LOCUS       sequence-208819-        9679 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses pentamerised full-length protein ectodomain with no 
            N-linked glycans in mammalian cells. Contains a C-terminal rat Cd4 
            d3+4 tag, COMP pentamerisation domain, flag- and 6xHis tags. .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9679)
  AUTHORS   Knockel J, Dundas K, Yang ASP, Galaway F, Metcalf T, Gemert GV, 
            Sauerwein RW, Rayner JC, Billker O, Wright GJ
  TITLE     Systematic Identification of Plasmodium Falciparum Sporozoite 
            Membrane Protein Interactions Reveals an Essential Role for the p24 
            Complex in Host Infection.
  JOURNAL   Mol Cell Proteomics. 2021;20:100038. doi: 10.1074/mcp.RA120.002432. 
            Epub 2021 Jan 27.
  PUBMED    33515807
REFERENCE   2  (bases 1 to 9679)
  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."; date: "2021"; volume: "20:100038. doi"; pages: " 
            10.1074/mcp.RA120.002432. Epub 2021 Jan 27"
COMMENT     Sequence Label: DPAP1-COMP-blac-flag-his
FEATURES             Location/Qualifiers
     source          1..9679
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     310..329
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     regulatory      352..361
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3211..3996
                     /codon_start=1
                     /gene="bla(M)"
                     /product="beta-lactamase lacking the signal sequence"
                     /label=bla(M)
                     /note="allows cytosolic expression of beta-lactamase"
                     /translation="PETLVKVKDAEDQLGARVGYIELDLNSGKILESFRPEERFPMMST
                     FKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYSPVTEKHLTDGMTVRELCSAAITMS
                     DNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRWEPELNEAIPNDERDTTMPVAMATT
                     LRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSALPAGWFIADKSGAGERGSRGIIAA
                     LGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGASLIKHW"
     CDS             4003..4068
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             4075..4092
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    4130..4185
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(4184..4203)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(6421..6439)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6507..6611
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6612..6680,6681..7472)
                     /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"
     rep_origin      7643..8231
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8132..8151
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     8385..8402
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    8519..8540
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(8555..8560,8561..8578,8579..8585)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8593..8609
                     /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     8598..8620
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     8617..8633
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     8617..8633
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     enhancer        8904..9283
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        9284..9487
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     9438..9458
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     9484..9508
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
ORIGIN
        1 ctcgagaaag gcgtctaacc agtcacagtc gcaaggtagg ctgagcaccg tggcgggcgg
       61 cagcgggtgg cggtcggggt tgtttctggc ggaggtgctg ctgatgatgt aattaaagta
      121 ggcggtcttg agacggcgga tggtcgaggt gaggtgtggc aggcttgaga tccagctgtt
      181 ggggtgagta ctccctctca aaagcgggca ttacttctgc gctaagattg tcagtttcca
      241 aaaacgagga ggatttgata ttcacctggc ccgatctggc catacacttg agtgacaatg
