LOCUS       sequence-218406-        6705 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of human UBXD8 with C-terminal strep-HA tag.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6705)
  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 6705)
  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-UBXD8-Strep-HA
FEATURES             Location/Qualifiers
     source          1..6705
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        99..478
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        479..682
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     633..653
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     protein_bind    684..702
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    705..723
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     733..757
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     protein_bind    900..924
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     regulatory      928..937
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    complement(2266..2290)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             2337..2360
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     CDS             2397..2420
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     CDS             2451..2477
                     /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             2451..2477
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     primer_bind     complement(2521..2538)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2527..2751
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(2884..2903)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     protein_bind    3035..3082
                     /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             3090..4112
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="KKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGY
                     VLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPE
                     TELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQT
                     VMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFG
                     DSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGN
                     FDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE
                     "
     CDS             3090..4112
                     /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    4242..4363
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4279..4298)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4333..4352
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4412..4428)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4412..4428)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4425..4447)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4436..4452
                     /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(4460..4466,4467..4484,4485..4490))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4505..4526
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4643..4660)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4814..5402)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4894..4913)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5573..6364,6365..6433))
                     /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     6196..6215
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6434..6538)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(6613..6632)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 tacaacaagg caaggcttga ccgacaattg catgaagaat ctgcttaggg ttaggcgttt
       61 tgcgctgctt cgcgatgtac gggccagata tacgcgttga cattgattat tgactagtta
      121 ttaatagtaa tcaattacgg ggtcattagt tcatagccca tatatggagt tccgcgttac
      181 ataacttacg gtaaatggcc cgcctggctg accgcccaac gacccccgcc cattgacgtc
      241 aataatgacg tatgttccca tagtaacgcc aatagggact ttccattgac gtcaatgggt
      301 ggagtattta cggtaaactg cccacttggc agtacatcaa gtgtatcata tgccaagtac
      361 gccccctatt gacgtcaatg acggtaaatg gcccgcctgg cattatgccc agtacatgac
      421 cttatgggac tttcctactt ggcagtacat ctacgtatta gtcatcgcta ttaccatggt
      481 gatgcggttt tggcagtaca tcaatgggcg tggatagcgg tttgactcac ggggatttcc
      541 aagtctccac cccattgacg tcaatgggag tttgttttgg caccaaaatc aacgggactt
      601 tccaaaatgt cgtaacaact ccgccccatt gacgcaaatg ggcggtaggc gtgtacggtg
      661 ggaggtctat ataagcagag ctctccctat cagtgataga gatctcccta tcagtgatag
      721 agatcgtcga cgagctcgtt tagtgaaccg tcagatcgcc tggagacgcc atccacgctg
      781 ttttgacctc catagaagac accgggaccg atccagcctc cggactctag cgtttaaact
      841 taagcttggt accgagctcg gatccactag tccagtgtgg tggaattctg cagatatcaa
      901 caagtttgta caaaaaagca ggctccacca tggcggcgcc tgaggagcgg gatctaaccc
      961 aggagcagac agagaagctg ctgcagtttc aggatctcac tggcatcgaa tctatggatc
     1021 agtgtcgcca taccttggaa cagcataact ggaacataga ggctgctgta caggacagat
     1081 tgaatgagca agagggcgta cctagtgttt tcaacccacc tccatcacga cccctgcagg
     1141 ttaatacagc tgaccacagg atctacagct atgttgtctc aagacctcaa ccaagggggc
     1201 tgcttggatg gggttattac ttgataatgc ttccattccg gtttacctat tacacgatac
     1261 ttgatatatt taggtttgct cttcgtttta tacggcctga ccctcgcagc cgggtcactg
     1321 accccgttgg ggacattgtt tcatttatgc actcttttga agagaaatat gggagggcac
     1381 accctgtctt ctaccaggga acgtacagcc aggcacttaa cgatgccaaa agggagcttc
     1441 gctttctttt ggtttatctt catggagatg atcaccagga ctctgatgag ttttgtcgca
     1501 acacactctg tgcacctgaa gttatttcac taataaacac taggatgctc ttctgggcat
     1561 gctctacaaa caaacctgag ggatacaggg tctcacaggc tttacgagag aacacctatc
     1621 cattcctggc catgattatg ctgaaggatc gaaggatgac tgtggtggga cggctagaag
     1681 gcctcattca acctgatgac ctcattaacc aactgacatt tatcatggat gctaaccaga
     1741 cttacctggt gtcagaacgc ctagaaaggg aagaaagaaa ccagacccaa gtgctgagac
