LOCUS       sequence-353794-        7236 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  expresses mVenus and mCherry tagged DNAJB6b in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7236)
  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 7236)
  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: pVC-DNAJB6b
FEATURES             Location/Qualifiers
     source          1..7236
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(44..63)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        235..614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        615..818
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     769..789
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     protein_bind    820..838
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    841..859
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     869..893
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             996..1037
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             join(1050..1052,1053..1565)
                     /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(1092..1113)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1353..1372)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1700..1721
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1861..1879)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     2129..2148
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     complement(3188..3205)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3194..3418
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(3551..3570)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     protein_bind    3702..3749
                     /label=FRT
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="FLP-mediated recombination occurs in the 8-bp core 
                     sequence TCTAGAAA (Turan and Bode, 2011)."
     CDS             3757..4779
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="KKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGY
                     VLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPE
                     TELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQT
                     VMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFG
                     DSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGN
                     FDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE
                     "
     CDS             3757..4779
                     /codon_start=1
                     /gene="aph(4)-Ia"
                     /product="aminoglycoside phosphotransferase from E. coli"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="KKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGY
                     VLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPE
                     TELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQT
                     VMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFG
                     DSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGN
                     FDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE
                     "
     polyA_signal    4909..5030
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4946..4965)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5000..5019
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5079..5095)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5079..5095)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5092..5114)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5103..5119
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(5127..5133,5134..5151,5152..5157))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5172..5193
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5310..5327)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5481..6069)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5561..5580)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6240..7031,7032..7100))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     6863..6882
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7101..7205)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ccctatcagt gatagagatc
      841 tccctatcag tgatagagat cgtcgacgag ctcgtttagt gaaccgtcag atcgcctgga
      901 gacgccatcc acgctgtttt gacctccata gaagacaccg ggaccgatcc agcctccgga
      961 ctctagcgtt taaacttaag cttgccacct aaatgggtaa gcctatccct aaccctctcc
     1021 tcggtctcga ttctacggga tccggaaccg tgagcaaggg cgaggagctg ttcaccgggg
     1081 tggtgcccat cctggtcgag ctggacggcg acgtaaacgg ccacaagttc agcgtgtccg
     1141 gcgagggcga gggcgatgcc acctacggca agctgaccct gaagctgatc tgcaccaccg
     1201 gcaagctgcc cgtgccctgg cccaccctcg tgaccaccct gggctacggc ctgcagtgct
     1261 tcgcccgcta ccccgaccac atgaagcagc acgacttctt caagtccgcc atgcccgaag
     1321 gctacgtcca ggagcgcacc atcttcttca aggacgacgg caactacaag acccgcgccg
     1381 aggtgaagtt cgagggcgac accctggtga accgcatcga gctgaagggc atcgacttca
     1441 aggaggacgg caacatcctg gggcacaagc tggagtacaa ctacaacagc cacaacgtct
     1501 atatcaccgc cgacaagcag aagaacggca tcaaggccaa cttcaagatc cgccacaaca
     1561 tcgaggacgg cggcgtgcag ctcgccgacc actaccagca gaacaccccc atcggcgacg
     1621 gccccgtgct gctgcccgac aaccactacc tgagctacca gtccaaactg agcaaagacc
     1681 ccaacgagaa gcgcgatcac atggtcctgc tggagttcgt gaccgccgcc tccggagata
     1741 acatggccat catcaaggag ttcatgcgct tcaaggtgca catggagggc tccgtgaacg
     1801 gccacgagtt cgagatcgag ggcgagggcg agggccgccc ctacgagggc acccagaccg
     1861 ccaagctgaa ggtgaccaag ggtggccccc tgcccttcgc ctgggacatc ctgtcccctc
     1921 agttcatgta cggctccaag gcctacgtga agcaccccgc cgacatcccc gactacttga
     1981 agctgtcctt ccccgagggc ttcaagtggg agcgcgtgat gaacttcgag gacggcggcg
     2041 tggtgaccgt gacccaggac tcctccctgc aggacggcga gttcatctac aaggtgaagc
     2101 tgcgcggcac caacttcccc tccgacggcc ccgtaatgca gaagaagacc atgggctggg
     2161 aggcctcctc cgagcggatg taccccgagg acggcgccct gaagggcgag atcaagcaga
     2221 ggctgaagct gaaggacggc ggccactacg acgctgaggt caagaccacc tacaaggcca
