LOCUS       sequence-372499-        7584 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  DUX4 with KBM (p. E414-L424 deletion) with MAC-C tag.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7584)
  AUTHORS   Salokas K, Liu X, Ohman T, Chowdhury I, Gawriyski L, Keskitalo S, 
            Varjosalo M
  TITLE     Physical and functional interactome atlas of human receptor tyrosine
            kinases.
  JOURNAL   EMBO Rep. 2022 Apr 5:e54041. doi: 10.15252/embr.202154041.
  PUBMED    35384245
REFERENCE   2  (bases 1 to 7584)
  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: "EMBO Rep. 
            2022 Apr 5:e54041. doi: 10.15252/embr.202154041."
COMMENT     Sequence Label: MAC_C_DUX4_KBM
FEATURES             Location/Qualifiers
     source          1..7584
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(join(528..1319,1320..1388))
                     /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     1151..1170
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1389..1493)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(1568..1587)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        1759..2138
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2139..2342
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2293..2313
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     protein_bind    2344..2362
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2365..2383
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     2393..2417
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     protein_bind    2560..2584
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(3845..3869)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             3916..3939
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     CDS             3976..3999
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     CDS             4030..4056
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /note="This is a variant of the standard HA tag, which has 
                     sequence YPYDVPDYA. Some anti-HA antibodies may not 
                     recognize this variant."
                     /translation="YPYDVPDYA"
     CDS             4030..4056
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             join(4057..4404,4405..4407,4408..5016)
                     /codon_start=1
                     /product="promiscuous R118G mutant of the E. coli biotin 
                     protein ligase (Cronan, 2005)"
                     /label=BirA(R118G)
                     /note="also known as BirA* or BioID"
                     /translation="KDNTVPLKLIALLANGEFHSGEQLGETLGMSRAAINKHIQTLRDW
                     GVDVFTVPGKGYSLPEPIQLLNAKQILGQLDGGSVAVLPVIDSTNQYLLDRIGELKSGD
                     ACIAEYQQAGRGGRGRKWFSPFGANLYLSMFWRLEQGPAAAIGLSLVIGIVMAEVLRKL
                     GADKVRVKWPNDLYLQDRKLAGILVELTGKTGDAAQIVIGAGINMAMRRVEESVVNQGW
                     ITLQEAGINLDRNTLAAMLIRELRAALELFEQEGLAPYLSRWEKLDNFINRPVKLIIGD
                     KEIFGISRGIDKQGALLLEQDGIIKPWMGGEISLRSAEK"
     primer_bind     complement(5060..5077)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    5066..5290
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(5423..5442)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     protein_bind    5574..5621
                     /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             5629..6651
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="KKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGY
                     VLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPE
                     TELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQT
                     VMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFG
                     DSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGN
                     FDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE
                     "
     CDS             5629..6651
                     /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    6781..6902
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6818..6837)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6872..6891
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(6951..6967)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6951..6967)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6964..6986)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6975..6991
                     /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(6999..7005,7006..7023,7024..7029))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7044..7065
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(7182..7199)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(join(7353..7584,1..357))
