LOCUS       sequence-224479-        8459 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 8459)
  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 8459)
  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: WARP-COMP-blac-flag-his
FEATURES             Location/Qualifiers
     source          1..8459
                     /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             1993..2778
                     /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             2785..2850
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             2857..2874
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    2910..2965
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(2964..2983)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(5201..5219)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5287..5391
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5392..5460,5461..6252)
                     /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      6423..7011
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6912..6931
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7165..7182
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    7299..7320
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7335..7340,7341..7358,7359..7365)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7373..7389
                     /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     7378..7400
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7397..7413
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7397..7413
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     enhancer        7684..8063
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        8064..8267
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     8218..8238
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     8264..8288
                     /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 ggccgccaga
      421 atgaacgtgg tgtcccacaa cagcatgaag aaagtgggcg aggaaggcaa ggacgagaag
      481 atcatcagct gcgtggtgga atacgtgatc aagggcgacc tgagcatcga cgacggcgat
      541 ttctgcgaca gcaacagcat ctgcagcaac aaccccggcc tgtgcgacaa cggccaggtg
      601 gtggtgcccg gcgactactg ctccaactac tacgacatca ccctggtggt ggaagagtcc
      661 agcttcgtgc agaaggacta ctggctgaag ggcaccatcc cctttctgga atccatggtg
      721 ttcaacagca acgtggccaa ggacaaggcc cacatgagcg tggtgctgtt cgccggcaag
      781 cagcgggtgg tggtcccctt caccgacgag agcagccagg acaagaacat gctgctggaa
      841 aagatcaact ccatcgacga gggaatcgcc accagcagcg acaccctgta cgtgaacgcc
      901 ctgaagtacg ccatcgagaa cgtgatcttc ggcgacggca cccggaagga cgcccctaag
      961 gtggccgtgc tgttctacta cggcttcgac tacggcgcca acaagaatat gatccccgac
     1021 gtggtgcaga cctacaaaga acagaacgtg aagctgatca tcgtggggat cgccctgggc
     1081 atccgggaca acgcctacct gctggccgac tgcagcatcg gcgacgagaa ctgtgtgaat
     1141 gtggtgttta agccctggga cttcgtgatc cccgctgccg ccgacgtgaa ccagaagatc
     1201 tgcgataaga acaaggccgg cgcgccgtcg acctccatca cggcctataa gagtgagggg
     1261 gagtcagcgg agttctcctt cccactcaac cttggagagg aaagcctgca gggagagttg
     1321 agatggaagg cagagaaggc tccttcttcc cagtcctgga tcaccttctc cctaaagaac
     1381 caaaaggtgt ctgtgcagaa gtctactagc aaccccaagt tccagctgtc cgaaacgctc
     1441 ccactcaccc ttcagatacc ccaggtctcc cttcagtttg ctggttctgg caacctgacc
     1501 ctgactctgg acagagggat actgtatcag gaagtgaacc tggtggtgat gaaagtgact
     1561 cagcccgaca gcaacacttt gacctgtgag gtgatgggac ccacctcacc caagatgaga
     1621 ctgatcttga agcaggagaa tcaggaggcc agggtctcca ggcaggagaa agtgattcaa
     1681 gtgcaggccc ctgaagcagg ggtgtggcaa tgtctactga gtgaaggtga agaggtcaag
     1741 atggactcca agatccaggt tttatccaaa gggttgaatt ccggcggcgg ctccggaggc
     1801 ggtaccggtg gcggcgacct agccccacag atgcttcgag aactccagga gactaatgcg
     1861 gcgctgcaag acgtgagaga gctcttgcga cagcaggtca aggagatcac cttcctgaag
     1921 aatacggtga tggaatgtga cgcctgcggt ctagacagaa acctgcctcc actagcacct
     1981 ctgggcccac acccagaaac gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca
     2041 cgagtgggtt acatcgaact ggatctcaac agcggtaaga tccttgagag ttttcgcccc
     2101 gaagaacgtt ttccaatgat gagcactttt aaagttctgc tatgtggcgc ggtattatcc
     2161 cgtattgacg ccgggcaaga gcaactcggt cgccgcatac actattctca gaatgacttg
     2221 gttgagtact caccagtcac agaaaagcat cttacggatg gcatgacagt aagagaatta
     2281 tgcagtgctg ccataaccat gagtgataac actgcggcca acttacttct gacaacgatc
     2341 ggaggaccga aggagctaac cgcttttttg cacaacatgg gggatcatgt aactcgcctt
     2401 gatcgttggg aaccggagct gaatgaagcc ataccaaacg acgagcgtga caccacgatg
     2461 cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg gcgaactact tactctagct
     2521 tcccggcaac aattaataga ctggatggag gcggataaag ttgcaggacc acttctgcgc
     2581 tcggcccttc cggctggctg gtttattgct gataaatctg gagccggtga gcgtgggtct
     2641 cgcggtatca ttgcagcact ggggccagat ggtaagccct cccgtatcgt agttatctac
     2701 acgacgggga gtcaggcaac tatggatgaa cgaaatagac agatcgctga gataggtgcc
     2761 tcactgatta agcattgggg ccctgactac aaagaccatg acggtgatta taaagatcat
     2821 gacatcgatt acaaggatga cgatgacaag ggccctcacc accaccatca ccactaataa
     2881 taaggatccc ccgacctcga cctctggcta ataaaggaaa tttattttca ttgcaatagt
     2941 gtgttggaat tttttgtgtc tctcactcgg aaggacatat gggagggcaa atcatttggt
     3001 cgagatccct cggagatctc tagctagagg atcgatcccc gccccggacg aactaaacct
     3061 gactacgaca tctctgcccc ttcttcgcgg ggcagtgcat gtaatccctt cagttggttg
     3121 gtacaacttg ccaactgggc cctgttccac atgtgacacg gggggggacc aaacacaaag
     3181 gggttctctg actgtagttg acatccttat aaatggatgt gcacatttgc caacactgag
     3241 tggctttcat cctggagcag actttgcagt ctgtggactg caacacaaca ttgcctttat
     3301 gtgtaactct tggctgaagc tcttacacca atgctggggg acatgtacct cccaggggcc
     3361 caggaagact acgggaggct acaccaacgt caatcagagg ggcctgtgta gctaccgata
     3421 agcggaccct caagagggca ttagcaatag tgtttataag gcccccttgt taaccctaaa
     3481 cgggtagcat atgcttcccg ggtagtagta tatactatcc agactaaccc taattcaata
     3541 gcatatgtta cccaacggga agcatatgct atcgaattag ggttagtaaa agggtcctaa
     3601 ggaacagcga tatctcccac cccatgagct gtcacggttt tatttacatg gggtcaggat
     3661 tccacgaggg tagtgaacca ttttagtcac aagggcagtg gctgaagatc aaggagcggg
     3721 cagtgaactc tcctgaatct tcgcctgctt cttcattctc cttcgtttag ctaatagaat
     3781 aactgctgag ttgtgaacag taaggtgtat gtgaggtgct cgaaaacaag gtttcaggtg
     3841 acgcccccag aataaaattt ggacgggggg ttcagtggtg gcattgtgct atgacaccaa
     3901 tataaccctc acaaacccct tgggcaataa atactagtgt aggaatgaaa cattctgaat
     3961 atctttaaca atagaaatcc atggggtggg gacaagccgt aaagactgga tgtccatctc
     4021 acacgaattt atggctatgg gcaacacata atcctagtgc aatatgatac tggggttatt
     4081 aagatgtgtc ccaggcaggg accaagacag gtgaaccatg ttgttacact ctatttgtaa
     4141 caaggggaaa gagagtggac gccgacagca gcggactcca ctggttgtct ctaacacccc
     4201 cgaaaattaa acggggctcc acgccaatgg ggcccataaa caaagacaag tggccactct
     4261 tttttttgaa attgtggagt gggggcacgc gtcagccccc acacgccgcc ctgcggtttt
     4321 ggactgtaaa ataagggtgt aataacttgg ctgattgtaa ccccgctaac cactgcggtc
     4381 aaaccacttg cccacaaaac cactaatggc accccgggga atacctgcat aagtaggtgg
     4441 gcgggccaag ataggggcgc gattgctgcg atctggagga caaattacac acacttgcgc
     4501 ctgagcgcca agcacagggt tgttggtcct catattcacg aggtcgctga gagcacggtg
     4561 ggctaatgtt gccatgggta gcatatacta cccaaatatc tggatagcat atgctatcct
     4621 aatctatatc tgggtagcat aggctatcct aatctatatc tgggtagcat atgctatcct
     4681 aatctatatc tgggtagtat atgctatcct aatttatatc tgggtagcat aggctatcct
     4741 aatctatatc tgggtagcat atgctatcct aatctatatc tgggtagtat atgctatcct
     4801 aatctgtatc cgggtagcat atgctatcct aatagagatt agggtagtat atgctatcct
     4861 aatttatatc tgggtagcat atactaccca aatatctgga tagcatatgc tatcctaatc
     4921 tatatctggg tagcatatgc tatcctaatc tatatctggg tagcataggc tatcctaatc
     4981 tatatctggg tagcatatgc tatcctaatc tatatctggg tagtatatgc tatcctaatt
     5041 tatatctggg tagcataggc tatcctaatc tatatctggg tagcatatgc tatcctaatc
     5101 tatatctggg tagtatatgc tatcctaatc tgtatccggg tagcatatgc tatcctcatg
     5161 ataagctgtc aaacatgaga attttcttga agacgaaagg gcctcgtgat acgcctattt
