagagggt gttctctatg taggctccaa aaccaaggag ggagtggtgc atggtgtggc
      841 aacagtggct gagaagacca aagagcaagt gacaaatgtt ggaggagcag tggtgacggg
      901 tgtgacagca gtagcccaga agacagtgga gggagcaggg agcattgcag cagccactgg
      961 ctttgtcaaa aaggaccagt tgggcaagaa tgaagaagga gccccacagg aaggaattct
     1021 ggaagatatg cctgtggatc ctgacaatga ggcttatgaa atgccttctg aggaagggta
     1081 tcaagactac gaacctgaag ccaccaaaga aaacccgcgt agcaaccagg aagaaagcta
     1141 tgatgataac gaaagctaat ctagagggcc cgtttaaacc cgctgatcag cctcgactgt
     1201 gccttctagt tgccagccat ctattgtttg cccctccccc gtgccttcct tgaccctgga
     1261 aggtgccact cccactgtcc tttcctaata aaatgaggaa attgcatcgc attgtctgag
     1321 taggtgtcat tctattctgg ggggtggggt ggggcaggac agcaaggggg aggattggga
     1381 agacaatagc aggcatgctg gggatgcggt gggctctatg gcttctgagg cggaaagaac
     1441 cagctggggc tctagggggt atccccacgc gccctgtagc ggcgcattaa gcgcggcggg
     1501 tgtggtggtt acgcgcagcg tgaccgctac acttgccagc gccctagcgc ccgctccttt
     1561 cgctttcttc ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg
     1621 ggggctccct ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga
     1681 ttagggtgat ggttcacgta gtgggccatc gccctgatag acggtttttc gccctttgac
     1741 gttggagtcc acgttcttta atagtggact cttgttccaa actggaacaa cactcaaccc
     1801 tatctcggtc tattcttttg atttataagg gattttgccg atttcggcct attggttaaa
     1861 aaatgagctg atttaacaaa aatttaacgc gaattaattc tgtggaatgt gtgtcagtta
     1921 gggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat
     1981 tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc
     2041 atgcatctca attagtcagc aaccatagtc ccgcccctaa ctccgcccat cccgccccta
     2101 actccgccca gttccgccca ttctccgccc catggctgac taattttttt tatttatgca
     2161 gaggccgagg ccgcctctgc ctctgagcta ttccagaagt agtgaggagg cttttttgga
     2221 ggcctaggct tttgcaaaaa gctcccggga gcttgtatat ccattttcgg atctgatcaa
     2281 gagacaggat gaggatcgtt tcgcatgatt gaacaagatg gattgcacgc aggttctccg
     2341 gccgcttggg tggagaggct attcggctat gactgggcac aacagacaat cggctgctct
     2401 gatgccgccg tgttccggct gtcagcgcag gggcgcccgg ttctttttgt caagaccgac
     2461 ctgtccggtg ccctgaatga actgcaggac gaggcagcgc ggctatcgtg gctggccacg
     2521 acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg aagcgggaag ggactggctg
     2581 ctattgggcg aagtgccggg gcaggatctc ctgtcatctc accttgctcc tgccgagaaa
     2641 gtatccatca tggctgatgc aatgcggcgg ctgcatacgc ttgatccggc tacctgccca
     2701 ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta ctcggatgga agccggtctt
     2761 gtcgatcagg atgatctgga cgaagagcat caggggctcg cgccagccga actgttcgcc
     2821 aggctcaagg cgcgcatgcc cgacggcgag gatctcgtcg tgacccatgg cgatgcctgc
     2881 ttgccgaata tcatggtgga aaatggccgc ttttctggat tcatcgactg tggccggctg
     2941 ggtgtggcgg accgctatca ggacatagcg ttggctaccc gtgatattgc tgaagagctt
     3001 ggcggcgaat gggctgaccg cttcctcgtg ctttacggta tcgccgctcc cgattcgcag
     3061 cgcatcgcct tctatcgcct tcttgacgag ttcttctgag cgggactctg gggttcgaaa
     3121 tgaccgacca agcgacgccc aacctgccat cacgagattt cgattccacc gccgccttct
     3181 atgaaaggtt gggcttcgga atcgttttcc gggacgccgg ctggatgatc ctccagcgcg
     3241 gggatctcat gctggagttc ttcgcccacc ccaacttgtt tattgcagct tataatggtt
     3301 acaaataaag caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta
     3361 gttgtggttt gtccaaactc atcaatgtat cttatcatgt ctgtataccg tcgacctcta
     3421 gctagagctt ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca
     3481 caattccaca caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag
     3541 tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt
     3601 cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc
     3661 gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg
     3721 tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa
     3781 agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg
     3841 cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga
     3901 ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg
     3961 tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg
     4021 gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc
     4081 gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg
     4141 gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca
     4201 ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt
     4261 ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag
     4321 ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg
     4381 gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc
     4441 ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt
     4501 tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt
     4561 ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca
     4621 gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg
     4681 tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac
     4741 cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg
     4801 ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc
     4861 gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta
     4921 caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac
     4981 gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc
     5041 ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac
     5101 tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact
     5161 caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa
     5221 tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt
     5281 cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca
     5341 ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa
     5401 aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac
     5461 tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg
     5521 gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc
     5581 gaaaagtgcc acctgacgtc gacggatcgg gagatctccc gatcccctat ggtgcactct
     5641 cagtacaatc tgctctgatg ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt
     5701 ggaggtcgct gagtagtgcg cgagcaaaat ttaagctaca acaaggcaag gcttgaccga
     5761 caattgcatg aagaatctgc ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc
     5821 cagatatacg cgtt
//
