LOCUS       sequence-218570-        7899 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of human Cezanne with C-terminal strep-HA tag.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7899)
  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 7899)
  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-Cezanne-Strep-HA
FEATURES             Location/Qualifiers
     source          1..7899
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        149..528
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        529..732
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     683..703
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     protein_bind    734..752
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    755..773
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     783..807
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     protein_bind    950..974
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(3510..3534)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             3581..3604
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     CDS             3641..3664
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     CDS             3695..3721
                     /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             3695..3721
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     primer_bind     complement(3765..3782)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3771..3995
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(4128..4147)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     protein_bind    4279..4326
                     /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             4334..5356
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="KKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGY
                     VLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPE
                     TELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQT
                     VMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFG
                     DSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGN
                     FDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE
                     "
     CDS             4334..5356
                     /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    5486..5607
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5523..5542)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5577..5596
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5656..5672)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5656..5672)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5669..5691)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5680..5696
                     /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(5704..5710,5711..5728,5729..5734))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5749..5770
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5887..5904)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6058..6646)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6138..6157)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6817..7608,7609..7677))
                     /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     7440..7459
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7678..7782)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(7857..7876)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 cctgcttgtg tgttggaggt cgctgagtag tgcgcgagca aaatttaagc tacaacaagg
       61 caaggcttga ccgacaattg catgaagaat ctgcttaggg ttaggcgttt tgcgctgctt
      121 cgcgatgtac gggccagata tacgcgttga cattgattat tgactagtta ttaatagtaa
      181 tcaattacgg ggtcattagt tcatagccca tatatggagt tccgcgttac ataacttacg
      241 gtaaatggcc cgcctggctg accgcccaac gacccccgcc cattgacgtc aataatgacg
      301 tatgttccca tagtaacgcc aatagggact ttccattgac gtcaatgggt ggagtattta
      361 cggtaaactg cccacttggc agtacatcaa gtgtatcata tgccaagtac gccccctatt
      421 gacgtcaatg acggtaaatg gcccgcctgg cattatgccc agtacatgac cttatgggac
      481 tttcctactt ggcagtacat ctacgtatta gtcatcgcta ttaccatggt gatgcggttt
      541 tggcagtaca tcaatgggcg tggatagcgg tttgactcac ggggatttcc aagtctccac
      601 cccattgacg tcaatgggag tttgttttgg caccaaaatc aacgggactt tccaaaatgt
      661 cgtaacaact ccgccccatt gacgcaaatg ggcggtaggc gtgtacggtg ggaggtctat