     5221 ttataggtta atgtcatgat aataatggtt tcttagacgt caggtggcac ttttcgggga
     5281 aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc
     5341 atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag tatgagtatt
     5401 caacatttcc gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct
     5461 cacccagaaa cgctggtgaa agtaaaagat gctgaagatc agttgggtgc acgagtgggt
     5521 tacatcgaac tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgt
     5581 tttccaatga tgagcacttt taaagttctg ctatgtggcg cggtattatc ccgtgttgac
     5641 gccgggcaag agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtac
     5701 tcaccagtca cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct
     5761 gccataacca tgagtgataa cactgcggcc aacttacttc tgacaacgat cggaggaccg
     5821 aaggagctaa ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg
     5881 gaaccggagc tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgcagca
     5941 atggcaacaa cgttgcgcaa actattaact ggcgaactac ttactctagc ttcccggcaa
     6001 caattaatag actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt
     6061 ccggctggct ggtttattgc tgataaatct ggagccggtg agcgtgggtc tcgcggtatc
     6121 attgcagcac tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg
     6181 agtcaggcaa ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt
     6241 aagcattggt aactgtcaga ccaagtttac tcatatatac tttagattga tttaaaactt
     6301 catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc
     6361 ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct
     6421 tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta
     6481 ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc
     6541 ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt aggccaccac
     6601 ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct
     6661 gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat
     6721 aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg
     6781 acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa
     6841 gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg
     6901 gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga
     6961 cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc
     7021 aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct
     7081 gcgttatccc ctgattctgt ggataaccgt attaccgcct ttgagtgagc tgataccgct
     7141 cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga agagcgccca
     7201 atacgcaaac cgcctctccc cgcgcgttgg ccgattcatt aatgcagctg gcacgacagg
     7261 tttcccgact ggaaagcggg cagtgagcgc aacgcaatta atgtgagtta gctcactcat
     7321 taggcacccc aggctttaca ctttatgctt ccggctcgta tgttgtgtgg aattgtgagc
     7381 ggataacaat ttcacacagg aaacagctat gaccatgatt acgccaagct ctagctagag
     7441 gtcgaccaat tctcatgttt gacagcttat catcgcagat ccgggcaacg ttgttgccat
     7501 tgctgcaggc gcagaactgg taggtatgga agatctatac attgaatcaa tattggcaat
     7561 tagccatatt agtcattggt tatatagcat aaatcaatat tggctattgg ccattgcata
     7621 cgttgtatct atatcataat atgtacattt atattggctc atgtccaata tgaccgccat
     7681 gttgacattg attattgact agttattaat agtaatcaat tacggggtca ttagttcata
     7741 gcccatatat ggagttccgc gttacataac ttacggtaaa tggcccgcct ggctgaccgc
     7801 ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt tcccatagta acgccaatag
     7861 ggactttcca ttgacgtcaa tgggtggagt atttacggta aactgcccac ttggcagtac
     7921 atcaagtgta tcatatgcca agtccgcccc ctattgacgt caatgacggt aaatggcccg
     7981 cctggcatta tgcccagtac atgaccttac gggactttcc tacttggcag tacatctacg
     8041 tattagtcat cgctattacc atggtgatgc ggttttggca gtacaccaat gggcgtggat
     8101 agcggtttga ctcacgggga tttccaagtc tccaccccat tgacgtcaat gggagtttgt
     8161 tttggcacca aaatcaacgg gactttccaa aatgtcgtaa taaccccgcc ccgttgacgc
     8221 aaatgggcgg taggcgtgta cggtgggagg tctatataag cagagctcgt ttagtgaacc
     8281 gtcagatcct cactctcttc cgcatcgctg tctgcgaggg ccagctgttg ggctcgcggt
     8341 tgaggacaaa ctcttcgcgg tctttccagt actcttggat cggaaacccg tcggcctccg
     8401 aacggtactc cgccaccgag ggacctgagc gagtccgcat cgaccggatc ggaaaacct
//
