LOCUS       sequence-270198-        8153 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses full-length Protein disulfide-isomerase A3 precursor 
            ectodomain in mammalian cells. C-terminal rat Cd4d3+4 tag, 
            biotinylation sequence and His tag. .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8153)
  AUTHORS   Sun Y, Vandenbriele C, Kauskot A, Verhamme P, Hoylaerts MF, Wright 
            GJ
  TITLE     A human platelet receptor protein microarray identifies 
            FcepsilonR1alpha as an activating PEAR1 ligand.
  JOURNAL   Mol Cell Proteomics. 2015 Feb 23. pii: mcp.M114.046946.
  PUBMED    25713122
REFERENCE   2  (bases 1 to 8153)
  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. 2015 Feb 23. pii: mcp.M114.046946."
COMMENT     Sequence Label: PDIA3-bio-His
FEATURES             Location/Qualifiers
     source          1..8153
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..380
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        381..584
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     535..555
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     581..605
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     primer_bind     1086..1105
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     regulatory      1139..1148
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3311..3328
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    3380..3435
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(3434..3453)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(5671..5689)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5757..5861
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5862..5930,5931..6722)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(6080..6099)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6893..7481
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7382..7401
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7635..7652
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    7769..7790
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7805..7810,7811..7828,7829..7835)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7843..7859
                     /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)."
     primer_bind     7848..7870
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7867..7883
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7867..7883
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc
       61 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
      121 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
      181 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
      241 aagtgtatca tatgccaagt ccgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
      301 ggcattatgc ccagtacatg accttacggg actttcctac ttggcagtac atctacgtat
      361 tagtcatcgc tattaccatg gtgatgcggt tttggcagta caccaatggg cgtggatagc
      421 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt
      481 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaataa ccccgccccg ttgacgcaaa
      541 tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctcgttta gtgaaccgtc
      601 agatcctcac tctcttccgc atcgctgtct gcgagggcca gctgttgggc tcgcggttga
      661 ggacaaactc ttcgcggtct ttccagtact cttggatcgg aaacccgtcg gcctccgaac
      721 ggtactccgc caccgaggga cctgagcgag tccgcatcga ccggatcgga aaacctctcg
      781 agaaaggcgt ctaaccagtc acagtcgcaa ggtaggctga gcaccgtggc gggcggcagc
      841 gggtggcggt cggggttgtt tctggcggag gtgctgctga tgatgtaatt aaagtaggcg
      901 gtcttgagac ggcggatggt cgaggtgagg tgtggcaggc ttgagatcca gctgttgggg
      961 tgagtactcc ctctcaaaag cgggcattac ttctgcgcta agattgtcag tttccaaaaa
     1021 cgaggaggat ttgatattca cctggcccga tctggccata cacttgagtg acaatgacat
     1081 ccactttgcc tttctctcca caggtgtcca ctcccaggtc caagtttaaa ctgcggccgc
     1141 caccatggct ctgcggagac tggccctgtt tcctggcgtg gccctgctgc tggctgctgc
     1201 tagactggcc gctgccagcg acgtgctgga actgaccgac gacaacttcg agagccggat
     1261 cagcgatacc ggcagcgccg gactgatgct ggtcgagttc ttcgcccctt ggtgcggcca
     1321 ctgcaagagg ctggcccctg agtacgaggc tgccgccacc agactgaagg gcatcgtgcc
     1381 cctggccaag gtggactgca ccgccaacac caacacctgt aacaagtacg gcgtgagcgg