     1801 aacagcagga tgaggcctac ctggcctctc tcagagctga ccaggagaaa gaaagaaaga
     1861 aacgggagga gcgggagcgt aagcggcgga aggaggagga ggtgcaacag caaaagttgg
     1921 cagaggagag acggcggcag aatttacagg aggaaaagga aaggaagttg gaatgcctgc
     1981 cccctgaacc ttcccctgat gaccctgaaa gtgtcaagat catcttcaaa ttacctaatg
     2041 attctcgagt agagagacga ttccactttt cacagtctct aacagtaatc cacgacttct
     2101 tattctcctt gaaggaaagc ccagaaaagt ttcagattga agccaatttt cccaggcgag
     2161 tgctgccctg catcccttca gaggagtggc ccaatccccc tacgctacag gaggccggac
     2221 tcagccacac agaagttctt tttgttcagg acctaactga cgaataccca gctttcttgt
     2281 acaaagtggt tgatatcaag gccgccgaca tcaccagcct gtacaagaaa gtggggtgga
     2341 gccatcccca gttcgagaag ggcggaggat ctggcggcgg aagcggcgga ggaagctggt
     2401 cccaccctca gtttgagaaa ggcaccgagc tgggcagcac catggccagc tacccctacg
     2461 acgtgcccga ctacgcctga tgactcgagt ctagagggcc cgtttaaacc cgctgatcag
     2521 cctcgactgt gccttctagt tgccagccat ctgttgtttg cccctccccc gtgccttcct
     2581 tgaccctgga aggtgccact cccactgtcc tttcctaata aaatgaggaa attgcatcgc
     2641 attgtctgag taggtgtcat tctattctgg ggggtggggt ggggcaggac agcaaggggg
     2701 aggattggga agacaatagc aggcatgctg gggatgcggt gggctctatg gcttctgagg
     2761 cggaaagaac cagctggggc tctagggggt atccccacgc gccctgtagc ggcgcattaa
     2821 gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgccagc gccctagcgc
     2881 ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag
     2941 ctctaaatcg ggggctccct ttagggttcc gatttagtgc tttacggcac ctcgacccca
     3001 aaaaacttga ttagggtgat ggttcacgta cctagaagtt cctattccga agttcctatt
     3061 ctctagaaag tataggaact tccttggcca aaaagcctga actcaccgcg acgtctgtcg
     3121 agaagtttct gatcgaaaag ttcgacagcg tctccgacct gatgcagctc tcggagggcg
     3181 aagaatctcg tgctttcagc ttcgatgtag gagggcgtgg atatgtcctg cgggtaaata
     3241 gctgcgccga tggtttctac aaagatcgtt atgtttatcg gcactttgca tcggccgcgc
     3301 tcccgattcc ggaagtgctt gacattgggg aattcagcga gagcctgacc tattgcatct
     3361 cccgccgtgc acagggtgtc acgttgcaag acctgcctga aaccgaactg cccgctgttc
     3421 tgcagccggt cgcggaggcc atggatgcga tcgctgcggc cgatcttagc cagacgagcg
     3481 ggttcggccc attcggaccg caaggaatcg gtcaatacac tacatggcgt gatttcatat
     3541 gcgcgattgc tgatccccat gtgtatcact ggcaaactgt gatggacgac accgtcagtg
     3601 cgtccgtcgc gcaggctctc gatgagctga tgctttgggc cgaggactgc cccgaagtcc
     3661 ggcacctcgt gcacgcggat ttcggctcca acaatgtcct gacggacaat ggccgcataa
     3721 cagcggtcat tgactggagc gaggcgatgt tcggggattc ccaatacgag gtcgccaaca
     3781 tcttcttctg gaggccgtgg ttggcttgta tggagcagca gacgcgctac ttcgagcgga
     3841 ggcatccgga gcttgcagga tcgccgcggc tccgggcgta tatgctccgc attggtcttg
     3901 accaactcta tcagagcttg gttgacggca atttcgatga tgcagcttgg gcgcagggtc
     3961 gatgcgacgc aatcgtccga tccggagccg ggactgtcgg gcgtacacaa atcgcccgca
     4021 gaagcgcggc cgtctggacc gatggctgtg tagaagtact cgccgatagt ggaaaccgac
     4081 gccccagcac tcgtccgagg gcaaaggaat agcacgtact acgagatttc gattccaccg
     4141 ccgccttcta tgaaaggttg ggcttcggaa tcgttttccg ggacgccggc tggatgatcc
     4201 tccagcgcgg ggatctcatg ctggagttct tcgcccaccc caacttgttt attgcagctt
     4261 ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac
     4321 tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc tgtataccgt
     4381 cgacctctag ctagagcttg gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt
     4441 atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg
     4501 cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg
     4561 gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc
     4621 gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc
     4681 ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata
     4741 acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg
     4801 cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct
     4861 caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa
     4921 gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc
     4981 tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt
     5041 aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg
     5101 ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg
     5161 cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct
     5221 tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc
     5281 tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg
     5341 ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc
     5401 aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt
     5461 aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa
     5521 aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat
     5581 gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct
     5641 gactccccgt cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg
     5701 caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag
     5761 ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta
     5821 attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg
     5881 ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg
     5941 gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct
     6001 ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta
     6061 tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg
     6121 gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc
     6181 cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg
     6241 gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga
     6301 tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg
     6361 ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat
     6421 gttgaatact catactcttc ctttttcaat attattgaag catttatcag ggttattgtc
     6481 tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca
     6541 catttccccg aaaagtgcca cctgacgtcg acggatcggg agatctcccg atcccctatg
     6601 gtgcactctc agtacaatct gctctgatgc cgcatagtta agccagtatc tgctccctgc
     6661 ttgtgtgttg gaggtcgctg agtagtgcgc gagcaaaatt taagc
//
