LOCUS       sequence-218380-        7755 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of human PLAA with C-terminal strep-HA tag.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7755)
  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 7755)
  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-PLAA-Strep-HA
FEATURES             Location/Qualifiers
     source          1..7755
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        172..551
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        552..755
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     706..726
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     protein_bind    757..775
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    778..796
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     806..830
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     protein_bind    973..997
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(3389..3413)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             3460..3483
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     CDS             3520..3543
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     CDS             3574..3600
                     /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             3574..3600
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     primer_bind     complement(3644..3661)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3650..3874
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(4007..4026)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     protein_bind    4158..4205
                     /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             4213..5235
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="KKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGY
                     VLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPE
                     TELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQT
                     VMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFG
                     DSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGN
                     FDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE
                     "
     CDS             4213..5235
                     /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    5365..5486
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5402..5421)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5456..5475
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5535..5551)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5535..5551)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5548..5570)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5559..5575
                     /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(5583..5589,5590..5607,5608..5613))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5628..5649
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5766..5783)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5937..6525)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6017..6036)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6696..7487,7488..7556))
                     /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     7319..7338
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7557..7661)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(7736..7755)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 catagttaag ccagtatctg ctccctgctt gtgtgttgga ggtcgctgag tagtgcgcga
       61 gcaaaattta agctacaaca aggcaaggct tgaccgacaa ttgcatgaag aatctgctta
      121 gggttaggcg ttttgcgctg cttcgcgatg tacgggccag atatacgcgt tgacattgat
      181 tattgactag ttattaatag taatcaatta cggggtcatt agttcatagc ccatatatgg
      241 agttccgcgt tacataactt acggtaaatg gcccgcctgg ctgaccgccc aacgaccccc
      301 gcccattgac gtcaataatg acgtatgttc ccatagtaac gccaataggg actttccatt
      361 gacgtcaatg ggtggagtat ttacggtaaa ctgcccactt ggcagtacat caagtgtatc
      421 atatgccaag tacgccccct attgacgtca atgacggtaa atggcccgcc tggcattatg
      481 cccagtacat gaccttatgg gactttccta cttggcagta catctacgta ttagtcatcg
      541 ctattaccat ggtgatgcgg ttttggcagt acatcaatgg gcgtggatag cggtttgact
      601 cacggggatt tccaagtctc caccccattg acgtcaatgg gagtttgttt tggcaccaaa
