LOCUS       Exported                5152 bp ds-DNA     circular SYN 22-NOV-2023
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    EGFP-alphasynuclein-WT
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5152)
  AUTHORS   Furlong RA, Narain Y, Rankin J, Wyttenbach A, Rubinsztein DC
  TITLE     Alpha-synuclein overexpression promotes aggregation of mutant 
            huntingtin.
  JOURNAL   Biochem J. 2000 Mar 15;346 Pt 3:577-81.
  PUBMED    10698681
REFERENCE   2  (bases 1 to 5152)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Friday, Jan 19, 2024 from SnapGene 5.0.8
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..5152
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     enhancer        115..418
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        419..622
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     573..593
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     635..692
                     /label=Primer 1
     primer_bind     635..688
                     /label=Synuclein-Fw
     primer_bind     complement(651..666)
                     /label=Vector.REV
     primer_bind     complement(653..670)
                     /label=Vector.REV clover
     misc_feature    655
                     /label=cleave
     regulatory      661..666
                     /regulatory_class="other"
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     primer_bind     665..689
                     /label=CS353_2Asnca_F59.4
     CDS             667..1086
                     /codon_start=1
                     /label=SNCA
                     /translation="MDVFMKGLSKAKEGVVAAAEKTKQGVAEAAGKTKEGVLYVGSKTK
                     EGVVHGVTTVAEKTKEQVTNVGGAVVTGVTAVAQKTVEGAGSIAAATGFVKKDQLGKNE
                     EGAPQEGILEDMPVDPDNEAYEMPSEEGYQDYEPEA"
     primer_bind     complement(1066..1113)
                     /label=Synuclein-rev
     primer_bind     complement(1066..1113)
                     /label=CS354_SNCAgs_R63
     primer_bind     1087..1134
                     /label=FRET+con_Clover_fw
     primer_bind     1087..1132
                     /label=CT-fw
     misc_feature    1087..1113
                     /label=GS
     primer_bind     complement(1096..1113)
                     /label=CS156
     primer_bind     1112..1134
                     /label=mClover-FRET-NC-fw
     primer_bind     1112..1134
                     /label=mClover-FRET-NC-fw (P2A)
     primer_bind     1112..1128
                     /label=Fragment.FOR clover
     CDS             1114..1830
                     /codon_start=1
                     /product="bright green-yellow fluorescent protein derived 
                     from mClover2, with improved photostability (Bajar et al., 
                     2003)"
                     /label=mClover3
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGVACFSRYPDHMKQHDFFKSAMPEGYVQERTISFKDD
                     GTYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHYVYITADKQKNCIK
                     ANFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSHQSKLSKDPNEKRDHMVLL
                     EFVTAAGITHGMDELYK"
     primer_bind     complement(1159..1180)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     1767..1788
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1807..1861)
                     /label=CT-rev
     primer_bind     complement(1807..1860)
                     /label=FRET+con_Clover_rev
     primer_bind     complement(1807..1860)
                     /label=mClover-FRET-NC-rev
     primer_bind     complement(1807..1831)
                     /label=Fragment.REV
     misc_feature    1834
                     /label=cleave
     primer_bind     1948..1961
                     /label=CS156
     polyA_signal    1994..2115
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2031..2050)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2085..2104
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2122..2577)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2259..2280)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2471..2490
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2604..2708
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2710..3067
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     primer_bind     complement(2733..2753)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      2918..3053
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2980..2999
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     primer_bind     complement(3003..3014)
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3102..3896
                     /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(3156..3175)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3766..3785
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4084..4103)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4128..4175
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4504..5092
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4993..5012
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgccat gcattagtta
       61 ttaatagtaa tcaattacgg ggtcattagt tcatagccca tatatggagt tccgcgttac
      121 ataacttacg gtaaatggcc cgcctggctg accgcccaac gacccccgcc cattgacgtc
      181 aataatgacg tatgttccca tagtaacgcc aatagggact ttccattgac gtcaatgggt
      241 ggagtattta cggtaaactg cccacttggc agtacatcaa gtgtatcata tgccaagtac
      301 gccccctatt gacgtcaatg acggtaaatg gcccgcctgg cattatgccc agtacatgac
      361 cttatgggac tttcctactt ggcagtacat ctacgtatta gtcatcgcta ttaccatggt
      421 gatgcggttt tggcagtaca tcaatgggcg tggatagcgg tttgactcac ggggatttcc
      481 aagtctccac cccattgacg tcaatgggag tttgttttgg caccaaaatc aacgggactt