     2281 agaagcccgt gcagctgccc ggcgcctaca acgtcaacat caagttggac atcacctccc
     2341 acaacgagga ctacaccatc gtggaacagt acgaacgcgc cgagggccgc cactccaccg
     2401 gcggcatggg ctcccttatg gtggattact atgaagttct aggcgtgcag agacatgcct
     2461 cacccgagga tattaaaaag gcatatcgga aactggcact gaagtggcat ccagataaaa
     2521 atcctgagaa taaagaagaa gcagagagaa aattcaagca agtagcggag gcatatgaag
     2581 tgctgtcgga tgctaagaaa cgggacatct atgacaaata tggcaaagaa ggattaaatg
     2641 gtggaggagg aggtggaagt cattttgaca gtccatttga atttggcttc acattccgta
     2701 acccagatga tgtcttcagg gaattttttg gtggaaggga cccattttca tttgacttct
     2761 ttgaagaccc ttttgaggac ttctttggga atcgaagggg tccccgagga agcagaagcc
     2821 gagggacggg gtcgtttttc tctgcgttca gtggatttcc gtcttttgga agtggatttt
     2881 cttcttttga tacaggattt acttcatttg ggtcactagg tcacgggggc ctcacttcat
     2941 tctcttccac gtcatttggt ggtagtggca tgggcaactt caaatcgata tcaacttcaa
     3001 ctaaaatggt taatggcaga aaaatcacta caaagagaat tgtcgagaac ggtcaagaaa
     3061 gagtagaagt tgaagaagat ggccagttaa agtccttaac aataaatggt aaggagcagc
     3121 tgctgcgctt ggataacaag taagcggccg ctcgagtcta gagggcccgt ttaaacccgc
     3181 tgatcagcct cgactgtgcc ttctagttgc cagccatctg ttgtttgccc ctcccccgtg
     3241 ccttccttga ccctggaagg tgccactccc actgtccttt cctaataaaa tgaggaaatt
     3301 gcatcgcatt gtctgagtag gtgtcattct attctggggg gtggggtggg gcaggacagc
     3361 aagggggagg attgggaaga caatagcagg catgctgggg atgcggtggg ctctatggct
     3421 tctgaggcgg aaagaaccag ctggggctct agggggtatc cccacgcgcc ctgtagcggc
     3481 gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact tgccagcgcc
     3541 ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc cggctttccc
     3601 cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagtgcttt acggcacctc
     3661 gaccccaaaa aacttgatta gggtgatggt tcacgtacct agaagttcct attccgaagt
     3721 tcctattctc tagaaagtat aggaacttcc ttggccaaaa agcctgaact caccgcgacg
     3781 tctgtcgaga agtttctgat cgaaaagttc gacagcgtct ccgacctgat gcagctctcg
     3841 gagggcgaag aatctcgtgc tttcagcttc gatgtaggag ggcgtggata tgtcctgcgg
     3901 gtaaatagct gcgccgatgg tttctacaaa gatcgttatg tttatcggca ctttgcatcg
     3961 gccgcgctcc cgattccgga agtgcttgac attggggaat tcagcgagag cctgacctat
     4021 tgcatctccc gccgtgcaca gggtgtcacg ttgcaagacc tgcctgaaac cgaactgccc
     4081 gctgttctgc agccggtcgc ggaggccatg gatgcgatcg ctgcggccga tcttagccag
     4141 acgagcgggt tcggcccatt cggaccgcaa ggaatcggtc aatacactac atggcgtgat
     4201 ttcatatgcg cgattgctga tccccatgtg tatcactggc aaactgtgat ggacgacacc
     4261 gtcagtgcgt ccgtcgcgca ggctctcgat gagctgatgc tttgggccga ggactgcccc
     4321 gaagtccggc acctcgtgca cgcggatttc ggctccaaca atgtcctgac ggacaatggc
     4381 cgcataacag cggtcattga ctggagcgag gcgatgttcg gggattccca atacgaggtc
     4441 gccaacatct tcttctggag gccgtggttg gcttgtatgg agcagcagac gcgctacttc
     4501 gagcggaggc atccggagct tgcaggatcg ccgcggctcc gggcgtatat gctccgcatt
     4561 ggtcttgacc aactctatca gagcttggtt gacggcaatt tcgatgatgc agcttgggcg
     4621 cagggtcgat gcgacgcaat cgtccgatcc ggagccggga ctgtcgggcg tacacaaatc
     4681 gcccgcagaa gcgcggccgt ctggaccgat ggctgtgtag aagtactcgc cgatagtgga
     4741 aaccgacgcc ccagcactcg tccgagggca aaggaatagc acgtactacg agatttcgat
     4801 tccaccgccg ccttctatga aaggttgggc ttcggaatcg ttttccggga cgccggctgg
     4861 atgatcctcc agcgcgggga tctcatgctg gagttcttcg cccaccccaa cttgtttatt
     4921 gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt
     4981 ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta tcatgtctgt
     5041 ataccgtcga cctctagcta gagcttggcg taatcatggt catagctgtt tcctgtgtga
     5101 aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa gtgtaaagcc
     5161 tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact gcccgctttc
     5221 cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc
     5281 ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt
     5341 cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca
     5401 ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa
     5461 aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat
     5521 cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc
     5581 cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc
     5641 gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt
     5701 tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac
     5761 cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg
     5821 ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca
     5881 gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc
     5941 gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa
     6001 accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa
     6061 ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac
     6121 tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta
     6181 aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt
     6241 taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata
     6301 gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc
     6361 agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac
     6421 cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag
     6481 tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac
     6541 gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc
     6601 agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg
     6661 gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc
     6721 atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct
     6781 gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc
     6841 tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc
     6901 atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc
     6961 agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc
     7021 gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca
     7081 cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt
     7141 tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt
     7201 ccgcgcacat ttccccgaaa agtgccacct gacgtc
//