      301 acatccactt tgcctttctc tccacaggtg tccactccca ggtccaagtt tgccaccatg
      361 gagtttcaga cccaggtact catgtccctg ctgctctgca tgtctggtgc ggccgccgat
      421 ctgcccaccc acgtggaaac aaagaacctg ctgggcaagt ggaagatcct gcggaccaag
      481 accagcccca acctggccac ctgtggcagc agccagccca acaagaacac ctacaacgtg
      541 ggcatcaccg actacaagaa gtacctgctg gaaaacaact acgagttcgt gtccgagctg
      601 aacgtgatcc tgagcgacga ctacgtgctg tacggcgaca tctacaacac ccaggacaac
      661 gagcaccggt ctaagtggaa ggtgctggcc gtgtacgacg agaacaagag agtgatcggc
      721 acctggacca ccatctgcga cgagggcttc gagatcaaga tcggcaacga ggcctacgcc
      781 gccctgatgc actacgagcc caacggcaag tgcggccctg tgtccgacga ggacagcctg
      841 gacagcaacg gcgacaccga ctgctacacc accagcttca gcaagatcag atacggctgg
      901 ctggacgtgg aaaacgagaa gaacgagcat ctgcacggct gcttctacgc cgagcggatc
      961 ttcgacaacg tgaacgagat taagcacctc gacagcttca ccatcgacaa ggactcccag
     1021 aacgtgctgc agaccttcac ctacgacacc aagctgaaca acatcctgaa cagcaacaac
     1081 atgctgtaca agttcggcaa cctgcagaag cccaccttca ccaagagaaa caacgccaac
     1141 gtgcagttca acagcgagct gaattggcac cggatgaagc accacggcaa gaagaagccc
     1201 ctgaaaaaga gcatgctgga cgccagccgg cagacctacg cctgcccctg caacgccaat
     1261 gaggtggtgg acaatgtgat caacaagggc gacagcgaca accccgtgtc ccctaccctg
     1321 atccagctca acaacaatct gaagaacacc gcccagaccg gcaacaagga caccaacgag
     1381 atggacctgg aaaattacga ggacacactg aacagcccca agcgcgagct ggaaatcaac
     1441 gagctgccca agaacttcgc ctggggcgac ccctggaaca agaatacccg cgagtacgaa
     1501 gtgaccaacc agctgctgtg cggcagctgc tatatcgcca gccagctgta cgccttcaag
     1561 cggcggatcg aggtggccct gaccaagaag ctggacagaa agtacctgaa caacttcgac
     1621 gaccagctga gcatccagac cgtgctgtcc tgcagcttct acgaccaggg ctgcaacggc
     1681 ggcttcccct acctggtgtc caagctggcc aaactgcagg gcatccctct gaacgtgtac
     1741 ttcccttaca gcgccaccga ggaaacctgc ccctacaata tcgccaagca ccccaacgac
     1801 atgaacggcg ctgccaagct gagagagatc aacgccatct tcaactctaa caacaatatg
     1861 gccacataca acaacatcaa caacgaccat caccagctgg gagtgtacgc caacaccgcc
     1921 agctcccagg aacagcacgg catcagcgaa gagaaccggt ggtacgccaa ggacttcaac
     1981 tacgtgggcg gctgctacgg ctgcaaccag tgcaatggcg agaagatcat gatgaatgag
     2041 atctaccgga acggccccat cgtgtctagc ttcgaggcca gccccgactt ctacgattac
     2101 gccgacggcg tgtactttgt ggaagatttc cctcacgcca gacggtgcac catcgagccc
     2161 aagaatgatg gcgtgtacaa cattgccgga tgggacagag tgaaccacgc catcgtgctg
     2221 ctgggctggg gcgaggaaga gattaacggc aagctgtata agtactggat cggccggaac
     2281 agctggggca acggctgggg aaaagagggc tacttcaaga tcctgagagg ccagaatttc
     2341 gccggcatcg agagccagag cctgttcatc gagcctgact tcagccgggg agccggcaag
     2401 attctgctcg agaagatgca gaaagagctg ggaaacggcg cgccgtcgac ctccatcacg
     2461 gcctataaga gtgaggggga gtcagcggag ttctccttcc cactcaacct tggagaggaa
     2521 agcctgcagg gagagttgag atggaaggca gagaaggctc cttcttccca gtcctggatc
     2581 accttctccc taaagaacca aaaggtgtct gtgcagaagt ctactagcaa ccccaagttc
     2641 cagctgtccg aaacgctccc actcaccctt cagatacccc aggtctccct tcagtttgct
     2701 ggttctggca acctgaccct gactctggac agagggatac tgtatcagga agtgaacctg
     2761 gtggtgatga aagtgactca gcccgacagc aacactttga cctgtgaggt gatgggaccc
     2821 acctcaccca agatgagact gatcttgaag caggagaatc aggaggccag ggtctccagg
     2881 caggagaaag tgattcaagt gcaggcccct gaagcagggg tgtggcaatg tctactgagt
     2941 gaaggtgaag aggtcaagat ggactccaag atccaggttt tatccaaagg gttgaattcc
     3001 ggcggcggct ccggaggcgg taccggtggc ggcgacctag ccccacagat gcttcgagaa
     3061 ctccaggaga ctaatgcggc gctgcaagac gtgagagagc tcttgcgaca gcaggtcaag
     3121 gagatcacct tcctgaagaa tacggtgatg gaatgtgacg cctgcggtct agacagaaac
     3181 ctgcctccac tagcacctct gggcccacac ccagaaacgc tggtgaaagt aaaagatgct
     3241 gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag cggtaagatc
     3301 cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa agttctgcta
