LOCUS       Exported                5143 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 5143)
  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 5143)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Friday, Jan 19, 2024 from SnapGene 5.0.8
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..5143
                     /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
     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..1135
                     /label=CT-fw
     misc_feature    1087..1113
                     /label=GS
     primer_bind     complement(1096..1113)
                     /label=CS156
     primer_bind     1112..1135
                     /label=Fragment.FOR
     primer_bind     1112..1135
                     /label=FRET+con_Ruby_fw
     primer_bind     1112..1135
                     /label=mRuby-FRET-NC-fw
     CDS             1114..1821
                     /codon_start=1
                     /product="monomeric red fluorescent protein derived from 
                     mRuby2, with improved brightness and photostability (Bajar 
                     et al., 2016)"
                     /label=mRuby3
                     /translation="VSKGEELIKENMRMKVVMEGSVNGHQFKCTGEGEGRPYEGVQTMR
                     IKVIEGGPLPFAFDILATSFMYGSRTFIKYPADIPDFFKQSFPEGFTWERVTRYEDGGV
                     VTVTQDTSLEDGELVYNVKVRGVNFPSNGPVMQKKTKGWEPNTEMMYPADGGLRGYTDI
                     ALKVDGGGHLHCNFVTTYRSKKTVGNIKMPGVHAVDHRLERIEESDNETYVVQREVAVA
                     KYSNLGGGMDELYK"
     primer_bind     complement(1795..1852)
                     /label=CT-rev
     primer_bind     complement(1795..1822)
                     /label=Fragment.REV
     primer_bind     complement(1795..1821)
                     /label=FRET+con_Ruby_rev
     primer_bind     complement(1795..1821)
                     /label=mRuby-FRET-NC-rev
     primer_bind     complement(1795..1821)
                     /label=mRuby-FRET-NC-fw (P2A)
     misc_feature    1825
                     /label=cleave
     primer_bind     1939..1952
                     /label=CS156
     polyA_signal    1985..2106
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2022..2041)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2076..2095
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2113..2568)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2250..2271)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2462..2481
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2595..2699
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2701..3058
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     primer_bind     complement(2724..2744)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      2909..3044
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2971..2990
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     primer_bind     complement(2994..3005)
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3093..3887
                     /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(3147..3166)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3757..3776
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4075..4094)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4119..4166
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4495..5083
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4984..5003
                     /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 tccgtgtcta agggcgaaga gctgatcaag
     1141 gaaaatatgc gtatgaaggt ggtcatggaa ggttcggtca acggccacca attcaaatgc
     1201 acaggtgaag gagaaggcag accgtacgag ggagtgcaaa ccatgaggat caaagtcatc
     1261 gagggaggac ccctgccatt tgcctttgac attcttgcca cgtcgttcat gtatggcagc
     1321 cgtaccttta tcaagtaccc ggccgacatc cctgatttct ttaaacagtc ctttcctgag
     1381 ggttttactt gggaaagagt tacgagatac gaagatggtg gagtcgtcac cgtcacgcag
     1441 gacaccagcc ttgaggatgg cgagctcgtc tacaacgtca aggtcagagg ggtaaacttt
     1501 ccctccaatg gtcccgtgat gcagaagaag accaagggtt gggagcctaa tacagagatg
     1561 atgtatccag cagatggtgg tctgagagga tacactgaca tcgcactgaa agttgatggt
     1621 ggtggccatc tgcactgcaa cttcgtgaca acttacaggt caaaaaagac cgtcgggaac
     1681 atcaagatgc ccggtgtcca tgccgttgat caccgcctgg aaaggatcga ggagagtgac
