LOCUS       sequence-362344-        5591 bp    DNA     circular SYN 14-APR-2025
DEFINITION  expresses mVenus and mCherry tagged to a mutant of the huntingtin 
            exon1 (polyQ25) in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5591)
  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 5591)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 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: pQ25-VC
FEATURES             Location/Qualifiers
     source          1..5591
                     /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(896..898,899..1411)
                     /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(938..959)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1199..1218)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1546..1567
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1707..1725)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     1975..1994
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     polyA_signal    2379..2500
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2416..2435)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2470..2489
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2507..2962)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2644..2665)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2856..2875
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2989..3093
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3095..3452
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3095..3286
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3118..3138)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3303..3438
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3487..4281
                     /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             3487..4281
                     /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(3541..3560)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4151..4170
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4469..4488)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4513..4560
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4889..5477
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5378..5397
                     /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 caacagccgc caccgccgcc gccgccgccg ccgcctcctc agcttcctca
      781 gccgccgccg caggcacagc cgctgctgcc tcagccgcag ccgcccccgc cgccgccccc
      841 gccgccaccc ggcccggctg tggctgagga gccgctgcac cgaccaggct cccttgtgag
      901 caagggcgag gagctgttca ccggggtggt gcccatcctg gtcgagctgg acggcgacgt
      961 aaacggccac aagttcagcg tgtccggcga gggcgagggc gatgccacct acggcaagct
     1021 gaccctgaag ctgatctgca ccaccggcaa gctgcccgtg ccctggccca ccctcgtgac
     1081 caccctgggc tacggcctgc agtgcttcgc ccgctacccc gaccacatga agcagcacga
     1141 cttcttcaag tccgccatgc ccgaaggcta cgtccaggag cgcaccatct tcttcaagga
     1201 cgacggcaac tacaagaccc gcgccgaggt gaagttcgag ggcgacaccc tggtgaaccg
     1261 catcgagctg aagggcatcg acttcaagga ggacggcaac atcctggggc acaagctgga
     1321 gtacaactac aacagccaca acgtctatat caccgccgac aagcagaaga acggcatcaa
     1381 ggccaacttc aagatccgcc acaacatcga ggacggcggc gtgcagctcg ccgaccacta
     1441 ccagcagaac acccccatcg gcgacggccc cgtgctgctg cccgacaacc actacctgag
     1501 ctaccagtcc aaactgagca aagaccccaa cgagaagcgc gatcacatgg tcctgctgga
     1561 gttcgtgacc gccgcctccg gagataacat ggccatcatc aaggagttca tgcgcttcaa
     1621 ggtgcacatg gagggctccg tgaacggcca cgagttcgag atcgagggcg agggcgaggg
     1681 ccgcccctac gagggcaccc agaccgccaa gctgaaggtg accaagggtg gccccctgcc
     1741 cttcgcctgg gacatcctgt cccctcagtt catgtacggc tccaaggcct acgtgaagca
     1801 ccccgccgac atccccgact acttgaagct gtccttcccc gagggcttca agtgggagcg
     1861 cgtgatgaac ttcgaggacg gcggcgtggt gaccgtgacc caggactcct ccctgcagga
     1921 cggcgagttc atctacaagg tgaagctgcg cggcaccaac ttcccctccg acggccccgt
     1981 aatgcagaag aagaccatgg gctgggaggc ctcctccgag cggatgtacc ccgaggacgg
     2041 cgccctgaag ggcgagatca agcagaggct gaagctgaag gacggcggcc actacgacgc
     2101 tgaggtcaag accacctaca aggccaagaa gcccgtgcag ctgcccggcg cctacaacgt
     2161 caacatcaag ttggacatca cctcccacaa cgaggactac accatcgtgg aacagtacga