     3361 tgtggcgcgg tattatcccg tattgacgcc gggcaagagc aactcggtcg ccgcatacac
     3421 tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct tacggatggc
     3481 atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac
     3541 ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca caacatgggg
     3601 gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat accaaacgac
     3661 gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact attaactggc
     3721 gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc ggataaagtt
     3781 gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga taaatctgga
     3841 gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg taagccctcc
     3901 cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag
     3961 atcgctgaga taggtgcctc actgattaag cattggggcc ctgactacaa agaccatgac
     4021 ggtgattata aagatcatga catcgattac aaggatgacg atgacaaggg ccctcaccac
     4081 caccatcacc actaattaat taaggatccc ccgacctcga cctctggcta ataaaggaaa
     4141 tttattttca ttgcaatagt gtgttggaat tttttgtgtc tctcactcgg aaggacatat
     4201 gggagggcaa atcatttggt cgagatccct cggagatctc tagctagagg atcgatcccc
     4261 gccccggacg aactaaacct gactacgaca tctctgcccc ttcttcgcgg ggcagtgcat
     4321 gtaatccctt cagttggttg gtacaacttg ccaactgggc cctgttccac atgtgacacg
     4381 gggggggacc aaacacaaag gggttctctg actgtagttg acatccttat aaatggatgt
     4441 gcacatttgc caacactgag tggctttcat cctggagcag actttgcagt ctgtggactg
     4501 caacacaaca ttgcctttat gtgtaactct tggctgaagc tcttacacca atgctggggg
     4561 acatgtacct cccaggggcc caggaagact acgggaggct acaccaacgt caatcagagg
     4621 ggcctgtgta gctaccgata agcggaccct caagagggca ttagcaatag tgtttataag
     4681 gcccccttgt taaccctaaa cgggtagcat atgcttcccg ggtagtagta tatactatcc
     4741 agactaaccc taattcaata gcatatgtta cccaacggga agcatatgct atcgaattag
     4801 ggttagtaaa agggtcctaa ggaacagcga tatctcccac cccatgagct gtcacggttt
     4861 tatttacatg gggtcaggat tccacgaggg tagtgaacca ttttagtcac aagggcagtg
     4921 gctgaagatc aaggagcggg cagtgaactc tcctgaatct tcgcctgctt cttcattctc
     4981 cttcgtttag ctaatagaat aactgctgag ttgtgaacag taaggtgtat gtgaggtgct
     5041 cgaaaacaag gtttcaggtg acgcccccag aataaaattt ggacgggggg ttcagtggtg
     5101 gcattgtgct atgacaccaa tataaccctc acaaacccct tgggcaataa atactagtgt
     5161 aggaatgaaa cattctgaat atctttaaca atagaaatcc atggggtggg gacaagccgt
     5221 aaagactgga tgtccatctc acacgaattt atggctatgg gcaacacata atcctagtgc
     5281 aatatgatac tggggttatt aagatgtgtc ccaggcaggg accaagacag gtgaaccatg
     5341 ttgttacact ctatttgtaa caaggggaaa gagagtggac gccgacagca gcggactcca
     5401 ctggttgtct ctaacacccc cgaaaattaa acggggctcc acgccaatgg ggcccataaa
     5461 caaagacaag tggccactct tttttttgaa attgtggagt gggggcacgc gtcagccccc
     5521 acacgccgcc ctgcggtttt ggactgtaaa ataagggtgt aataacttgg ctgattgtaa
     5581 ccccgctaac cactgcggtc aaaccacttg cccacaaaac cactaatggc accccgggga
     5641 atacctgcat aagtaggtgg gcgggccaag ataggggcgc gattgctgcg atctggagga
     5701 caaattacac acacttgcgc ctgagcgcca agcacagggt tgttggtcct catattcacg
     5761 aggtcgctga gagcacggtg ggctaatgtt gccatgggta gcatatacta cccaaatatc
     5821 tggatagcat atgctatcct aatctatatc tgggtagcat aggctatcct aatctatatc
     5881 tgggtagcat atgctatcct aatctatatc tgggtagtat atgctatcct aatttatatc
     5941 tgggtagcat aggctatcct aatctatatc tgggtagcat atgctatcct aatctatatc
     6001 tgggtagtat atgctatcct aatctgtatc cgggtagcat atgctatcct aatagagatt
     6061 agggtagtat atgctatcct aatttatatc tgggtagcat atactaccca aatatctgga
     6121 tagcatatgc tatcctaatc tatatctggg tagcatatgc tatcctaatc tatatctggg
     6181 tagcataggc tatcctaatc tatatctggg tagcatatgc tatcctaatc tatatctggg
     6241 tagtatatgc tatcctaatt tatatctggg tagcataggc tatcctaatc tatatctggg
     6301 tagcatatgc tatcctaatc tatatctggg tagtatatgc tatcctaatc tgtatccggg
     6361 tagcatatgc tatcctcatg ataagctgtc aaacatgaga attttcttga agacgaaagg
     6421 gcctcgtgat acgcctattt ttataggtta atgtcatgat aataatggtt tcttagacgt