     1741 aatgaaacct acgtagtgca aagagaagtg gcagttgcca aatacagcaa ccttggtggt
     1801 ggcatggacg agctgtacaa gtaagaattc tgcagtcgac ggtaccgcgg gcccgggatc
     1861 caccggatct agataactga tcataatcag ccataccaca tttgtagagg ttttacttgc
     1921 tttaaaaaac ctcccacacc tccccctgaa cctgaaacat aaaatgaatg caattgttgt
     1981 tgttaacttg tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt
     2041 cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt
     2101 atcttaacgc gtaaattgta agcgttaata ttttgttaaa attcgcgtta aatttttgtt
     2161 aaatcagctc attttttaac caataggccg aaatcggcaa aatcccttat aaatcaaaag
     2221 aatagaccga gatagggttg agtgttgttc cagtttggaa caagagtcca ctattaaaga
     2281 acgtggactc caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg
     2341 aaccatcacc ctaatcaagt tttttggggt cgaggtgccg taaagcacta aatcggaacc
     2401 ctaaagggag cccccgattt agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg
     2461 aagggaagaa agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc
     2521 gcgtaaccac cacacccgcc gcgcttaatg cgccgctaca gggcgcgtca ggtggcactt
     2581 ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt
     2641 atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagtc
     2701 ctgaggcgga aagaaccagc tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg
     2761 ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg
     2821 aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc
     2881 aaccatagtc ccgcccctaa ctccgcccat cccgccccta actccgccca gttccgccca
     2941 ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg ccgcctcggc
     3001 ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctaggct tttgcaaaga
     3061 tcgatcaaga gacaggatga ggatcgtttc gcatgattga acaagatgga ttgcacgcag
     3121 gttctccggc cgcttgggtg gagaggctat tcggctatga ctgggcacaa cagacaatcg
     3181 gctgctctga tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca
     3241 agaccgacct gtccggtgcc ctgaatgaac tgcaagacga ggcagcgcgg ctatcgtggc
     3301 tggccacgac gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg
     3361 actggctgct attgggcgaa gtgccggggc aggatctcct gtcatctcac cttgctcctg
     3421 ccgagaaagt atccatcatg gctgatgcaa tgcggcggct gcatacgctt gatccggcta
     3481 cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact cggatggaag
     3541 ccggtcttgt cgatcaggat gatctggacg aagagcatca ggggctcgcg ccagccgaac
     3601 tgttcgccag gctcaaggcg agcatgcccg acggcgagga tctcgtcgtg acccatggcg
     3661 atgcctgctt gccgaatatc atggtggaaa atggccgctt ttctggattc atcgactgtg
     3721 gccggctggg tgtggcggac cgctatcagg acatagcgtt ggctacccgt gatattgctg
     3781 aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc gccgctcccg
     3841 attcgcagcg catcgccttc tatcgccttc ttgacgagtt cttctgagcg ggactctggg
     3901 gttcgaaatg accgaccaag cgacgcccaa cctgccatca cgagatttcg attccaccgc
     3961 cgccttctat gaaaggttgg gcttcggaat cgttttccgg gacgccggct ggatgatcct
     4021 ccagcgcggg gatctcatgc tggagttctt cgcccaccct agggggaggc taactgaaac
     4081 acggaaggag acaataccgg aaggaacccg cgctatgacg gcaataaaaa gacagaataa
     4141 aacgcacggt gttgggtcgt ttgttcataa acgcggggtt cggtcccagg gctggcactc
     4201 tgtcgatacc ccaccgagac cccattgggg ccaatacgcc cgcgtttctt ccttttcccc
     4261 accccacccc ccaagttcgg gtgaaggccc agggctcgca gccaacgtcg gggcggcagg
     4321 ccctgccata gcctcaggtt actcatatat actttagatt gatttaaaac ttcattttta
     4381 atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg
     4441 tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga
     4501 tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt
     4561 ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag
     4621 agcgcagata ccaaatactg ttcttctagt gtagccgtag ttaggccacc acttcaagaa
     4681 ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag
     4741 tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca
     4801 gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac
     4861 cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa
     4921 ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc
     4981 agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg
     5041 tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc
     5101 ctttttacgg ttcctggcct tttgctggcc ttttgctcac atg
//
