LOCUS       sequence-218415-        6147 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of human UBXD4 with C-terminal strep-HA tag.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6147)
  AUTHORS   Hulsmann J, Kravic B, Weith M, Gstaiger M, Aebersold RH, Collins BC,
            Meyer H
  TITLE     AP-SWATH reveals direct involvement of VCP/p97 in integrated stress 
            response signaling through facilitating CReP/PPP1R15B degradation.
  JOURNAL   Mol Cell Proteomics. 2018 Mar 29. pii: RA117.000471. doi: 
            10.1074/mcp.RA117.000471.
  PUBMED    29599191
REFERENCE   2  (bases 1 to 6147)
  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: "Mol Cell 
            Proteomics. 2018 Mar 29. pii: RA117.000471. doi: 
            10.1074/mcp.RA117.000471."
COMMENT     Sequence Label:  pcDNA5FRT/TO-UBXD4-Strep-HA
FEATURES             Location/Qualifiers
     source          1..6147
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        177..556
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        557..760
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     711..731
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     protein_bind    762..780
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    783..801
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     811..835
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     protein_bind    978..1002
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(1786..1810)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             1857..1880
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     CDS             1917..1940
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     CDS             1971..1997
                     /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             1971..1997
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     primer_bind     complement(2041..2058)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2047..2271
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(2404..2423)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     protein_bind    2555..2602
                     /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             2610..3632
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="KKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGY
                     VLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPE
                     TELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQT
                     VMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFG
                     DSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGN
                     FDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE
                     "
     CDS             2610..3632
                     /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    3762..3883
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3799..3818)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3853..3872
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3932..3948)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3932..3948)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3945..3967)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3956..3972
                     /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(3980..3986,3987..4004,4005..4010))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4025..4046
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4163..4180)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4334..4922)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4414..4433)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5093..5884,5885..5953))
                     /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     5716..5735
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5954..6058)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(join(6133..6147,1..5))
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 tgccgcatag ttaagccagt atctgctccc tgcttgtgtg ttggaggtcg ctgagtagtg
       61 cgcgagcaaa atttaagcta caacaaggca aggcttgacc gacaattgca tgaagaatct
      121 gcttagggtt aggcgttttg cgctgcttcg cgatgtacgg gccagatata cgcgttgaca
      181 ttgattattg actagttatt aatagtaatc aattacgggg tcattagttc atagcccata
      241 tatggagttc cgcgttacat aacttacggt aaatggcccg cctggctgac cgcccaacga
      301 cccccgccca ttgacgtcaa taatgacgta tgttcccata gtaacgccaa tagggacttt
      361 ccattgacgt caatgggtgg agtatttacg gtaaactgcc cacttggcag tacatcaagt
      421 gtatcatatg ccaagtacgc cccctattga cgtcaatgac ggtaaatggc ccgcctggca
      481 ttatgcccag tacatgacct tatgggactt tcctacttgg cagtacatct acgtattagt
      541 catcgctatt accatggtga tgcggttttg gcagtacatc aatgggcgtg gatagcggtt
      601 tgactcacgg ggatttccaa gtctccaccc cattgacgtc aatgggagtt tgttttggca
      661 ccaaaatcaa cgggactttc caaaatgtcg taacaactcc gccccattga cgcaaatggg
      721 cggtaggcgt gtacggtggg aggtctatat aagcagagct ctccctatca gtgatagaga
      781 tctccctatc agtgatagag atcgtcgacg agctcgttta gtgaaccgtc agatcgcctg
      841 gagacgccat ccacgctgtt ttgacctcca tagaagacac cgggaccgat ccagcctccg
      901 gactctagcg tttaaactta agcttggtac cgagctcgga tccactagtc cagtgtggtg
      961 gaattctgca gatatcaaca agtttgtaca aaaaagcagg ctccaccatg aaagacgtag
     1021 ataacctcaa aagtataaaa gaagaatggg tttgtgaaac aggatctgat aatcaacctc
     1081 ttggtaataa tcaacaatca aattgtgaat attttgttga tagccttttt gaggaagctc
     1141 agaaggttag ttccaaatgt gtgtctcccg ctgaacagaa gaaacaggta gatgtaaata
     1201 taaaattatg gaaaaacgga ttcaccgtca acgacgattt cagaagttat tccgatggtg
     1261 ccagtcagca gtttttgaac tccatcaaaa agggggaatt accttcagaa ttacagggaa
     1321 tttttgataa agaagaggtg gacgttaaag ttgaagacaa gaaaaatgaa atatgtttgt