     2221 acgcgccgag ggccgccact ccaccggcgg catgtaagcg gccgcgactc tagatcataa
     2281 tcagccatac cacatttgta gaggttttac ttgctttaaa aaacctccca cacctccccc
     2341 tgaacctgaa acataaaatg aatgcaattg ttgttgttaa cttgtttatt gcagcttata
     2401 atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc
     2461 attctagttg tggtttgtcc aaactcatca atgtatctta aggcgtaaat tgtaagcgtt
     2521 aatattttgt taaaattcgc gttaaatttt tgttaaatca gctcattttt taaccaatag
     2581 gccgaaatcg gcaaaatccc ttataaatca aaagaataga ccgagatagg gttgagtgtt
     2641 gttccagttt ggaacaagag tccactatta aagaacgtgg actccaacgt caaagggcga
     2701 aaaaccgtct atcagggcga tggcccacta cgtgaaccat caccctaatc aagttttttg
     2761 gggtcgaggt gccgtaaagc actaaatcgg aaccctaaag ggagcccccg atttagagct
     2821 tgacggggaa agccggcgaa cgtggcgaga aaggaaggga agaaagcgaa aggagcgggc
     2881 gctagggcgc tggcaagtgt agcggtcacg ctgcgcgtaa ccaccacacc cgccgcgctt
     2941 aatgcgccgc tacagggcgc gtcaggtggc acttttcggg gaaatgtgcg cggaacccct
     3001 atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga
     3061 taaatgcttc aataatattg aaaaaggaag agtcctgagg cggaaagaac cagctgtgga
     3121 atgtgtgtca gttagggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa
     3181 gcatgcatct caattagtca gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca
     3241 gaagtatgca aagcatgcat ctcaattagt cagcaaccat agtcccgccc ctaactccgc
     3301 ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt
     3361 tttttattta tgcagaggcc gaggccgcct cggcctctga gctattccag aagtagtgag
     3421 gaggcttttt tggaggccta ggcttttgca aagatcgatc aagagacagg atgaggatcg
     3481 tttcgcatga ttgaacaaga tggattgcac gcaggttctc cggccgcttg ggtggagagg
     3541 ctattcggct atgactgggc acaacagaca atcggctgct ctgatgccgc cgtgttccgg
     3601 ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg acctgtccgg tgccctgaat
     3661 gaactgcaag acgaggcagc gcggctatcg tggctggcca cgacgggcgt tccttgcgca
     3721 gctgtgctcg acgttgtcac tgaagcggga agggactggc tgctattggg cgaagtgccg
     3781 gggcaggatc tcctgtcatc tcaccttgct cctgccgaga aagtatccat catggctgat
     3841 gcaatgcggc ggctgcatac gcttgatccg gctacctgcc cattcgacca ccaagcgaaa
     3901 catcgcatcg agcgagcacg tactcggatg gaagccggtc ttgtcgatca ggatgatctg
     3961 gacgaagagc atcaggggct cgcgccagcc gaactgttcg ccaggctcaa ggcgagcatg
     4021 cccgacggcg aggatctcgt cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg
     4081 gaaaatggcc gcttttctgg attcatcgac tgtggccggc tgggtgtggc ggaccgctat
     4141 caggacatag cgttggctac ccgtgatatt gctgaagagc ttggcggcga atgggctgac
     4201 cgcttcctcg tgctttacgg tatcgccgct cccgattcgc agcgcatcgc cttctatcgc
     4261 cttcttgacg agttcttctg agcgggactc tggggttcga aatgaccgac caagcgacgc
     4321 ccaacctgcc atcacgagat ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg
     4381 gaatcgtttt ccgggacgcc ggctggatga tcctccagcg cggggatctc atgctggagt
     4441 tcttcgccca ccctaggggg aggctaactg aaacacggaa ggagacaata ccggaaggaa
     4501 cccgcgctat gacggcaata aaaagacaga ataaaacgca cggtgttggg tcgtttgttc
     4561 ataaacgcgg ggttcggtcc cagggctggc actctgtcga taccccaccg agaccccatt
     4621 ggggccaata cgcccgcgtt tcttcctttt ccccacccca ccccccaagt tcgggtgaag
     4681 gcccagggct cgcagccaac gtcggggcgg caggccctgc catagcctca ggttactcat
     4741 atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc
     4801 tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag
     4861 accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct
     4921 gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac
     4981 caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc
     5041 tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg
     5101 ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt
     5161 tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt
     5221 gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc
     5281 tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca
     5341 gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata
     5401 gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg
     5461 ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct
     5521 ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta
     5581 ccgccatgca t
//
