LOCUS       sequence-222076-        7128 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7128)
  AUTHORS   Cummings CJ, Mancini MA, Antalffy B, DeFranco DB, Orr HT, Zoghbi HY
  TITLE     Chaperone suppression of aggregation and altered subcellular 
            proteasome localization imply protein misfolding in SCA1.
  JOURNAL   Nat Genet. 1998 Jun;19(2):148-54.
  PUBMED    9620770
REFERENCE   2  (bases 1 to 7128)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nat 
            Genet."; date: "1998-06"; volume: "19(2)"; pages: "148-54"
COMMENT     Sequence Label: pEGFP-ATXN1-2Q
FEATURES             Location/Qualifiers
     source          1..7128
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        61..364
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        365..568
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     519..539
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      607..616
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(613..615,616..618,619..1329)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(613..615,616..618,619..1329)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(658..679)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(919..938)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1266..1287
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             3706..3717
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(3744..3760)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3745..3761)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     polyA_signal    3916..4037
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3953..3972)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4007..4026
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(4044..4499)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4181..4202)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4393..4412
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4526..4630
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4632..4989
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4632..4823
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4655..4675)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4840..4975
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             5024..5818
                     /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             5024..5818
                     /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(5078..5097)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5688..5707
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(6006..6025)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    6050..6097
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6426..7014
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6915..6934
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg
       61 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
      121 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      181 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      241 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      301 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      361 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      421 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      481 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      541 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgcta
      601 ccggtcgcca ccatggtgag caagggcgag gagctgttca ccggggtggt gcccatcctg
      661 gtcgagctgg acggcgacgt aaacggccac aagttcagcg tgtccggcga gggcgagggc
      721 gatgccacct acggcaagct gaccctgaag ttcatctgca ccaccggcaa gctgcccgtg
      781 ccctggccca ccctcgtgac caccctgacc tacggcgtgc agtgcttcag ccgctacccc
      841 gaccacatga agcagcacga cttcttcaag tccgccatgc ccgaaggcta cgtccaggag
      901 cgcaccatct tcttcaagga cgacggcaac tacaagaccc gcgccgaggt gaagttcgag