     1381 ctacgaagcc tgtgttccag cccttttcag gacagggtca cagactagga agtgccacac
     1441 caaaaattgt ttctaaagca aagaatattg aagttgaaaa taaaaataat ttgtctgctg
     1501 ttccactgaa caacttggaa cccattacta atatacagat ctggttggcc aatggaaaaa
     1561 ggattgtcca gaaatttaac attactcata gagtaagcca tatcaaagac ttcattgaaa
     1621 aataccaagg atctcaaaga agtcctccgt tttccctggc aacagctctt cctgtcctca
     1681 ggttgctaga tgagacactc acactggaag aagcagattt acagaatgct gtcatcattc
     1741 agagactcca aaaaactgca tcttttagag aactttcaga gcacgaccca gctttcttgt
     1801 acaaagtggt tgatatcaag gccgccgaca tcaccagcct gtacaagaaa gtggggtgga
     1861 gccatcccca gttcgagaag ggcggaggat ctggcggcgg aagcggcgga ggaagctggt
     1921 cccaccctca gtttgagaaa ggcaccgagc tgggcagcac catggccagc tacccctacg
     1981 acgtgcccga ctacgcctga tgactcgagt ctagagggcc cgtttaaacc cgctgatcag
     2041 cctcgactgt gccttctagt tgccagccat ctgttgtttg cccctccccc gtgccttcct
     2101 tgaccctgga aggtgccact cccactgtcc tttcctaata aaatgaggaa attgcatcgc
     2161 attgtctgag taggtgtcat tctattctgg ggggtggggt ggggcaggac agcaaggggg
     2221 aggattggga agacaatagc aggcatgctg gggatgcggt gggctctatg gcttctgagg
     2281 cggaaagaac cagctggggc tctagggggt atccccacgc gccctgtagc ggcgcattaa
     2341 gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgccagc gccctagcgc
     2401 ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag
     2461 ctctaaatcg ggggctccct ttagggttcc gatttagtgc tttacggcac ctcgacccca
     2521 aaaaacttga ttagggtgat ggttcacgta cctagaagtt cctattccga agttcctatt
     2581 ctctagaaag tataggaact tccttggcca aaaagcctga actcaccgcg acgtctgtcg
     2641 agaagtttct gatcgaaaag ttcgacagcg tctccgacct gatgcagctc tcggagggcg
     2701 aagaatctcg tgctttcagc ttcgatgtag gagggcgtgg atatgtcctg cgggtaaata
     2761 gctgcgccga tggtttctac aaagatcgtt atgtttatcg gcactttgca tcggccgcgc
     2821 tcccgattcc ggaagtgctt gacattgggg aattcagcga gagcctgacc tattgcatct
     2881 cccgccgtgc acagggtgtc acgttgcaag acctgcctga aaccgaactg cccgctgttc
     2941 tgcagccggt cgcggaggcc atggatgcga tcgctgcggc cgatcttagc cagacgagcg
     3001 ggttcggccc attcggaccg caaggaatcg gtcaatacac tacatggcgt gatttcatat
     3061 gcgcgattgc tgatccccat gtgtatcact ggcaaactgt gatggacgac accgtcagtg
     3121 cgtccgtcgc gcaggctctc gatgagctga tgctttgggc cgaggactgc cccgaagtcc
     3181 ggcacctcgt gcacgcggat ttcggctcca acaatgtcct gacggacaat ggccgcataa
     3241 cagcggtcat tgactggagc gaggcgatgt tcggggattc ccaatacgag gtcgccaaca
     3301 tcttcttctg gaggccgtgg ttggcttgta tggagcagca gacgcgctac ttcgagcgga
     3361 ggcatccgga gcttgcagga tcgccgcggc tccgggcgta tatgctccgc attggtcttg
     3421 accaactcta tcagagcttg gttgacggca atttcgatga tgcagcttgg gcgcagggtc
     3481 gatgcgacgc aatcgtccga tccggagccg ggactgtcgg gcgtacacaa atcgcccgca
     3541 gaagcgcggc cgtctggacc gatggctgtg tagaagtact cgccgatagt ggaaaccgac
     3601 gccccagcac tcgtccgagg gcaaaggaat agcacgtact acgagatttc gattccaccg
     3661 ccgccttcta tgaaaggttg ggcttcggaa tcgttttccg ggacgccggc tggatgatcc
     3721 tccagcgcgg ggatctcatg ctggagttct tcgcccaccc caacttgttt attgcagctt
     3781 ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac
     3841 tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc tgtataccgt
     3901 cgacctctag ctagagcttg gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt
     3961 atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg
     4021 cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg
     4081 gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc
     4141 gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc
     4201 ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata
     4261 acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg
     4321 cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct
     4381 caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa
     4441 gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc
     4501 tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt
     4561 aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg
     4621 ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg
     4681 cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct
     4741 tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc
     4801 tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg
     4861 ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc
     4921 aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt
     4981 aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa
     5041 aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat
     5101 gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct
     5161 gactccccgt cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg
     5221 caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag
     5281 ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta
     5341 attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg
     5401 ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg
     5461 gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct
     5521 ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta
     5581 tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg
     5641 gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc
     5701 cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg
     5761 gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga
     5821 tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg
     5881 ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat
     5941 gttgaatact catactcttc ctttttcaat attattgaag catttatcag ggttattgtc
     6001 tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca
     6061 catttccccg aaaagtgcca cctgacgtcg acggatcggg agatctcccg atcccctatg
     6121 gtgcactctc agtacaatct gctctga
//