      661 atcaacggga ctttccaaaa tgtcgtaaca actccgcccc attgacgcaa atgggcggta
      721 ggcgtgtacg gtgggaggtc tatataagca gagctctccc tatcagtgat agagatctcc
      781 ctatcagtga tagagatcgt cgacgagctc gtttagtgaa ccgtcagatc gcctggagac
      841 gccatccacg ctgttttgac ctccatagaa gacaccggga ccgatccagc ctccggactc
      901 tagcgtttaa acttaagctt ggtaccgagc tcggatccac tagtccagtg tggtggaatt
      961 ctgcagatat caacaagttt gtacaaaaaa gcaggctcca ccatgacgag cggcgcaacc
     1021 aggtaccggc tgagctgctc gctccggggc cacgagctgg acgtacgggg cctggtgtgc
     1081 tgcgcctatc cgccgggagc ctttgtgtcc gtgtcccgag accgcaccac ccgcctctgg
     1141 gccccagaca gtccaaacag gagctttaca gaaatgcact gtatgagtgg ccattccaat
     1201 tttgtatctt gtgtatgcat cataccctca agtgacatct accctcatgg cctaattgcc
     1261 accggtggaa atgaccacaa tatatgcatt ttctcactgg acagtccaat gccactttat
     1321 attctaaaag gccacaaaaa tactgtttgt agtctatcat ctggaaaatt tgggacatta
     1381 cttagtggtt catgggacac cactgctaaa gtctggctga atgacaagtg catgatgacc
     1441 ttgcagggtc atacagctgc agtgtgggcg gtaaagatct tacctgaaca gggcttaatg
     1501 ttgactggat cagcagacaa gactgttaaa ctgtggaagg ctggaagatg tgagaggact
     1561 ttttcagggc atgaagactg tgtaagaggt ttggcaattt tgagtgaaac agaatttctt
     1621 tcctgtgcaa atgatgctag tattagaagg tggcaaatca ctggcgagtg tcttgaagta
     1681 tattatggac atacaaatta tatttatagc atatccgttt ttccaaattg tagagacttt
     1741 gtgacaacag cagaggacag atctctgaga atctggaaac atggggaatg tgctcaaact
     1801 atccgacttc cagctcagtc tatatggtgc tgctgtgtgc tcgacaatgg tgacattgtg
     1861 gttggtgcga gtgatggcat tattagagtg tttacagaat cagaagatcg aacagcaagt
     1921 gctgaagaaa tcaaggcttt tgaaaaagaa ctgtctcacg caaccattga ttctaaaact
     1981 ggcgatttag gggacatcaa tgctgagcag cttcctggga gggaacatct taatgaacct
     2041 ggtactagag aaggacagac tcgtctaatc agagatgggg agaaagtcga agcctatcag
     2101 tggagtgtta gtgaagggag gtggataaaa attggtgatg ttgttggctc atctggtgct
     2161 aatcagcaaa catctggaaa agttttatat gaagggaaag aatttgatta tgttttctca
     2221 attgatgtca atgaaggtgg accatcatat aaattgccat ataataccag tgatgaccct
     2281 tggttaactg catacaactt cttacagaag aatgatttga atcctatgtt tctggatcaa
     2341 gtagctaaat ttattattga taacacaaaa ggtcaaatgt tgggacttgg gaatcccagc
     2401 ttttcagatc catttacagg tggtggtcgg tatgttccgg gctcttcggg atcttctaac
     2461 acactaccca cagcagatcc ttttacaggt gctggtcgtt atgtaccagg ttctgcaagt
     2521 atgggaacta ccatggccgg agttgatcca tttacaggga atagtgccta ccgatcagct
     2581 gcatctaaaa caatgaatat ttatttccct aaaaaagagg ctgtcacatt tgaccaagca
     2641 aaccctacac aaatattagg taaactgaag gaacttaatg gaactgcacc tgaagagaag
     2701 aagttaactg aggatgactt gatacttctt gagaagatac tgtctctaat atgtaatagt
     2761 tcttcagaaa aacccacagt ccagcaactt cagattttgt ggaaagctat taactgtcct
     2821 gaagatattg tctttcctgc acttgacatt cttcggttgt caattaaaca ccccagtgtg
     2881 aatgagaact tctgcaatga aaaggaaggg gctcagttca gcagtcatct tatcaatctt
     2941 ctgaacccta aaggaaagcc agcaaaccag ctgcttgctc tcaggacttt ttgcaattgt
     3001 tttgttggcc aggcaggaca aaaactcatg atgtcccaga gggaatcact gatgtcccat
     3061 gcaatagaac tgaaatcagg gagcaataag aacattcaca ttgctctggc tacattggcc
     3121 ctgaactatt ctgtttgttt tcataaagac cataacattg aagggaaagc ccaatgtttg
     3181 tcactaatta gcacaatctt ggaagtagta caagacctag aagccacttt tagacttctt
     3241 gtggctcttg gaacacttat cagtgatgat tcaaatgctg tacaattagc caagtcttta
     3301 ggtgttgatt ctcaaataaa aaagtattcc tcagtatcag aaccagctaa agtaagtgaa
     3361 tgctgtagat ttatcctaaa tttgctgtac ccagctttct tgtacaaagt ggttgatatc
     3421 aaggccgccg acatcaccag cctgtacaag aaagtggggt ggagccatcc ccagttcgag
     3481 aagggcggag gatctggcgg cggaagcggc ggaggaagct ggtcccaccc tcagtttgag
     3541 aaaggcaccg agctgggcag caccatggcc agctacccct acgacgtgcc cgactacgcc
     3601 tgatgactcg agtctagagg gcccgtttaa acccgctgat cagcctcgac tgtgccttct
     3661 agttgccagc catctgttgt ttgcccctcc cccgtgcctt ccttgaccct ggaaggtgcc
     3721 actcccactg tcctttccta ataaaatgag gaaattgcat cgcattgtct gagtaggtgt
     3781 cattctattc tggggggtgg ggtggggcag gacagcaagg gggaggattg ggaagacaat
     3841 agcaggcatg ctggggatgc ggtgggctct atggcttctg aggcggaaag aaccagctgg
     3901 ggctctaggg ggtatcccca cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg
     3961 gttacgcgca gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc tttcgctttc
     4021 ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcgggggctc
     4081 cctttagggt tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgattagggt
     4141 gatggttcac gtacctagaa gttcctattc cgaagttcct attctctaga aagtatagga
     4201 acttccttgg ccaaaaagcc tgaactcacc gcgacgtctg tcgagaagtt tctgatcgaa
     4261 aagttcgaca gcgtctccga cctgatgcag ctctcggagg gcgaagaatc tcgtgctttc
     4321 agcttcgatg taggagggcg tggatatgtc ctgcgggtaa atagctgcgc cgatggtttc
     4381 tacaaagatc gttatgttta tcggcacttt gcatcggccg cgctcccgat tccggaagtg
     4441 cttgacattg gggaattcag cgagagcctg acctattgca tctcccgccg tgcacagggt
     4501 gtcacgttgc aagacctgcc tgaaaccgaa ctgcccgctg ttctgcagcc ggtcgcggag
     4561 gccatggatg cgatcgctgc ggccgatctt agccagacga gcgggttcgg cccattcgga
     4621 ccgcaaggaa tcggtcaata cactacatgg cgtgatttca tatgcgcgat tgctgatccc
     4681 catgtgtatc actggcaaac tgtgatggac gacaccgtca gtgcgtccgt cgcgcaggct
     4741 ctcgatgagc tgatgctttg ggccgaggac tgccccgaag tccggcacct cgtgcacgcg
     4801 gatttcggct ccaacaatgt cctgacggac aatggccgca taacagcggt cattgactgg
     4861 agcgaggcga tgttcgggga ttcccaatac gaggtcgcca acatcttctt ctggaggccg
     4921 tggttggctt gtatggagca gcagacgcgc tacttcgagc ggaggcatcc ggagcttgca
     4981 ggatcgccgc ggctccgggc gtatatgctc cgcattggtc ttgaccaact ctatcagagc
     5041 ttggttgacg gcaatttcga tgatgcagct tgggcgcagg gtcgatgcga cgcaatcgtc
     5101 cgatccggag ccgggactgt cgggcgtaca caaatcgccc gcagaagcgc ggccgtctgg
     5161 accgatggct gtgtagaagt actcgccgat agtggaaacc gacgccccag cactcgtccg
     5221 agggcaaagg aatagcacgt actacgagat ttcgattcca ccgccgcctt ctatgaaagg
     5281 ttgggcttcg gaatcgtttt ccgggacgcc ggctggatga tcctccagcg cggggatctc
     5341 atgctggagt tcttcgccca ccccaacttg tttattgcag cttataatgg ttacaaataa
     5401 agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc tagttgtggt
     5461 ttgtccaaac tcatcaatgt atcttatcat gtctgtatac cgtcgacctc tagctagagc
     5521 ttggcgtaat catggtcata gctgtttcct gtgtgaaatt gttatccgct cacaattcca
     5581 cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg agtgagctaa
     5641 ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag
     5701 ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc
     5761 gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct
     5821 cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg
     5881 tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc
     5941 cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga
     6001 aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct
     6061 cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg
     6121 gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag
     6181 ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat
     6241 cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac
     6301 aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac
     6361 tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc
     6421 ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt
     6481 tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc
     6541 ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg
     6601 agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca
     6661 atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca
     6721 cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag
     6781 ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac
     6841 ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc
     6901 agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct
     6961 agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc
     7021 gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg
     7081 cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc
     7141 gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat
     7201 tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag
     7261 tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat
     7321 aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg
     7381 cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca
     7441 cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga
     7501 aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc
     7561 ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag cggatacata
     7621 tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg
     7681 ccacctgacg tcgacggatc gggagatctc ccgatcccct atggtgcact ctcagtacaa
     7741 tctgctctga tgccg
//