      721 ataagcagag ctctccctat cagtgataga gatctcccta tcagtgatag agatcgtcga
      781 cgagctcgtt tagtgaaccg tcagatcgcc tggagacgcc atccacgctg ttttgacctc
      841 catagaagac accgggaccg atccagcctc cggactctag cgtttaaact taagcttggt
      901 accgagctcg gatccactag tccagtgtgg tggaattctg cagatatcaa caagtttgta
      961 caaaaaagca ggctccacca tgaccctgga catggatgct gttctgtcag attttgtccg
     1021 ttccacagga gcagagccag ggctagcgcg agatctccta gaaggaaaga attgggatgt
     1081 gaatgccgcc ctcagtgatt ttgaacagct acgtcaagtc catgctggaa acctaccccc
     1141 atcctttagt gaggggagtg gtggctccag gacccctgaa aaagggtttt ctgacagaga
     1201 gcctactcgc cctccccgac ccatcctcca gcggcaggat gacatcgttc aagaaaaacg
     1261 cctgtctagg ggcatctccc acgccagctc cagcattgtt tccctggccc ggtcccatgt
     1321 ctcctccaat ggtgggggtg gggggagcaa tgagcacccc ctggaaatgc ccatctgtgc
     1381 cttccagctt ccagatctca ctgtatacaa tgaagacttc cgcagcttca tagagagaga
     1441 cctcattgag cagtccatgc tggttgcctt ggaacaggca gggcgtttga actggtgggt
     1501 gagtgtggat cccacctctc agaggctgct tcctttggca actactggag atgggaactg
     1561 cctcctgcat gcagcctccc ttggaatgtg gggtttccat gatcgggact tgatgctgcg
     1621 gaaagctttg tatgcactga tggagaaggg agttgagaag gaagcgttga aaaggcgctg
     1681 gaggtggcag cagacacagc agaataaaga gtcagggctg gtatacacag aagatgaatg
     1741 gcagaaggag tggaatgaac tgatcaagct tgcctcaagt gaaccccgaa tgcatctagg
     1801 taccaatgga gccaactgtg gtggggtgga gagttctgag gagcctgtat atgagagcct
     1861 tgaagagttt cacgtctttg tccttgctca tgtgcttagg aggcccatag tcgtcgtggc
     1921 agacaccatg ctgagggact ccggagggga agcatttgcc cctattccct ttggaggaat
     1981 ctatctgcct ttggaggtcc cagccagcca gtgtcaccgc tcccctctgg tgctcgccta
     2041 tgatcaggcc cacttttctg cactcgtgtc catggagcag aaggagaata ccaaggaaca
     2101 agctgtgatc ccacttacag attcagagta taagctgctg cccttgcact ttgctgtgga
     2161 ccctggaaag ggctgggagt ggggcaaaga tgatagtgac aatgtccgat tggccagtgt
     2221 aattctgtcc ctagaggtca aattgcatct gctgcatagc tacatgaatg tgaagtggat
     2281 cccactgtcc tctgatgcac aggctcctct ggcccagcct gagtccccca ccgcctcagc
     2341 tggagatgag ccccggtcca ctcctgagtc tggagactca gacaaggagt cagttggcag
     2401 cagttccacc agcaacgagg gcggccggcg gaaggagaag tcaaagcgag atcgggagaa
     2461 ggacaagaag agagcagatt ctgtggctaa caaactgggc agctttggca aaaccttggg
     2521 cagcaagctc aagaagaaca tggggggcct gatgcacagc aagggttcaa agcctggagg
     2581 ggtggggaca gggttgggag gaagcagcgg cactgagaca ctggagaaga agaagaaaaa
     2641 ctcactgaag agctggaagg gtggcaagga ggaggcagct ggggatgggc ctgtgtctga
     2701 gaagccccca gctgagtctg ttggtaacgg agggagcaag tatagccagg aggtgatgca
     2761 gagcctgagc attctgagga ctgccatgca aggggagggg aagtttattt ttgttggaac
     2821 cctgaagatg ggtcaccgtc accagtatca ggaggaaatg atccagcgct acctttctga
     2881 tgctgaggag agattcctgg cagaacagaa gcagaaggag gcagagagga agatcatgaa
     2941 tggaggaata gggggtggcc ctcctccagc caaaaagcca gagccagatg ctagggaaga
     3001 gcagccgacc ggtcccccag cagagtccag ggcaatggca ttttccactg gctaccctgg
     3061 ggactttact atccctcggc cgtctggggg cggagtccac tgccaggaac cccggaggca
     3121 gttggcaggg ggtccatgtg tcgggggcct accaccatat gccaccttcc ccagacagtg
     3181 ccctcctggg cgaccctacc cccaccagga cagcatccct tctctggagc caggcagcca
     3241 ctctaaggat ggacttcaca ggggtgcctt gttaccaccc ccctaccgag tggctgattc
     3301 ctatagcaat ggctacagag agccccctga gccagatgga tgggctggag gtctccgggg
     3361 ccttccccca actcagacca aatgcaaaca accgaactgc agcttctatg gacaccctga
     3421 gacaaacaac ttctgttcct gttgttacag ggaagaactg aggaggaggg agcgggaacc
     3481 ggatggggag ctcctggtgc acaggttcga cccagctttc ttgtacaaag tggttgatat
     3541 caaggccgcc gacatcacca gcctgtacaa gaaagtgggg tggagccatc cccagttcga
     3601 gaagggcgga ggatctggcg gcggaagcgg cggaggaagc tggtcccacc ctcagtttga
     3661 gaaaggcacc gagctgggca gcaccatggc cagctacccc tacgacgtgc ccgactacgc
     3721 ctgatgactc gagtctagag ggcccgttta aacccgctga tcagcctcga ctgtgccttc
     3781 tagttgccag ccatctgttg tttgcccctc ccccgtgcct tccttgaccc tggaaggtgc
     3841 cactcccact gtcctttcct aataaaatga ggaaattgca tcgcattgtc tgagtaggtg
     3901 tcattctatt ctggggggtg gggtggggca ggacagcaag ggggaggatt gggaagacaa
     3961 tagcaggcat gctggggatg cggtgggctc tatggcttct gaggcggaaa gaaccagctg
     4021 gggctctagg gggtatcccc acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt
     4081 ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt
     4141 cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcgggggct
     4201 ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgattaggg
     4261 tgatggttca cgtacctaga agttcctatt ccgaagttcc tattctctag aaagtatagg
     4321 aacttccttg gccaaaaagc ctgaactcac cgcgacgtct gtcgagaagt ttctgatcga
     4381 aaagttcgac agcgtctccg acctgatgca gctctcggag ggcgaagaat ctcgtgcttt
     4441 cagcttcgat gtaggagggc gtggatatgt cctgcgggta aatagctgcg ccgatggttt
     4501 ctacaaagat cgttatgttt atcggcactt tgcatcggcc gcgctcccga ttccggaagt
     4561 gcttgacatt ggggaattca gcgagagcct gacctattgc atctcccgcc gtgcacaggg
     4621 tgtcacgttg caagacctgc ctgaaaccga actgcccgct gttctgcagc cggtcgcgga
     4681 ggccatggat gcgatcgctg cggccgatct tagccagacg agcgggttcg gcccattcgg
     4741 accgcaagga atcggtcaat acactacatg gcgtgatttc atatgcgcga ttgctgatcc
     4801 ccatgtgtat cactggcaaa ctgtgatgga cgacaccgtc agtgcgtccg tcgcgcaggc
     4861 tctcgatgag ctgatgcttt gggccgagga ctgccccgaa gtccggcacc tcgtgcacgc
     4921 ggatttcggc tccaacaatg tcctgacgga caatggccgc ataacagcgg tcattgactg
     4981 gagcgaggcg atgttcgggg attcccaata cgaggtcgcc aacatcttct tctggaggcc
     5041 gtggttggct tgtatggagc agcagacgcg ctacttcgag cggaggcatc cggagcttgc
     5101 aggatcgccg cggctccggg cgtatatgct ccgcattggt cttgaccaac tctatcagag
     5161 cttggttgac ggcaatttcg atgatgcagc ttgggcgcag ggtcgatgcg acgcaatcgt
     5221 ccgatccgga gccgggactg tcgggcgtac acaaatcgcc cgcagaagcg cggccgtctg
     5281 gaccgatggc tgtgtagaag tactcgccga tagtggaaac cgacgcccca gcactcgtcc
     5341 gagggcaaag gaatagcacg tactacgaga tttcgattcc accgccgcct tctatgaaag
     5401 gttgggcttc ggaatcgttt tccgggacgc cggctggatg atcctccagc gcggggatct
     5461 catgctggag ttcttcgccc accccaactt gtttattgca gcttataatg gttacaaata
     5521 aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg
     5581 tttgtccaaa ctcatcaatg tatcttatca tgtctgtata ccgtcgacct ctagctagag
     5641 cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc
     5701 acacaacata cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta
     5761 actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca
     5821 gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc
     5881 cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc
     5941 tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat
     6001 gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt
     6061 ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg
     6121 aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc
     6181 tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt
     6241 ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa
     6301 gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta
     6361 tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa
     6421 caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa
     6481 ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt
     6541 cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt
     6601 ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat
     6661 cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat
     6721 gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc
     6781 aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc
     6841 acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta
     6901 gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga
     6961 cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg
     7021 cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc
     7081 tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat
     7141 cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag
     7201 gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat
     7261 cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa
     7321 ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa
     7381 gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga
     7441 taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg
     7501 gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc
     7561 acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg
     7621 aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact
     7681 cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat
     7741 atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt
     7801 gccacctgac gtcgacggat cgggagatct cccgatcccc tatggtgcac tctcagtaca
     7861 atctgctctg atgccgcata gttaagccag tatctgctc
//
