LOCUS       sequence-202606-        5836 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 5836)
  AUTHORS   Chai Y, Shao J, Miller VM, Williams A, Paulson HL
  TITLE     Live-cell imaging reveals divergent intracellular dynamics of 
            polyglutamine disease proteins and supports a sequestration model of
            pathogenesis.
  JOURNAL   Proc Natl Acad Sci U S A. 2002 Jul 9. 99(14):9310-5.
  PUBMED    12084819
REFERENCE   2  (bases 1 to 5836)
  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: "Proc Natl 
            Acad Sci U S A. 2002 Jul 9. 99(14):9310-5."
COMMENT     Sequence Label: pEGFP-C1-Ataxin3Q28
FEATURES             Location/Qualifiers
     source          1..5836
                     /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"
     misc_feature    1330..1395
                     /label=MCS
                     /note="multiple cloning site"
     polyA_signal    2624..2745
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2661..2680)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2715..2734
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2752..3207)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2889..2910)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3101..3120
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3234..3338
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3340..3697
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3340..3531
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3363..3383)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3548..3683
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3732..4526
                     /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             3732..4526
                     /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(3786..3805)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4396..4415
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4714..4733)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4758..4805
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5134..5722
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5623..5642
                     /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 caagcttcga attctgcagt cgacggtacc
     1381 gcgggcccgg gatccataaa catggagtcc atcttccacg agaaacaaga aggctcactt
     1441 tgtgctcaac attgcctgaa taacttattg caaggagaat attttagccc tgtggaatta
     1501 tcctcaattg cacatcagct ggatgaggag gagaggatga gaatggcaga aggaggagtt
     1561 actagtgaag attatcgcac gtttttacag cagccttctg gaaatatgga tgacagtggt
     1621 tttttctcta ttcaggttat aagcaatgcc ttgaaagttt ggggtttaga actaatcctg
     1681 ttcaacagtc cagagtatca gaggctcagg atcgatccta taaatgaaag atcatttata
     1741 tgcaattata aggaacactg gtttacagtt agaaaattag gaaaacagtg gtttaacttg
     1801 aattctctct tgacgggtcc agaattaata tcagatacat atcttgcact tttcttggct
     1861 caattacaac aggaaggtta ttctatattt gttgttaagg gtgatctgcc agattgcgaa
     1921 gctgaccaac tcctgcagat gattagggtc caacagatgc atcgaccaaa acttattgga
     1981 gaagaattag cacaactaaa agagcaaaga gtccataaaa cagacctgga acgaatgtta
     2041 gaagcaaatg atggctcagg aatgttagac gaagatgagg aggatttgca gagggctctg
     2101 gcactaagtc gccaagaaat tgacatggaa gatgaggaag cagatctccg cagggctatt
     2161 cagctaagta tgcaaggtag ttccagaaac atatctcaag atatgacaca gacatcaggt
     2221 acaaatctta cttcagaaga gcttcggaag agacgggaag cctactttga aaaacagcag
     2281 caaaagcagc aacagcagca gcagcagcag cagcagcagc agcagcagca gcagcagcag
     2341 cagcagcagc agcagcaggg ggacctatca ggacagagtt cacatccatg tgaaaggcca
     2401 gccaccagtt caggagcact tgggagtgat ctaggtaagg cctgctcacc attcatcatg
     2461 ttcgctacct tcacacttta tctgacataa gagcggatcc accggatcta gataactgat
     2521 cataatcagc cataccacat ttgtagaggt tttacttgct ttaaaaaacc tcccacacct
     2581 ccccctgaac ctgaaacata aaatgaatgc aattgttgtt gttaacttgt ttattgcagc
     2641 ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc
     2701 actgcattct agttgtggtt tgtccaaact catcaatgta tcttaacgcg taaattgtaa
     2761 gcgttaatat tttgttaaaa ttcgcgttaa atttttgtta aatcagctca ttttttaacc
     2821 aataggccga aatcggcaaa atcccttata aatcaaaaga atagaccgag atagggttga
     2881 gtgttgttcc agtttggaac aagagtccac tattaaagaa cgtggactcc aacgtcaaag
     2941 ggcgaaaaac cgtctatcag ggcgatggcc cactacgtga accatcaccc taatcaagtt
     3001 ttttggggtc gaggtgccgt aaagcactaa atcggaaccc taaagggagc ccccgattta
     3061 gagcttgacg gggaaagccg gcgaacgtgg cgagaaagga agggaagaaa gcgaaaggag
     3121 cgggcgctag ggcgctggca agtgtagcgg tcacgctgcg cgtaaccacc acacccgccg
     3181 cgcttaatgc gccgctacag ggcgcgtcag gtggcacttt tcggggaaat gtgcgcggaa
     3241 cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac
     3301 cctgataaat gcttcaataa tattgaaaaa ggaagagtcc tgaggcggaa agaaccagct
     3361 gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc tccccagcag gcagaagtat
     3421 gcaaagcatg catctcaatt agtcagcaac caggtgtgga aagtccccag gctccccagc
     3481 aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accatagtcc cgcccctaac
     3541 tccgcccatc ccgcccctaa ctccgcccag ttccgcccat tctccgcccc atggctgact
     3601 aatttttttt atttatgcag aggccgaggc cgcctcggcc tctgagctat tccagaagta
     3661 gtgaggaggc ttttttggag gcctaggctt ttgcaaagat cgatcaagag acaggatgag
     3721 gatcgtttcg catgattgaa caagatggat tgcacgcagg ttctccggcc gcttgggtgg
     3781 agaggctatt cggctatgac tgggcacaac agacaatcgg ctgctctgat gccgccgtgt
     3841 tccggctgtc agcgcagggg cgcccggttc tttttgtcaa gaccgacctg tccggtgccc
     3901 tgaatgaact gcaagacgag gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt
     3961 gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga ctggctgcta ttgggcgaag
     4021 tgccggggca ggatctcctg tcatctcacc ttgctcctgc cgagaaagta tccatcatgg
     4081 ctgatgcaat gcggcggctg catacgcttg atccggctac ctgcccattc gaccaccaag
     4141 cgaaacatcg catcgagcga gcacgtactc ggatggaagc cggtcttgtc gatcaggatg
     4201 atctggacga agagcatcag gggctcgcgc cagccgaact gttcgccagg ctcaaggcga
     4261 gcatgcccga cggcgaggat ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca
     4321 tggtggaaaa tggccgcttt tctggattca tcgactgtgg ccggctgggt gtggcggacc
     4381 gctatcagga catagcgttg gctacccgtg atattgctga agagcttggc ggcgaatggg
     4441 ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc atcgccttct
     4501 atcgccttct tgacgagttc ttctgagcgg gactctgggg ttcgaaatga ccgaccaagc
     4561 gacgcccaac ctgccatcac gagatttcga ttccaccgcc gccttctatg aaaggttggg
     4621 cttcggaatc gttttccggg acgccggctg gatgatcctc cagcgcgggg atctcatgct
     4681 ggagttcttc gcccacccta gggggaggct aactgaaaca cggaaggaga caataccgga
     4741 aggaacccgc gctatgacgg caataaaaag acagaataaa acgcacggtg ttgggtcgtt
     4801 tgttcataaa cgcggggttc ggtcccaggg ctggcactct gtcgataccc caccgagacc
     4861 ccattggggc caatacgccc gcgtttcttc cttttcccca ccccaccccc caagttcggg
     4921 tgaaggccca gggctcgcag ccaacgtcgg ggcggcaggc cctgccatag cctcaggtta
     4981 ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa
     5041 gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc
     5101 gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat
     5161 ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga
     5221 gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt
     5281 tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata
     5341 cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac
     5401 cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg
     5461 ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg
     5521 tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag
     5581 cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct
     5641 ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc
     5701 aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt
     5761 ttgctggcct tttgctcaca tgttctttcc tgcgttatcc cctgattctg tggataaccg
     5821 tattaccgcc atgcat
//
