LOCUS       sequence-372207-        7602 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  MAC-tagged gene expression of human DUX4.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7602)
  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 7602)
  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
FEATURES             Location/Qualifiers
     source          1..7602
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(25..42)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(196..784)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(276..295)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(955..1746,1747..1815))
                     /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     1578..1597
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1816..1920)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(1995..2014)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        2186..2565
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2566..2769
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2720..2740
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     protein_bind    2771..2789
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    2792..2810
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     2820..2844
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     protein_bind    2987..3011
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(4290..4314)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             4361..4384
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     CDS             4421..4444
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     CDS             4475..4501
                     /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             4475..4501
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             join(4502..4849,4850..4852,4853..5461)
                     /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(5505..5522)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    5511..5735
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(5868..5887)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     protein_bind    6019..6066
                     /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             6074..7096
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="KKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGY
                     VLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPE
                     TELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQT
                     VMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFG
                     DSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGN
                     FDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE
                     "
     CDS             6074..7096
                     /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    7226..7347
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(7263..7282)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     7317..7336
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(7396..7412)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7396..7412)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(7409..7431)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    7420..7436
                     /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(7444..7450,7451..7468,7469..7474))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7489..7510
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
ORIGIN
        1 gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct
       61 gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga
      121 taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc
      181 cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg
      241 ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg
      301 aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt
      361 tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt
      421 gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg
      481 cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact
      541 ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt
      601 cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct
      661 gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac
      721 cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc
      781 tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg
      841 ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta
      901 aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca
      961 atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc
     1021 ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc
     1081 tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc
     1141 agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat
     1201 taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt
     1261 tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc
     1321 cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag
     1381 ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt
     1441 tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac
     1501 tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg
     1561 cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat
     1621 tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc
     1681 gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc
     1741 tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa
     1801 atgttgaata ctcatactct tcctttttca atattattga agcatttatc agggttattg
     1861 tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg
     1921 cacatttccc cgaaaagtgc cacctgacgt cgacggatcg ggagatctcc cgatccccta
     1981 tggtgcactc tcagtacaat ctgctctgat gccgcatagt taagccagta tctgctccct
     2041 gcttgtgtgt tggaggtcgc tgagtagtgc gcgagcaaaa tttaagctac aacaaggcaa
     2101 ggcttgaccg acaattgcat gaagaatctg cttagggtta ggcgttttgc gctgcttcgc
     2161 gatgtacggg ccagatatac gcgttgacat tgattattga ctagttatta atagtaatca
     2221 attacggggt cattagttca tagcccatat atggagttcc gcgttacata acttacggta
     2281 aatggcccgc ctggctgacc gcccaacgac ccccgcccat tgacgtcaat aatgacgtat
     2341 gttcccatag taacgccaat agggactttc cattgacgtc aatgggtgga gtatttacgg
     2401 taaactgccc acttggcagt acatcaagtg tatcatatgc caagtacgcc ccctattgac
     2461 gtcaatgacg gtaaatggcc cgcctggcat tatgcccagt acatgacctt atgggacttt
     2521 cctacttggc agtacatcta cgtattagtc atcgctatta ccatggtgat gcggttttgg
     2581 cagtacatca atgggcgtgg atagcggttt gactcacggg gatttccaag tctccacccc
     2641 attgacgtca atgggagttt gttttggcac caaaatcaac gggactttcc aaaatgtcgt
     2701 aacaactccg ccccattgac gcaaatgggc ggtaggcgtg tacggtggga ggtctatata
     2761 agcagagctc tccctatcag tgatagagat ctccctatca gtgatagaga tcgtcgacga
     2821 gctcgtttag tgaaccgtca gatcgcctgg agacgccatc cacgctgttt tgacctccat
     2881 agaagacacc gggaccgatc cagcctccgg actctagcgt ttaaacttaa gcttggtacc
     2941 gagctcggat ccactagtcc agtgtggtgg aattctgcag atatcaacaa gtttgtacaa
     3001 aaaagcaggc tccaccatgg ccctcccgac accctcggac agcaccctcc ccgcggaagc
     3061 ccggggacga ggacggcgac ggagactcgt ttggaccccg agccaaagcg aggccctgcg
     3121 agcctgcttt gagcggaacc cgtacccggg catcgccacc agagaacggc tggcccaggc
     3181 catcggcatt ccggagccca gggtccagat ttggtttcag aatgagaggt cacgccagct
     3241 gaggcagcac cggcgggaat ctcggccctg gcccgggaga cgcggcccgc cagaaggccg
     3301 gcgaaagcgg accgccgtca ccggatccca gaccgccctg ctcctccgag cctttgagaa
     3361 ggatcgcttt ccaggcatcg ccgcccggga ggagctggcc agagagacgg gcctcccgga
     3421 gtccaggatt cagatctggt ttcagaatcg aagggccagg cacccgggac agggtggcag
     3481 ggcgcccgcg caggcaggcg gcctgtgcag cgcggccccc ggcgggggtc accctgctcc
     3541 ctcgtgggtc gccttcgccc acaccggcgc gtggggaacg gggcttcccg caccccacgt
     3601 gccctgcgcg cctggggctc tcccacaggg ggctttcgtg agccaggcag cgagggccgc
     3661 ccccgcgctg cagcccagcc aggccgcgcc ggcagagggg atctcccaac ctgccccggc
     3721 gcgcggggat ttcgcctacg ccgccccggc tcctccggac ggggcgctct cccaccctca
     3781 ggctcctcgc tggcctccgc acccgggcaa aagccgggag gaccgggacc cgcagcgcga
     3841 cggcctgccg ggcccctgcg cggtggcaca gcctgggccc gctcaagcgg ggccgcaggg
     3901 ccaaggggtg cttgcgccac ccacgtccca ggggagtccg tggtggggct ggggccgggg
     3961 tccccaggtc gccggggcgg cgtgggaacc ccaagccggg gcagctccac ctccccagcc
     4021 cgcgcccccg gacgcctccg cctccgcgcg gcaggggcag atgcaaggca tcccggcgcc
     4081 ctcccaggcg ctccaggagc cggcgccctg gtctgcactc ccctgcggcc tgctgctgga
     4141 tgagctcctg gcgagcccgg agtttctgca gcaggcgcaa cctctcctag aaacggaggc
