LOCUS       sequence-353799-        5678 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  expresses mVenus and mCherry tagged to a mutant of the huntingtin 
            exon1 (polyQ54) in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5678)
  AUTHORS   Wan Q, Mouton SN, Veenhoff LM, Boersma AJ
  TITLE     A FRET-based method for monitoring structural transitions in protein
            self-organization.
  JOURNAL   Cell Rep Methods. 2022 Mar 28;2(3):100184. doi: 
            10.1016/j.crmeth.2022.100184. eCollection 2022 Mar 28.
  PUBMED    35475219
REFERENCE   2  (bases 1 to 5678)
  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: "Cell Rep 
            Methods."; date: "2022-03-28"; volume: "2(3):100184. doi"; pages: " 
            10.1016/j.crmeth.2022.100184. eCollection 2022 Mar 28"
COMMENT     Sequence Label: pQ54-VC
FEATURES             Location/Qualifiers
     source          1..5678
                     /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"
     CDS             join(983..985,986..1498)
                     /codon_start=1
                     /product="N-terminal fragment of mVenus for use in 
                     bimolecular fluorescence complementation (BiFC) (Kodama and
                     Hu, 2010)"
                     /label=VN173
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     LICTTGKLPVPWPTLVTTLGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIKA
                     NFKIRHNIE"
     primer_bind     complement(1025..1046)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1286..1305)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1633..1654
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1794..1812)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     2062..2081
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     polyA_signal    2466..2587
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2503..2522)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2557..2576
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2594..3049)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2731..2752)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2943..2962
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3076..3180
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3182..3539
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3182..3373
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3205..3225)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3390..3525
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3574..4368
                     /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             3574..4368
                     /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(3628..3647)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4238..4257
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4556..4575)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4600..4647
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4976..5564
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5465..5484
                     /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 ccggaccgcc atggcgaccc tggaaaagct gatgaaggcc ttcgagtccc tcaagtcctt
      661 ccagcagcag cagcagcagc agcagcagca gcagcagcag cagcagcagc agcagcagca
      721 gcagcagcag cagcagcagc agcagcagca gcagcagcag cagcagcagc agcagcagca
      781 gcagcagcag cagcagcagc agcagcagca gcagcagcaa cagccgccac cgccgccgcc
      841 gccgccgccg cctcctcagc ttcctcagcc gccgccgcag gcacagccgc tgctgcctca
      901 gccgcagccg cccccgccgc cgcccccgcc gccacccggc ccggctgtgg ctgaggagcc
      961 gctgcaccga ccaggctccc ttgtgagcaa gggcgaggag ctgttcaccg gggtggtgcc
     1021 catcctggtc gagctggacg gcgacgtaaa cggccacaag ttcagcgtgt ccggcgaggg
     1081 cgagggcgat gccacctacg gcaagctgac cctgaagctg atctgcacca ccggcaagct
     1141 gcccgtgccc tggcccaccc tcgtgaccac cctgggctac ggcctgcagt gcttcgcccg
     1201 ctaccccgac cacatgaagc agcacgactt cttcaagtcc gccatgcccg aaggctacgt
     1261 ccaggagcgc accatcttct tcaaggacga cggcaactac aagacccgcg ccgaggtgaa
     1321 gttcgagggc gacaccctgg tgaaccgcat cgagctgaag ggcatcgact tcaaggagga
     1381 cggcaacatc ctggggcaca agctggagta caactacaac agccacaacg tctatatcac
     1441 cgccgacaag cagaagaacg gcatcaaggc caacttcaag atccgccaca acatcgagga
     1501 cggcggcgtg cagctcgccg accactacca gcagaacacc cccatcggcg acggccccgt
     1561 gctgctgccc gacaaccact acctgagcta ccagtccaaa ctgagcaaag accccaacga
     1621 gaagcgcgat cacatggtcc tgctggagtt cgtgaccgcc gcctccggag ataacatggc
     1681 catcatcaag gagttcatgc gcttcaaggt gcacatggag ggctccgtga acggccacga
     1741 gttcgagatc gagggcgagg gcgagggccg cccctacgag ggcacccaga ccgccaagct
     1801 gaaggtgacc aagggtggcc ccctgccctt cgcctgggac atcctgtccc ctcagttcat