      961 ggcgacaccc tggtgaaccg catcgagctg aagggcatcg acttcaagga ggacggcaac
     1021 atcctggggc acaagctgga gtacaactac aacagccaca acgtctatat catggccgac
     1081 aagcagaaga acggcatcaa ggtgaacttc aagatccgcc acaacatcga ggacggcagc
     1141 gtgcagctcg ccgaccacta ccagcagaac acccccatcg gcgacggccc cgtgctgctg
     1201 cccgacaacc actacctgag cacccagtcc gccctgagca aagaccccaa cgagaagcgc
     1261 gatcacatgg tcctgctgga gttcgtgacc gccgccggga tcactctcgg catggacgag
     1321 ctgtacaagt ccggactcag atctcgagct caagctagct tcgaattcat gaaatccaac
     1381 caagagcgga gcaacgaatg cctgcctccc aagaagcgcg agatccccgc caccagccgg
     1441 tcctccgagg agaaggcccc taccctgccc agcgacaacc accgggtgga gggcacagca
     1501 tggctcccgg gcaaccctgg tggccggggc cacgggggcg ggaggcatgg gccggcaggg
     1561 acctcggtgg agcttggttt acaacaggga ataggtttac acaaagcatt gtccacaggg
     1621 ctggactact ccccgcccag cgctcccagg tctgtccccg tggccaccac gctgcctgcc
     1681 gcgtacgcca ccccgcagcc agggaccccg gtgtcccccg tacagtacgc tcacctgccg
     1741 cacaccttcc agttcattgg gtcctcccaa tacagtggaa cctatgccag cttcatccca
     1801 tcacagctga tccccccaac cgccaacccc gtcaccagtg cagtggcctc ggccgcaggg
     1861 gccaccactc catcccagcg ctcccagctg gaggcctatt ccactctgct ggccaacatg
     1921 ggcagtctga gccagacgcc gggacacaag gctgagcagc agcacctcag cagggctccg
     1981 gggcttatca ccccggggtc ccccccacca gcccagcaga accagtacgt ccacatttcc
     2041 agttctccgc agaacaccgg ccgcaccgcc tctcctccgg ccatccccgt ccacctccac
     2101 ccccaccaga cgatgatccc acacacgctc accctggggc ccccctccca ggtcgtcatg
     2161 caatacgccg actccggcag ccactttgtc cctcgggagg ccaccaagaa agctgagagc
     2221 agccggctgc agcaggccat ccaggccaag gaggtcctga acggtgagat ggagaagagc
     2281 cggcggtacg gggccccgtc ctcagccgac ctgggcctgg gcaaggcagg cggcaagtcg
     2341 gttcctcacc cgtacgagtc caggcacgtg gtggtccacc cgagcccctc agactacagc
     2401 agtcgtgatc cttcgggggt ccgggcctct gtgatggtcc tgcccaacag caacacgccc
     2461 gcagctgacc tggaggtgca acaggccact catcgtgaag cctccccttc taccctcaac
     2521 gacaaaagtg gcctgcattt agggaagcct ggccaccggt cctacgcgct ctcaccccac
     2581 acggtcattc agaccacaca cagtgcttca gagccactcc cggtgggact gccagccacg
     2641 gccttctacg cagggactca accccctgtc atcggctacc tgagcggcca gcagcaagca
     2701 atcacctacg ccggcagcct gccccagcac ctggtgatcc ccggcacaca gcccctgctc
     2761 atcccggtcg gcagcactga catggaagcg tcgggggcag ccccggccat agtcacgtca
     2821 tccccccagt ttgctgcagt gcctcacacg ttcgtcacca ccgcccttcc caagagcgag
     2881 aacttcaacc ctgaggccct ggtcacccag gccgcctacc cagccatggt gcaggcccag
     2941 atccacctgc ctgtggtgca gtccgtggcc tccccggcgg cggctccccc tacgctgcct
     3001 ccctacttca tgaaaggctc catcatccag ttggccaacg gggagctaaa gaaggtggaa
     3061 gacttaaaaa cagaagattt catccagagt gcagagataa gcaacgacct gaagatcgac
     3121 tccagcaccg tagagaggat tgaagacagc catagcccgg gcgtggccgt gatacagttc
     3181 gccgtcgggg agcaccgagc ccaggtcagc gttgaagttt tggtagagta tccttttttt
     3241 gtgtttggac agggctggtc atcctgctgt ccggagagaa ccagccagct ctttgatttg
     3301 ccgtgttcca aactctcagt tggggatgtc tgcatctcgc ttaccctcaa gaacctgaag
     3361 aacggctctg ttaaaaaggg ccagcccgtg gatcccgcca gcgtcctgct gaagcactca
     3421 aaggccgacg gcctggcggg cagcagacac aggtatgccg agcaggaaaa cggaatcaac
     3481 caggggagtg cccagatgct ctctgagaat ggcgaactga agtttccaga gaaaatggga
     3541 ttgcctgcag cgcccttcct caccaaaata gaacccagca agcccgcggc aacgaggaag
     3601 aggaggtggt cggcgccaga gagccgcaaa ctggagaagt cagaagacga accacctttg
     3661 actcttccta agccttctct aattcctcag gaggttaaga tttgcattga aggccggtct
     3721 aatgtaggca agtagaagct tatcgatacc gtcgacctcg agggggggcc cggtaccgcg
     3781 ggcccgggat ccaccggatc tagataactg atcataatca gccataccac atttgtagag
     3841 gttttacttg ctttaaaaaa cctcccacac ctccccctga acctgaaaca taaaatgaat
     3901 gcaattgttg ttgttaactt gtttattgca gcttataatg gttacaaata aagcaatagc