     4201 cccgggggag ctggaggcct cggaagaggc cgcctcgctg gaagcacccc tcagcgagga
     4261 agaataccgg gctctgctgg aggagcttga cccagctttc ttgtacaaag tggttgatat
     4321 caaggccgcc gacatcacca gcctgtacaa gaaagtgggg tggagccatc cccagttcga
     4381 gaagggcgga ggatctggcg gcggaagcgg cggaggaagc tggtcccacc ctcagtttga
     4441 gaaaggcacc gagctgggca gcaccatggc cagctacccc tacgacgtgc ccgactacgc
     4501 caaggacaac accgtgcccc tgaagctgat cgccctgctg gccaacggcg agttccactc
     4561 tggcgagcag ctgggcgaga cactgggcat gagcagagcc gccatcaaca agcacatcca
     4621 gaccctgcgg gactggggcg tggacgtgtt cacagtgccc ggcaagggct acagcctgcc
     4681 cgagcccatc cagctgctga acgccaagca gatcctgggc cagctggatg gcggatctgt
     4741 ggccgtgctg cccgtgatcg acagcaccaa ccagtatctg ctcgaccgga tcggcgagct
     4801 gaagtccggc gacgcctgta tcgccgagta ccagcaggcc ggcagaggcg gcaggggcag
     4861 aaagtggttc agccccttcg gcgccaacct gtacctgagc atgttctgga gactggaaca
     4921 gggcccagcc gccgctattg gcctgagcct ggtcatcggc atcgtgatgg ccgaggtgct
     4981 gcggaagctg ggcgccgaca aagtgcgcgt gaagtggccc aacgacctgt atctgcagga
     5041 ccggaagctg gccggcatcc tggtggaact gaccggcaaa acaggcgacg ccgcccagat
     5101 cgtgatcggc gctggcatca acatggccat gcggcgggtg gaagagagcg tggtcaacca
     5161 gggctggatc accctgcagg aagccggcat caacctggac cggaacaccc tggccgccat
     5221 gctgatcaga gagctgagag ccgccctgga actgttcgag caggaaggcc tggcccccta
     5281 cctgagcaga tgggagaagc tggacaactt catcaaccgg cccgtgaaac tgatcatcgg
     5341 cgacaaagag atcttcggca tcagccgggg catcgacaag cagggggccc tgctcctgga
     5401 acaggacggc atcatcaagc cctggatggg cggcgagatc agcctgcgga gcgccgagaa
     5461 gtgatgactc gagtctagag ggcccgttta aacccgctga tcagcctcga ctgtgccttc
     5521 tagttgccag ccatctgttg tttgcccctc ccccgtgcct tccttgaccc tggaaggtgc
     5581 cactcccact gtcctttcct aataaaatga ggaaattgca tcgcattgtc tgagtaggtg
     5641 tcattctatt ctggggggtg gggtggggca ggacagcaag ggggaggatt gggaagacaa
     5701 tagcaggcat gctggggatg cggtgggctc tatggcttct gaggcggaaa gaaccagctg
     5761 gggctctagg gggtatcccc acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt
     5821 ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt
     5881 cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcgggggct
     5941 ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgattaggg
     6001 tgatggttca cgtacctaga agttcctatt ccgaagttcc tattctctag aaagtatagg
     6061 aacttccttg gccaaaaagc ctgaactcac cgcgacgtct gtcgagaagt ttctgatcga
     6121 aaagttcgac agcgtctccg acctgatgca gctctcggag ggcgaagaat ctcgtgcttt
     6181 cagcttcgat gtaggagggc gtggatatgt cctgcgggta aatagctgcg ccgatggttt
     6241 ctacaaagat cgttatgttt atcggcactt tgcatcggcc gcgctcccga ttccggaagt
     6301 gcttgacatt ggggaattca gcgagagcct gacctattgc atctcccgcc gtgcacaggg
     6361 tgtcacgttg caagacctgc ctgaaaccga actgcccgct gttctgcagc cggtcgcgga
     6421 ggccatggat gcgatcgctg cggccgatct tagccagacg agcgggttcg gcccattcgg
     6481 accgcaagga atcggtcaat acactacatg gcgtgatttc atatgcgcga ttgctgatcc
     6541 ccatgtgtat cactggcaaa ctgtgatgga cgacaccgtc agtgcgtccg tcgcgcaggc
     6601 tctcgatgag ctgatgcttt gggccgagga ctgccccgaa gtccggcacc tcgtgcacgc
     6661 ggatttcggc tccaacaatg tcctgacgga caatggccgc ataacagcgg tcattgactg
     6721 gagcgaggcg atgttcgggg attcccaata cgaggtcgcc aacatcttct tctggaggcc
     6781 gtggttggct tgtatggagc agcagacgcg ctacttcgag cggaggcatc cggagcttgc
     6841 aggatcgccg cggctccggg cgtatatgct ccgcattggt cttgaccaac tctatcagag
     6901 cttggttgac ggcaatttcg atgatgcagc ttgggcgcag ggtcgatgcg acgcaatcgt
     6961 ccgatccgga gccgggactg tcgggcgtac acaaatcgcc cgcagaagcg cggccgtctg
     7021 gaccgatggc tgtgtagaag tactcgccga tagtggaaac cgacgcccca gcactcgtcc
     7081 gagggcaaag gaatagcacg tactacgaga tttcgattcc accgccgcct tctatgaaag
     7141 gttgggcttc ggaatcgttt tccgggacgc cggctggatg atcctccagc gcggggatct
     7201 catgctggag ttcttcgccc accccaactt gtttattgca gcttataatg gttacaaata
     7261 aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg
     7321 tttgtccaaa ctcatcaatg tatcttatca tgtctgtata ccgtcgacct ctagctagag
     7381 cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc
     7441 acacaacata cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta
     7501 actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca
     7561 gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt tt
//