     1861 gtacggctcc aaggcctacg tgaagcaccc cgccgacatc cccgactact tgaagctgtc
     1921 cttccccgag ggcttcaagt gggagcgcgt gatgaacttc gaggacggcg gcgtggtgac
     1981 cgtgacccag gactcctccc tgcaggacgg cgagttcatc tacaaggtga agctgcgcgg
     2041 caccaacttc ccctccgacg gccccgtaat gcagaagaag accatgggct gggaggcctc
     2101 ctccgagcgg atgtaccccg aggacggcgc cctgaagggc gagatcaagc agaggctgaa
     2161 gctgaaggac ggcggccact acgacgctga ggtcaagacc acctacaagg ccaagaagcc
     2221 cgtgcagctg cccggcgcct acaacgtcaa catcaagttg gacatcacct cccacaacga
     2281 ggactacacc atcgtggaac agtacgaacg cgccgagggc cgccactcca ccggcggcat
     2341 gtaagcggcc gcgactctag atcataatca gccataccac atttgtagag gttttacttg
     2401 ctttaaaaaa cctcccacac ctccccctga acctgaaaca taaaatgaat gcaattgttg
     2461 ttgttaactt gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt
     2521 tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg
     2581 tatcttaagg cgtaaattgt aagcgttaat attttgttaa aattcgcgtt aaatttttgt
     2641 taaatcagct cattttttaa ccaataggcc gaaatcggca aaatccctta taaatcaaaa
     2701 gaatagaccg agatagggtt gagtgttgtt ccagtttgga acaagagtcc actattaaag
     2761 aacgtggact ccaacgtcaa agggcgaaaa accgtctatc agggcgatgg cccactacgt
     2821 gaaccatcac cctaatcaag ttttttgggg tcgaggtgcc gtaaagcact aaatcggaac
     2881 cctaaaggga gcccccgatt tagagcttga cggggaaagc cggcgaacgt ggcgagaaag
     2941 gaagggaaga aagcgaaagg agcgggcgct agggcgctgg caagtgtagc ggtcacgctg
     3001 cgcgtaacca ccacacccgc cgcgcttaat gcgccgctac agggcgcgtc aggtggcact
     3061 tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg
     3121 tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt
     3181 cctgaggcgg aaagaaccag ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag
     3241 gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accaggtgtg
     3301 gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag
     3361 caaccatagt cccgccccta actccgccca tcccgcccct aactccgccc agttccgccc
     3421 attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag gccgcctcgg
     3481 cctctgagct attccagaag tagtgaggag gcttttttgg aggcctaggc ttttgcaaag
     3541 atcgatcaag agacaggatg aggatcgttt cgcatgattg aacaagatgg attgcacgca
     3601 ggttctccgg ccgcttgggt ggagaggcta ttcggctatg actgggcaca acagacaatc
     3661 ggctgctctg atgccgccgt gttccggctg tcagcgcagg ggcgcccggt tctttttgtc
     3721 aagaccgacc tgtccggtgc cctgaatgaa ctgcaagacg aggcagcgcg gctatcgtgg
     3781 ctggccacga cgggcgttcc ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg
     3841 gactggctgc tattgggcga agtgccgggg caggatctcc tgtcatctca ccttgctcct
     3901 gccgagaaag tatccatcat ggctgatgca atgcggcggc tgcatacgct tgatccggct
     3961 acctgcccat tcgaccacca agcgaaacat cgcatcgagc gagcacgtac tcggatggaa
     4021 gccggtcttg tcgatcagga tgatctggac gaagagcatc aggggctcgc gccagccgaa
     4081 ctgttcgcca ggctcaaggc gagcatgccc gacggcgagg atctcgtcgt gacccatggc
     4141 gatgcctgct tgccgaatat catggtggaa aatggccgct tttctggatt catcgactgt
     4201 ggccggctgg gtgtggcgga ccgctatcag gacatagcgt tggctacccg tgatattgct
     4261 gaagagcttg gcggcgaatg ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc
     4321 gattcgcagc gcatcgcctt ctatcgcctt cttgacgagt tcttctgagc gggactctgg
     4381 ggttcgaaat gaccgaccaa gcgacgccca acctgccatc acgagatttc gattccaccg
     4441 ccgccttcta tgaaaggttg ggcttcggaa tcgttttccg ggacgccggc tggatgatcc
     4501 tccagcgcgg ggatctcatg ctggagttct tcgcccaccc tagggggagg ctaactgaaa
     4561 cacggaagga gacaataccg gaaggaaccc gcgctatgac ggcaataaaa agacagaata
     4621 aaacgcacgg tgttgggtcg tttgttcata aacgcggggt tcggtcccag ggctggcact
     4681 ctgtcgatac cccaccgaga ccccattggg gccaatacgc ccgcgtttct tccttttccc
     4741 caccccaccc cccaagttcg ggtgaaggcc cagggctcgc agccaacgtc ggggcggcag
     4801 gccctgccat agcctcaggt tactcatata tactttagat tgatttaaaa cttcattttt
     4861 aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac
     4921 gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag
     4981 atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg
     5041 tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca
     5101 gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac cacttcaaga
     5161 actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca
     5221 gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc
     5281 agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca
     5341 ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa
     5401 aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc
     5461 cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc
     5521 gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg
     5581 cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt cctgcgttat
     5641 cccctgattc tgtggataac cgtattaccg ccatgcat
//