     1441 ctaccccacc ctgaagatct tccgggacgg cgaggaagct ggcgcctacg acggccctag
     1501 aaccgccgat ggcatcgtgt cccacctgaa gaagcaggcc ggaccagcca gcgtgcccct
     1561 gcggaccgag gaagagttca agaagttcat cagcgacaag gacgccagca tcgtgggctt
     1621 tttcgacgac agcttcagcg aggcccacag cgagtttctg aaggccgcca gcaacctgag
     1681 agacaactac agattcgccc acaccaacgt ggagagcctg gtcaacgagt acgacgacaa
     1741 cggcgagggc atcatcctgt tccggcccag ccacctgacc aacaagttcg aggacaagac
     1801 cgtggcctac accgagcaga agatgaccag cggcaagatc aagaagttta tccaggaaaa
     1861 catcttcggc atctgccccc acatgaccga ggacaacaag gacctgatcc agggcaagga
     1921 tctgctgatc gcctactacg acgtggacta cgagaagaac gccaagggca gcaactactg
     1981 gcggaacaga gtgatgatgg tggccaagaa gttcctggac gccggccaca agctgaactt
     2041 cgccgtggcc agcagaaaga ccttcagcca cgagctgtcc gacttcggcc tggaaagcac
     2101 agccggcgag atccccgtgg tggccatcag aaccgccaag ggggagaagt tcgtgatgca
     2161 ggaagagttt agccgggacg gcaaggccct ggaacggttc ctgcaggact acttcgacgg
     2221 caacctgaag agatacctga agtccgagcc catccccgag agcaacgacg gccccgtgaa
     2281 ggtggtggtg gccgagaact tcgacgagat cgtgaacaac gagaacaagg acgtgctgat
     2341 cgaattctat gctccttggt gtgggcattg caagaacctg gaacccaagt acaaagagct
     2401 gggcgagaag ctgtccaagg accccaatat cgtgatcgcc aagatggacg ccacagccaa
     2461 cgacgtgccc agcccttacg aagtgcgcgg cttccccacc atctacttca gccccgccaa
     2521 caagaagctg aaccccaaga agtacgaggg cggcagagaa ctgagcgact tcatcagcta
     2581 tctgcagcgg gaggccacca atccccctgt gatccaggaa gaaaagccca agaagaagaa
     2641 aaaggcccag gaagatctcg gcgcgccgtc gacctccatc acggcctata agagtgaggg
     2701 ggagtcagcg gagttctcct tcccactcaa ccttggagag gaaagcctgc agggagagtt
     2761 gagatggaag gcagagaagg ctccttcttc ccagtcctgg atcaccttct ccctaaagaa
     2821 ccaaaaggtg tctgtgcaga agtctactag caaccccaag ttccagctgt ccgaaacgct
     2881 cccactcacc cttcagatac cccaggtctc ccttcagttt gctggttctg gcaacctgac
     2941 cctgactctg gacagaggga tactgtatca ggaagtgaac ctggtggtga tgaaagtgac
     3001 tcagcccgac agcaacactt tgacctgtga ggtgatggga cccacctcac ccaagatgag
     3061 actgatcttg aagcaggaga atcaggaggc cagggtctcc aggcaggaga aagtgattca
     3121 agtgcaggcc cctgaagcag gggtgtggca atgtctactg agtgaaggtg aagaggtcaa
     3181 gatggactcc aagatccagg ttttatccaa agggttgaat tccggcagcc tgcaccacat
     3241 cctggacgcc cagaagatgc tgtggaacca cagggacagg aacctgcctc ctctggcccc
     3301 tctgggcccc caccaccatc accaccattg agtctagaca cgtgattaat taaggatccc
     3361 ccgacctcga cctctggcta ataaaggaaa tttattttca ttgcaatagt gtgttggaat
     3421 tttttgtgtc tctcactcgg aaggacatat gggagggcaa atcatttggt cgagatccct
     3481 cggagatctc tagctagagg atcgatcccc gccccggacg aactaaacct gactacgaca
     3541 tctctgcccc ttcttcgcgg ggcagtgcat gtaatccctt cagttggttg gtacaacttg
     3601 ccaactgggc cctgttccac atgtgacacg gggggggacc aaacacaaag gggttctctg
     3661 actgtagttg acatccttat aaatggatgt gcacatttgc caacactgag tggctttcat
     3721 cctggagcag actttgcagt ctgtggactg caacacaaca ttgcctttat gtgtaactct
     3781 tggctgaagc tcttacacca atgctggggg acatgtacct cccaggggcc caggaagact
     3841 acgggaggct acaccaacgt caatcagagg ggcctgtgta gctaccgata agcggaccct
     3901 caagagggca ttagcaatag tgtttataag gcccccttgt taaccctaaa cgggtagcat
     3961 atgcttcccg ggtagtagta tatactatcc agactaaccc taattcaata gcatatgtta
     4021 cccaacggga agcatatgct atcgaattag ggttagtaaa agggtcctaa ggaacagcga
     4081 tatctcccac cccatgagct gtcacggttt tatttacatg gggtcaggat tccacgaggg
     4141 tagtgaacca ttttagtcac aagggcagtg gctgaagatc aaggagcggg cagtgaactc
     4201 tcctgaatct tcgcctgctt cttcattctc cttcgtttag ctaatagaat aactgctgag
     4261 ttgtgaacag taaggtgtat gtgaggtgct cgaaaacaag gtttcaggtg acgcccccag
     4321 aataaaattt ggacgggggg ttcagtggtg gcattgtgct atgacaccaa tataaccctc
     4381 acaaacccct tgggcaataa atactagtgt aggaatgaaa cattctgaat atctttaaca
     4441 atagaaatcc atggggtggg gacaagccgt aaagactgga tgtccatctc acacgaattt
     4501 atggctatgg gcaacacata atcctagtgc aatatgatac tggggttatt aagatgtgtc
     4561 ccaggcaggg accaagacag gtgaaccatg ttgttacact ctatttgtaa caaggggaaa
     4621 gagagtggac gccgacagca gcggactcca ctggttgtct ctaacacccc cgaaaattaa
     4681 acggggctcc acgccaatgg ggcccataaa caaagacaag tggccactct tttttttgaa
