LOCUS       sequence-214208-        6472 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of human p47 with C-terminal strep-HA tag.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6472)
  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 6472)
  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-p47-Strep-HA
FEATURES             Location/Qualifiers
     source          1..6472
                     /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"
     protein_bind    1036..1060
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(2171..2195)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             2242..2265
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     CDS             2302..2325
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     CDS             2356..2382
                     /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             2356..2382
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     primer_bind     complement(2425..2442)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2431..2655
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(2788..2807)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     protein_bind    2939..2986
                     /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             2994..4016
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="KKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGY
                     VLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPE
                     TELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQT
                     VMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFG
                     DSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGN
                     FDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE
                     "
     CDS             2994..4016
                     /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    4146..4267
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4183..4202)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4237..4256
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4316..4332)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4316..4332)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4329..4351)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4340..4356
                     /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(4364..4370,4371..4388,4389..4394))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4409..4430
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4547..4564)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4718..5306)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4798..4817)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(5775..6152)
                     /label=ISS
                     /note="immunostimulatory sequence from the AmpR gene; 
                     contains unmethylated CpG dinucleotides in the context of 
                     5'-AACGTT-3' (Sato et al., 1996)"
     primer_bind     6099..6118
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6337..6441)
                     /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 cttggtaccg agctcggatc cactagtcca gtgtggtgga
     1021 attctgcaga tatcaacaag tttgtacaaa aaagttggca tggcggcgga gcgacaggag
     1081 gcgctgaggg agttcgtggc ggtgacgggc gccgaggagg accgggcccg cttctttctc
     1141 gagtcggccg gctgggactt gcagatcgcg ctagcgagct tttatgagga cggaggggat
     1201 gaagacattg tgaccatttc gcaggcaacc cccagttcag tgtccagagg cacagccccc
     1261 agtgataata gagtgacatc cttcagagac ctcattcatg accaagatga agatgaggag
     1321 gaagaggaag gccagaggtt ttatgctggg ggctcagaga gaagtggaca gcagattgtt
     1381 ggccctccca ggaagaaaag tcccaacgag ctggtggatg atctctttaa aggtgccaaa
     1441 gagcatggag ctgtagctgt ggagcgagtg accaagagcc ctggagagac cagtaaaccg
     1501 agaccatttg caggaggtgg ctaccgcctt ggggcagcac cagaggaaga gtctgcctat
     1561 gtggcaggag aaaagaggca gcattccagc caagatgttc atgtagtatt gaaactctgg
     1621 aagagtggat tcagcctgga taatggagaa ctcagaagct accaagaccc atccaatgcc
     1681 cagtttctgg agtctatccg cagaggggag gtgccagcag agcttcggag gctagctcac
     1741 ggtggacagg tgaacttgga tatggaggac catcgggacg aggactttgt gaagcccaaa
     1801 ggagccttca aagccttcac tggcgagggt cagaaactgg gcagcactgc cccccaggtg
     1861 ttgagtacca gctctccagc ccaacaggca gaaaatgaag ccaaagccag ctcttccatc
     1921 ttaatcgacg aatcagagcc taccacaaac atccaaattc ggcttgcaga cggcgggagg
     1981 ctggtgcaga aatttaacca cagccacagg atcagcgaca tccgactctt catcgtggat
     2041 gcccggccag ccatggctgc caccagcttt atcctcatga ctactttccc gaacaaagag
     2101 ctggctgatg agagccagac cctgaaggaa gccaacctgc tcaatgctgt catcgtgcag
     2161 cggttaacat acccaacttt cttgtacaaa gtggttgata tcaaggccgc cgacatcacc
     2221 agcctgtaca agaaagtggg gtggagccat ccccagttcg agaagggcgg aggatctggc
     2281 ggcggaagcg gcggaggaag ctggtcccac cctcagtttg agaaaggcac cgagctgggc