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7433..7452)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta
       61 tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca
      121 gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag
      181 tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag
      241 ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt
      301 agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa
      361 gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg
      421 attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga
      481 agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta
      541 atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc
      601 cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg
      661 ataccgcgag acccacgctc accggctcca gatttatcag caataaacca gccagccgga
      721 agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt
      781 tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt
      841 gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc
      901 caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc
      961 ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca
     1021 gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag
     1081 tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg
     1141 tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa
     1201 cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa
     1261 cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga
     1321 gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga
     1381 atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg
     1441 agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt
     1501 ccccgaaaag tgccacctga cgtcgacgga tcgggagatc tcccgatccc ctatggtgca
     1561 ctctcagtac aatctgctct gatgccgcat agttaagcca gtatctgctc cctgcttgtg
     1621 tgttggaggt cgctgagtag tgcgcgagca aaatttaagc tacaacaagg caaggcttga
     1681 ccgacaattg catgaagaat ctgcttaggg ttaggcgttt tgcgctgctt cgcgatgtac
     1741 gggccagata tacgcgttga cattgattat tgactagtta ttaatagtaa tcaattacgg
     1801 ggtcattagt tcatagccca tatatggagt tccgcgttac ataacttacg gtaaatggcc
     1861 cgcctggctg accgcccaac gacccccgcc cattgacgtc aataatgacg tatgttccca
     1921 tagtaacgcc aatagggact ttccattgac gtcaatgggt ggagtattta cggtaaactg
     1981 cccacttggc agtacatcaa gtgtatcata tgccaagtac gccccctatt gacgtcaatg
     2041 acggtaaatg gcccgcctgg cattatgccc agtacatgac cttatgggac tttcctactt
     2101 ggcagtacat ctacgtatta gtcatcgcta ttaccatggt gatgcggttt tggcagtaca
     2161 tcaatgggcg tggatagcgg tttgactcac ggggatttcc aagtctccac cccattgacg
     2221 tcaatgggag tttgttttgg caccaaaatc aacgggactt tccaaaatgt cgtaacaact
     2281 ccgccccatt gacgcaaatg ggcggtaggc gtgtacggtg ggaggtctat ataagcagag
     2341 ctctccctat cagtgataga gatctcccta tcagtgatag agatcgtcga cgagctcgtt
     2401 tagtgaaccg tcagatcgcc tggagacgcc atccacgctg ttttgacctc catagaagac
     2461 accgggaccg atccagcctc cggactctag cgtttaaact taagcttggt accgagctcg
     2521 gatccactag tccagtgtgg tggaattctg cagatatcaa caagtttgta caaaaaagca
     2581 ggctccacca tggccctccc gacaccctcg gacagcaccc tccccgcgga agcccgggga
     2641 cgaggacggc gacggagact cgtttggacc ccgagccaaa gcgaggccct gcgagcctgc
     2701 tttgagcgga acccgtaccc gggcatcgcc accagagaac ggctggccca ggccatcggc
     2761 attccggagc ccagggtcca gatttggttt cagaatgaga ggtcacgcca gctgaggcag
     2821 caccggcggg aatctcggcc ctggcccggg agacgcggcc cgccagaagg ccggcgaaag
     2881 cggaccgccg tcaccggatc ccagaccgcc ctgctcctcc gagcctttga gaaggatcgc
     2941 tttccaggca tcgccgcccg ggaggagctg gccagagaga cgggcctccc ggagtccagg
     3001 attcagatct ggtttcagaa tcgaagggcc aggcacccgg gacagggtgg cagggcgccc
     3061 gcgcaggcag gcggcctgtg cagcgcggcc cccggcgggg gtcaccctgc tccctcgtgg
     3121 gtcgccttcg cccacaccgg cgcgtgggga acggggcttc ccgcacccca cgtgccctgc
     3181 gcgcctgggg ctctcccaca gggggctttc gtgagccagg cagcgagggc cgcccccgcg
     3241 ctgcagccca gccaggccgc gccggcagag gggatctccc aacctgcccc ggcgcgcggg
     3301 gatttcgcct acgccgcccc ggctcctccg gacggggcgc tctcccaccc tcaggctcct
     3361 cgctggcctc cgcacccggg caaaagccgg gaggaccggg acccgcagcg cgacggcctg
     3421 ccgggcccct gcgcggtggc acagcctggg cccgctcaag cggggccgca gggccaaggg
     3481 gtgcttgcgc cacccacgtc ccaggggagt ccgtggtggg gctggggccg gggtccccag
     3541 gtcgccgggg cggcgtggga accccaagcc ggggcagctc cacctcccca gcccgcgccc
     3601 ccggacgcct ccgcctccgc gcggcagggg cagatgcaag gcatcccggc gccctcccag
     3661 gcgctccagg agccggcgcc ctggtctgca ctcccctgcg gcctgctgct ggatgagctc
     3721 ctggcgagcc cggagtttct gcagcaggcg caacctctcc tagaaacgga ggccccgggg