     6481 caggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac
     6541 attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa
     6601 aaaggaagag tatgagtatt caacatttcc gtgtcgccct tattcccttt tttgcggcat
     6661 tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat gctgaagatc
     6721 agttgggtgc acgagtgggt tacatcgaac tggatctcaa cagcggtaag atccttgaga
     6781 gttttcgccc cgaagaacgt tttccaatga tgagcacttt taaagttctg ctatgtggcg
     6841 cggtattatc ccgtgttgac gccgggcaag agcaactcgg tcgccgcata cactattctc
     6901 agaatgactt ggttgagtac tcaccagtca cagaaaagca tcttacggat ggcatgacag
     6961 taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc aacttacttc
     7021 tgacaacgat cggaggaccg aaggagctaa ccgctttttt gcacaacatg ggggatcatg
     7081 taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac gacgagcgtg
     7141 acaccacgat gcctgcagca atggcaacaa cgttgcgcaa actattaact ggcgaactac
     7201 ttactctagc ttcccggcaa caattaatag actggatgga ggcggataaa gttgcaggac
     7261 cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct ggagccggtg
     7321 agcgtgggtc tcgcggtatc attgcagcac tggggccaga tggtaagccc tcccgtatcg
     7381 tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga cagatcgctg
     7441 agataggtgc ctcactgatt aagcattggt aactgtcaga ccaagtttac tcatatatac
     7501 tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg
     7561 ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg
     7621 tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc
     7681 aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc
     7741 tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt
     7801 agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc
     7861 taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact
     7921 caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac
     7981 agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag
     8041 aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg
     8101 gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg
     8161 tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga
     8221 gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt
     8281 ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt attaccgcct
     8341 ttgagtgagc tgataccgct cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg
     8401 aggaagcgga agagcgccca atacgcaaac cgcctctccc cgcgcgttgg ccgattcatt
     8461 aatgcagctg gcacgacagg tttcccgact ggaaagcggg cagtgagcgc aacgcaatta
     8521 atgtgagtta gctcactcat taggcacccc aggctttaca ctttatgctt ccggctcgta
     8581 tgttgtgtgg aattgtgagc ggataacaat ttcacacagg aaacagctat gaccatgatt
     8641 acgccaagct ctagctagag gtcgaccaat tctcatgttt gacagcttat catcgcagat
     8701 ccgggcaacg ttgttgccat tgctgcaggc gcagaactgg taggtatgga agatctatac
     8761 attgaatcaa tattggcaat tagccatatt agtcattggt tatatagcat aaatcaatat
     8821 tggctattgg ccattgcata cgttgtatct atatcataat atgtacattt atattggctc
     8881 atgtccaata tgaccgccat gttgacattg attattgact agttattaat agtaatcaat
     8941 tacggggtca ttagttcata gcccatatat ggagttccgc gttacataac ttacggtaaa
     9001 tggcccgcct ggctgaccgc ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt
     9061 tcccatagta acgccaatag ggactttcca ttgacgtcaa tgggtggagt atttacggta
     9121 aactgcccac ttggcagtac atcaagtgta tcatatgcca agtccgcccc ctattgacgt
     9181 caatgacggt aaatggcccg cctggcatta tgcccagtac atgaccttac gggactttcc
     9241 tacttggcag tacatctacg tattagtcat cgctattacc atggtgatgc ggttttggca
     9301 gtacaccaat gggcgtggat agcggtttga ctcacgggga tttccaagtc tccaccccat
     9361 tgacgtcaat gggagtttgt tttggcacca aaatcaacgg gactttccaa aatgtcgtaa
     9421 taaccccgcc ccgttgacgc aaatgggcgg taggcgtgta cggtgggagg tctatataag
     9481 cagagctcgt ttagtgaacc gtcagatcct cactctcttc cgcatcgctg tctgcgaggg
     9541 ccagctgttg ggctcgcggt tgaggacaaa ctcttcgcgg tctttccagt actcttggat
     9601 cggaaacccg tcggcctccg aacggtactc cgccaccgag ggacctgagc gagtccgcat
     9661 cgaccggatc ggaaaacct
//