     2341 agcaccatgg ccagctaccc ctacgacgtg cccgactacg cctgatgact cgagtctaga
     2401 gggcccgtta aacccgctga tcagcctcga ctgtgccttc tagttgccag ccatctgttg
     2461 tttgcccctc ccccgtgcct tccttgaccc tggaaggtgc cactcccact gtcctttcct
     2521 aataaaatga ggaaattgca tcgcattgtc tgagtaggtg tcattctatt ctggggggtg
     2581 gggtggggca ggacagcaag ggggaggatt gggaagacaa tagcaggcat gctggggatg
     2641 cggtgggctc tatggcttct gaggcggaaa gaaccagctg gggctctagg gggtatcccc
     2701 acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg
     2761 ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca
     2821 cgttcgccgg ctttccccgt caagctctaa atcgggggct ccctttaggg ttccgattta
     2881 gtgctttacg gcacctcgac cccaaaaaac ttgattaggg tgatggttca cgtacctaga
     2941 agttcctatt ccgaagttcc tattctctag aaagtatagg aacttccttg gccaaaaagc
     3001 ctgaactcac cgcgacgtct gtcgagaagt ttctgatcga aaagttcgac agcgtctccg
     3061 acctgatgca gctctcggag ggcgaagaat ctcgtgcttt cagcttcgat gtaggagggc
     3121 gtggatatgt cctgcgggta aatagctgcg ccgatggttt ctacaaagat cgttatgttt
     3181 atcggcactt tgcatcggcc gcgctcccga ttccggaagt gcttgacatt ggggaattca
     3241 gcgagagcct gacctattgc atctcccgcc gtgcacaggg tgtcacgttg caagacctgc
     3301 ctgaaaccga actgcccgct gttctgcagc cggtcgcgga ggccatggat gcgatcgctg
     3361 cggccgatct tagccagacg agcgggttcg gcccattcgg accgcaagga atcggtcaat
     3421 acactacatg gcgtgatttc atatgcgcga ttgctgatcc ccatgtgtat cactggcaaa
     3481 ctgtgatgga cgacaccgtc agtgcgtccg tcgcgcaggc tctcgatgag ctgatgcttt
     3541 gggccgagga ctgccccgaa gtccggcacc tcgtgcacgc ggatttcggc tccaacaatg
     3601 tcctgacgga caatggccgc ataacagcgg tcattgactg gagcgaggcg atgttcgggg
     3661 attcccaata cgaggtcgcc aacatcttct tctggaggcc gtggttggct tgtatggagc
     3721 agcagacgcg ctacttcgag cggaggcatc cggagcttgc aggatcgccg cggctccggg
     3781 cgtatatgct ccgcattggt cttgaccaac tctatcagag cttggttgac ggcaatttcg
     3841 atgatgcagc ttgggcgcag ggtcgatgcg acgcaatcgt ccgatccgga gccgggactg
     3901 tcgggcgtac acaaatcgcc cgcagaagcg cggccgtctg gaccgatggc tgtgtagaag
     3961 tactcgccga tagtggaaac cgacgcccca gcactcgtcc gagggcaaag gaatagcacg
     4021 tactacgaga tttcgattcc accgccgcct tctatgaaag gttgggcttc ggaatcgttt
     4081 tccgggacgc cggctggatg atcctccagc gcggggatct catgctggag ttcttcgccc
     4141 accccaactt gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt
     4201 tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg
     4261 tatcttatca tgtctgtata ccgtcgacct ctagctagag cttggcgtaa tcatggtcat
     4321 agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa
     4381 gcataaagtg taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc
     4441 gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc
     4501 aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact
     4561 cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac
     4621 ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa
     4681 aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg
     4741 acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa
     4801 gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc
     4861 ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac
     4921 gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac
     4981 cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg
     5041 taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt
     5101 atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa
     5161 cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct
     5221 cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga
     5281 ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg
     5341 ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct
     5401 tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt
     5461 aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc
     5521 tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg
     5581 gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag
     5641 atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt
     5701 tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag
     5761 ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt
     5821 ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca
     5881 tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg
     5941 ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat
     6001 ccgtagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat
     6061 gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag
     6121 aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt
     6181 accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc
     6241 ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa
     6301 gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg
     6361 aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa
     6421 taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tc
//