     3961 atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa
     4021 ctcatcaatg tatcttaacg cgtaaattgt aagcgttaat attttgttaa aattcgcgtt
     4081 aaatttttgt taaatcagct cattttttaa ccaataggcc gaaatcggca aaatccctta
     4141 taaatcaaaa gaatagaccg agatagggtt gagtgttgtt ccagtttgga acaagagtcc
     4201 actattaaag aacgtggact ccaacgtcaa agggcgaaaa accgtctatc agggcgatgg
     4261 cccactacgt gaaccatcac cctaatcaag ttttttgggg tcgaggtgcc gtaaagcact
     4321 aaatcggaac cctaaaggga gcccccgatt tagagcttga cggggaaagc cggcgaacgt
     4381 ggcgagaaag gaagggaaga aagcgaaagg agcgggcgct agggcgctgg caagtgtagc
     4441 ggtcacgctg cgcgtaacca ccacacccgc cgcgcttaat gcgccgctac agggcgcgtc
     4501 aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca
     4561 ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa
     4621 aaggaagagt cctgaggcgg aaagaaccag ctgtggaatg tgtgtcagtt agggtgtgga
     4681 aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca
     4741 accaggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc
     4801 aattagtcag caaccatagt cccgccccta actccgccca tcccgcccct aactccgccc
     4861 agttccgccc attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag
     4921 gccgcctcgg cctctgagct attccagaag tagtgaggag gcttttttgg aggcctaggc
     4981 ttttgcaaag atcgatcaag agacaggatg aggatcgttt cgcatgattg aacaagatgg
     5041 attgcacgca ggttctccgg ccgcttgggt ggagaggcta ttcggctatg actgggcaca
     5101 acagacaatc ggctgctctg atgccgccgt gttccggctg tcagcgcagg ggcgcccggt
     5161 tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa ctgcaagacg aggcagcgcg
     5221 gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct gtgctcgacg ttgtcactga
     5281 agcgggaagg gactggctgc tattgggcga agtgccgggg caggatctcc tgtcatctca
     5341 ccttgctcct gccgagaaag tatccatcat ggctgatgca atgcggcggc tgcatacgct
     5401 tgatccggct acctgcccat tcgaccacca agcgaaacat cgcatcgagc gagcacgtac
     5461 tcggatggaa gccggtcttg tcgatcagga tgatctggac gaagagcatc aggggctcgc
     5521 gccagccgaa ctgttcgcca ggctcaaggc gagcatgccc gacggcgagg atctcgtcgt
     5581 gacccatggc gatgcctgct tgccgaatat catggtggaa aatggccgct tttctggatt
     5641 catcgactgt ggccggctgg gtgtggcgga ccgctatcag gacatagcgt tggctacccg
     5701 tgatattgct gaagagcttg gcggcgaatg ggctgaccgc ttcctcgtgc tttacggtat
     5761 cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt cttgacgagt tcttctgagc
     5821 gggactctgg ggttcgaaat gaccgaccaa gcgacgccca acctgccatc acgagatttc
     5881 gattccaccg ccgccttcta tgaaaggttg ggcttcggaa tcgttttccg ggacgccggc
     5941 tggatgatcc tccagcgcgg ggatctcatg ctggagttct tcgcccaccc tagggggagg
     6001 ctaactgaaa cacggaagga gacaataccg gaaggaaccc gcgctatgac ggcaataaaa
     6061 agacagaata aaacgcacgg tgttgggtcg tttgttcata aacgcggggt tcggtcccag
     6121 ggctggcact ctgtcgatac cccaccgaga ccccattggg gccaatacgc ccgcgtttct
     6181 tccttttccc caccccaccc cccaagttcg ggtgaaggcc cagggctcgc agccaacgtc
     6241 ggggcggcag gccctgccat agcctcaggt tactcatata tactttagat tgatttaaaa
     6301 cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa
     6361 atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga
     6421 tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg
     6481 ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact
     6541 ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac
     6601 cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg
     6661 gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg
     6721 gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga
     6781 acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc
     6841 gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg
     6901 agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc
     6961 tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc
     7021 agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt
     7081 cctgcgttat cccctgattc tgtggataac cgtattaccg ccatgcat
//