     4741 attgtggagt gggggcacgc gtcagccccc acacgccgcc ctgcggtttt ggactgtaaa
     4801 ataagggtgt aataacttgg ctgattgtaa ccccgctaac cactgcggtc aaaccacttg
     4861 cccacaaaac cactaatggc accccgggga atacctgcat aagtaggtgg gcgggccaag
     4921 ataggggcgc gattgctgcg atctggagga caaattacac acacttgcgc ctgagcgcca
     4981 agcacagggt tgttggtcct catattcacg aggtcgctga gagcacggtg ggctaatgtt
     5041 gccatgggta gcatatacta cccaaatatc tggatagcat atgctatcct aatctatatc
     5101 tgggtagcat aggctatcct aatctatatc tgggtagcat atgctatcct aatctatatc
     5161 tgggtagtat atgctatcct aatttatatc tgggtagcat aggctatcct aatctatatc
     5221 tgggtagcat atgctatcct aatctatatc tgggtagtat atgctatcct aatctgtatc
     5281 cgggtagcat atgctatcct aatagagatt agggtagtat atgctatcct aatttatatc
     5341 tgggtagcat atactaccca aatatctgga tagcatatgc tatcctaatc tatatctggg
     5401 tagcatatgc tatcctaatc tatatctggg tagcataggc tatcctaatc tatatctggg
     5461 tagcatatgc tatcctaatc tatatctggg tagtatatgc tatcctaatt tatatctggg
     5521 tagcataggc tatcctaatc tatatctggg tagcatatgc tatcctaatc tatatctggg
     5581 tagtatatgc tatcctaatc tgtatccggg tagcatatgc tatcctcatg ataagctgtc
     5641 aaacatgaga attttcttga agacgaaagg gcctcgtgat acgcctattt ttataggtta
     5701 atgtcatgat aataatggtt tcttagacgt caggtggcac ttttcgggga aatgtgcgcg
     5761 gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat
     5821 aaccctgata aatgcttcaa taatattgaa aaaggaagag tatgagtatt caacatttcc
     5881 gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct cacccagaaa
     5941 cgctggtgaa agtaaaagat gctgaagatc agttgggtgc acgagtgggt tacatcgaac
     6001 tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgt tttccaatga
     6061 tgagcacttt taaagttctg ctatgtggcg cggtattatc ccgtgttgac gccgggcaag
     6121 agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtac tcaccagtca
     6181 cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct gccataacca
     6241 tgagtgataa cactgcggcc aacttacttc tgacaacgat cggaggaccg aaggagctaa
     6301 ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg gaaccggagc
     6361 tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgcagca atggcaacaa
     6421 cgttgcgcaa actattaact ggcgaactac ttactctagc ttcccggcaa caattaatag
     6481 actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt ccggctggct
     6541 ggtttattgc tgataaatct ggagccggtg agcgtgggtc tcgcggtatc attgcagcac
     6601 tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg agtcaggcaa
     6661 ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt aagcattggt
     6721 aactgtcaga ccaagtttac tcatatatac tttagattga tttaaaactt catttttaat
     6781 ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg
     6841 agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc
     6901 ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg
     6961 tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag
     7021 cgcagatacc aaatactgtt cttctagtgt agccgtagtt aggccaccac ttcaagaact
     7081 ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg
     7141 gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc
     7201 ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg
     7261 aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg
     7321 cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag
     7381 ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc
     7441 gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct
     7501 ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct gcgttatccc
     7561 ctgattctgt ggataaccgt attaccgcct ttgagtgagc tgataccgct cgccgcagcc
     7621 gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga agagcgccca atacgcaaac
     7681 cgcctctccc cgcgcgttgg ccgattcatt aatgcagctg gcacgacagg tttcccgact
     7741 ggaaagcggg cagtgagcgc aacgcaatta atgtgagtta gctcactcat taggcacccc
     7801 aggctttaca ctttatgctt ccggctcgta tgttgtgtgg aattgtgagc ggataacaat
     7861 ttcacacagg aaacagctat gaccatgatt acgccaagct ctagctagag gtcgaccaat
     7921 tctcatgttt gacagcttat catcgcagat ccgggcaacg ttgttgccat tgctgcaggc
     7981 gcagaactgg taggtatgga agatctatac attgaatcaa tattggcaat tagccatatt
     8041 agtcattggt tatatagcat aaatcaatat tggctattgg ccattgcata cgttgtatct
     8101 atatcataat atgtacattt atattggctc atgtccaata tgaccgccat gtt
//