     3781 gagctggagg cctcggaaga ggccgcctcg ctggaagcac ccctcagcga ggaagaggag
     3841 cttgacccag ctttcttgta caaagtggtt gatatcaagg ccgccgacat caccagcctg
     3901 tacaagaaag tggggtggag ccatccccag ttcgagaagg gcggaggatc tggcggcgga
     3961 agcggcggag gaagctggtc ccaccctcag tttgagaaag gcaccgagct gggcagcacc
     4021 atggccagct acccctacga cgtgcccgac tacgccaagg acaacaccgt gcccctgaag
     4081 ctgatcgccc tgctggccaa cggcgagttc cactctggcg agcagctggg cgagacactg
     4141 ggcatgagca gagccgccat caacaagcac atccagaccc tgcgggactg gggcgtggac
     4201 gtgttcacag tgcccggcaa gggctacagc ctgcccgagc ccatccagct gctgaacgcc
     4261 aagcagatcc tgggccagct ggatggcgga tctgtggccg tgctgcccgt gatcgacagc
     4321 accaaccagt atctgctcga ccggatcggc gagctgaagt ccggcgacgc ctgtatcgcc
     4381 gagtaccagc aggccggcag aggcggcagg ggcagaaagt ggttcagccc cttcggcgcc
     4441 aacctgtacc tgagcatgtt ctggagactg gaacagggcc cagccgccgc tattggcctg
     4501 agcctggtca tcggcatcgt gatggccgag gtgctgcgga agctgggcgc cgacaaagtg
     4561 cgcgtgaagt ggcccaacga cctgtatctg caggaccgga agctggccgg catcctggtg
     4621 gaactgaccg gcaaaacagg cgacgccgcc cagatcgtga tcggcgctgg catcaacatg
     4681 gccatgcggc gggtggaaga gagcgtggtc aaccagggct ggatcaccct gcaggaagcc
     4741 ggcatcaacc tggaccggaa caccctggcc gccatgctga tcagagagct gagagccgcc
     4801 ctggaactgt tcgagcagga aggcctggcc ccctacctga gcagatggga gaagctggac
     4861 aacttcatca accggcccgt gaaactgatc atcggcgaca aagagatctt cggcatcagc
     4921 cggggcatcg acaagcaggg ggccctgctc ctggaacagg acggcatcat caagccctgg
     4981 atgggcggcg agatcagcct gcggagcgcc gagaagtgat gactcgagtc tagagggccc
     5041 gtttaaaccc gctgatcagc ctcgactgtg ccttctagtt gccagccatc tgttgtttgc
     5101 ccctcccccg tgccttcctt gaccctggaa ggtgccactc ccactgtcct ttcctaataa
     5161 aatgaggaaa ttgcatcgca ttgtctgagt aggtgtcatt ctattctggg gggtggggtg
     5221 gggcaggaca gcaaggggga ggattgggaa gacaatagca ggcatgctgg ggatgcggtg
     5281 ggctctatgg cttctgaggc ggaaagaacc agctggggct ctagggggta tccccacgcg
     5341 ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca
     5401 cttgccagcg ccctagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc
     5461 gccggctttc cccgtcaagc tctaaatcgg gggctccctt tagggttccg atttagtgct
     5521 ttacggcacc tcgaccccaa aaaacttgat tagggtgatg gttcacgtac ctagaagttc
     5581 ctattccgaa gttcctattc tctagaaagt ataggaactt ccttggccaa aaagcctgaa
     5641 ctcaccgcga cgtctgtcga gaagtttctg atcgaaaagt tcgacagcgt ctccgacctg
     5701 atgcagctct cggagggcga agaatctcgt gctttcagct tcgatgtagg agggcgtgga
     5761 tatgtcctgc gggtaaatag ctgcgccgat ggtttctaca aagatcgtta tgtttatcgg
     5821 cactttgcat cggccgcgct cccgattccg gaagtgcttg acattgggga attcagcgag
     5881 agcctgacct attgcatctc ccgccgtgca cagggtgtca cgttgcaaga cctgcctgaa
     5941 accgaactgc ccgctgttct gcagccggtc gcggaggcca tggatgcgat cgctgcggcc
     6001 gatcttagcc agacgagcgg gttcggccca ttcggaccgc aaggaatcgg tcaatacact
     6061 acatggcgtg atttcatatg cgcgattgct gatccccatg tgtatcactg gcaaactgtg
     6121 atggacgaca ccgtcagtgc gtccgtcgcg caggctctcg atgagctgat gctttgggcc
     6181 gaggactgcc ccgaagtccg gcacctcgtg cacgcggatt tcggctccaa caatgtcctg
     6241 acggacaatg gccgcataac agcggtcatt gactggagcg aggcgatgtt cggggattcc
     6301 caatacgagg tcgccaacat cttcttctgg aggccgtggt tggcttgtat ggagcagcag
     6361 acgcgctact tcgagcggag gcatccggag cttgcaggat cgccgcggct ccgggcgtat
     6421 atgctccgca ttggtcttga ccaactctat cagagcttgg ttgacggcaa tttcgatgat
     6481 gcagcttggg cgcagggtcg atgcgacgca atcgtccgat ccggagccgg gactgtcggg
     6541 cgtacacaaa tcgcccgcag aagcgcggcc gtctggaccg atggctgtgt agaagtactc
     6601 gccgatagtg gaaaccgacg ccccagcact cgtccgaggg caaaggaata gcacgtacta
     6661 cgagatttcg attccaccgc cgccttctat gaaaggttgg gcttcggaat cgttttccgg
     6721 gacgccggct ggatgatcct ccagcgcggg gatctcatgc tggagttctt cgcccacccc
     6781 aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca
     6841 aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct
     6901 tatcatgtct gtataccgtc gacctctagc tagagcttgg cgtaatcatg gtcatagctg
     6961 tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc cggaagcata
     7021 aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca
     7081 ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc
     7141 gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg
     7201 cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta
     7261 tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc
     7321 aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag
     7381 catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac
     7441 caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc
     7501 ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt
     7561 aggtatctca gttcggtgta ggtc
//