      541 tccaaaatgt cgtaacaact ccgccccatt gacgcaaatg ggcggtaggc gtgtacggtg
      601 ggaggtctat ataagcagag ctggtttagt gaaccgtcag atccgctagc gctaccggtc
      661 gccaccatgg atgtattcat gaaaggactt tcaaaggcca aggagggagt tgtggctgct
      721 gctgagaaaa ccaaacaggg tgtggcagaa gcagcaggaa agacaaaaga gggtgttctc
      781 tatgtaggct ccaaaaccaa ggagggagtg gtgcatggtg tgacaacagt ggctgagaag
      841 accaaagagc aagtgacaaa tgttggagga gcagtggtga cgggtgtgac agcagtagcc
      901 cagaagacag tggagggagc agggagcatt gcagcagcca ctggctttgt caaaaaggac
      961 cagttgggca agaatgaaga aggagcccca caggaaggaa ttctggaaga tatgcctgtg
     1021 gatcctgaca atgaggctta tgaaatgcct tctgaggaag ggtatcaaga ctacgaacct
     1081 gaagccggag gttcaggttc aggtggagga tccatggtga gcaagggcga ggagctgttc
     1141 accggggtgg tgcccatcct ggtcgagctg gacggcgacg taaacggcca caagttcagc
     1201 gtccgcggcg agggcgaggg cgatgccacc aacggcaagc tgaccctgaa gttcatctgc
     1261 accaccggca agctgcccgt gccctggccc accctcgtga ccaccttcgg ctacggcgtg
     1321 gcctgcttca gccgctaccc cgaccacatg aagcagcacg acttcttcaa gtccgccatg
     1381 cccgaaggct acgtccagga gcgcaccatc tctttcaagg acgacggtac ctacaagacc
     1441 cgcgccgagg tgaagttcga gggcgacacc ctggtgaacc gcatcgagct gaagggcatc
     1501 gacttcaagg aggacggcaa catcctgggg cacaagctgg agtacaactt caacagccac
     1561 tacgtctata tcacggccga caagcagaag aactgcatca aggctaactt caagatccgc
     1621 cacaacgttg aggacggcag cgtgcagctc gccgaccact accagcagaa cacccccatc
     1681 ggcgacggcc ccgtgctgct gcccgacaac cactacctga gccatcagtc caagctgagc
     1741 aaagacccca acgagaagcg cgatcacatg gtcctgctgg agttcgtgac cgccgccggg
     1801 attacacatg gcatggacga gctgtacaag taagaattct gcagtcgacg gtaccgcggg
     1861 cccgggatcc accggatcta gataactgat cataatcagc cataccacat ttgtagaggt
     1921 tttacttgct ttaaaaaacc tcccacacct ccccctgaac ctgaaacata aaatgaatgc
     1981 aattgttgtt gttaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat
     2041 cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact
     2101 catcaatgta tcttaacgcg taaattgtaa gcgttaatat tttgttaaaa ttcgcgttaa
     2161 atttttgtta aatcagctca ttttttaacc aataggccga aatcggcaaa atcccttata
     2221 aatcaaaaga atagaccgag atagggttga gtgttgttcc agtttggaac aagagtccac
     2281 tattaaagaa cgtggactcc aacgtcaaag ggcgaaaaac cgtctatcag ggcgatggcc
     2341 cactacgtga accatcaccc taatcaagtt ttttggggtc gaggtgccgt aaagcactaa
     2401 atcggaaccc taaagggagc ccccgattta gagcttgacg gggaaagccg gcgaacgtgg
     2461 cgagaaagga agggaagaaa gcgaaaggag cgggcgctag ggcgctggca agtgtagcgg
     2521 tcacgctgcg cgtaaccacc acacccgccg cgcttaatgc gccgctacag ggcgcgtcag
     2581 gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt
     2641 caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa
     2701 ggaagagtcc tgaggcggaa agaaccagct gtggaatgtg tgtcagttag ggtgtggaaa
     2761 gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac
     2821 caggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa
     2881 ttagtcagca accatagtcc cgcccctaac tccgcccatc ccgcccctaa ctccgcccag
     2941 ttccgcccat tctccgcccc atggctgact aatttttttt atttatgcag aggccgaggc
     3001 cgcctcggcc tctgagctat tccagaagta gtgaggaggc ttttttggag gcctaggctt
     3061 ttgcaaagat cgatcaagag acaggatgag gatcgtttcg catgattgaa caagatggat
     3121 tgcacgcagg ttctccggcc gcttgggtgg agaggctatt cggctatgac tgggcacaac
     3181 agacaatcgg ctgctctgat gccgccgtgt tccggctgtc agcgcagggg cgcccggttc
     3241 tttttgtcaa gaccgacctg tccggtgccc tgaatgaact gcaagacgag gcagcgcggc
     3301 tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag
     3361 cgggaaggga ctggctgcta ttgggcgaag tgccggggca ggatctcctg tcatctcacc
     3421 ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat gcggcggctg catacgcttg
     3481 atccggctac ctgcccattc gaccaccaag cgaaacatcg catcgagcga gcacgtactc
     3541 ggatggaagc cggtcttgtc gatcaggatg atctggacga agagcatcag gggctcgcgc
     3601 cagccgaact gttcgccagg ctcaaggcga gcatgcccga cggcgaggat ctcgtcgtga
     3661 cccatggcga tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt tctggattca
     3721 tcgactgtgg ccggctgggt gtggcggacc gctatcagga catagcgttg gctacccgtg
     3781 atattgctga agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg
     3841 ccgctcccga ttcgcagcgc atcgccttct atcgccttct tgacgagttc ttctgagcgg
     3901 gactctgggg ttcgaaatga ccgaccaagc gacgcccaac ctgccatcac gagatttcga
     3961 ttccaccgcc gccttctatg aaaggttggg cttcggaatc gttttccggg acgccggctg
     4021 gatgatcctc cagcgcgggg atctcatgct ggagttcttc gcccacccta gggggaggct
     4081 aactgaaaca cggaaggaga caataccgga aggaacccgc gctatgacgg caataaaaag
     4141 acagaataaa acgcacggtg ttgggtcgtt tgttcataaa cgcggggttc ggtcccaggg
     4201 ctggcactct gtcgataccc caccgagacc ccattggggc caatacgccc gcgtttcttc
     4261 cttttcccca ccccaccccc caagttcggg tgaaggccca gggctcgcag ccaacgtcgg
     4321 ggcggcaggc cctgccatag cctcaggtta ctcatatata ctttagattg atttaaaact
     4381 tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat
     4441 cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc
     4501 ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct
     4561 accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg
     4621 cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt taggccacca
     4681 cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc
     4741 tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga
     4801 taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac
     4861 gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga
     4921 agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag
     4981 ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg
     5041 acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag
     5101 caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca tg
//
