LOCUS       sequence-208818-        8482 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 8482)
  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 8482)
  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: PF3D7_1404900-COMP-blac-flag-his
FEATURES             Location/Qualifiers
     source          1..8482
                     /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             2014..2799
                     /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             2806..2871
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             2878..2895
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    2933..2988
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(2987..3006)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(5224..5242)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5310..5414
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5415..5483,5484..6275)
                     /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      6446..7034
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6935..6954
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7188..7205
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    7322..7343
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7358..7363,7364..7381,7382..7388)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7396..7412
                     /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     7401..7423
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7420..7436
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7420..7436
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     enhancer        7707..8086
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        8087..8290
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     8241..8261
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     8287..8311
                     /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 accgagagca agaacatcct gaacaccgtg aagaacgacg tcaagaacgg caactaccag
      481 aacatgatga acttccccga catggacaag atcaaagagc tgatcaacca gaacaaggtg
      541 gaagaggccc gcgagctgtt caacgagatg cagaaacagt tcatgaagga aaagatggaa
      601 gccaacaaca acatgaacaa caagaagccc gccaagaaag ccatggacaa cagcaacctg
      661 ttcagcaacc tcatcaaccc caacgacctg caggaaatga tcaagttcta catgtacctg
      721 cagaatatcc tcaacagcaa gaacgaggcc agcgagaaga agatggtcaa gagcttcctg
      781 cgcaagacca tccagaagat gaaggacaac gagaacagcg aggaaaagaa gtccatcgac
      841 gacatcatca aggacaccga gggccacaat gagatgctgg aaaagctggc cggcctgatg
      901 aagatcaaca aagagaagct gaaagacgag caggtcaaaa acaagctgaa ccagatcctg
      961 atcggcatgc tcaagttcat cgactacacc aacaacgccg acatcaccaa cagcctgatg
     1021 gacgagatcc agatcgaaca ggtgcaggcc gaggacggca ccatcaagcc caaggtgcag
     1081 gtcaacattg ccaactccaa ggcccacctg gacatgctgc agaaagccac caacttcatg
     1141 ggcatcaaca tcgaccagga agaactgaag aagctgaccg tcaacaacaa gtggtacgag
     1201 tccttcctga acaacctgct gaactccgcc gacgaactcg gcgcgccgtc gacctccatc
     1261 acggcctata agagtgaggg ggagtcagcg gagttctcct tcccactcaa ccttggagag
     1321 gaaagcctgc agggagagtt gagatggaag gcagagaagg ctccttcttc ccagtcctgg
     1381 atcaccttct ccctaaagaa ccaaaaggtg tctgtgcaga agtctactag caaccccaag
     1441 ttccagctgt ccgaaacgct cccactcacc cttcagatac cccaggtctc ccttcagttt
     1501 gctggttctg gcaacctgac cctgactctg gacagaggga tactgtatca ggaagtgaac
     1561 ctggtggtga tgaaagtgac tcagcccgac agcaacactt tgacctgtga ggtgatggga
     1621 cccacctcac ccaagatgag actgatcttg aagcaggaga atcaggaggc cagggtctcc
     1681 aggcaggaga aagtgattca agtgcaggcc cctgaagcag gggtgtggca atgtctactg
     1741 agtgaaggtg aagaggtcaa gatggactcc aagatccagg ttttatccaa agggttgaat
     1801 tccggcggcg gctccggagg cggtaccggt ggcggcgacc tagccccaca gatgcttcga
     1861 gaactccagg agactaatgc ggcgctgcaa gacgtgagag agctcttgcg acagcaggtc
     1921 aaggagatca ccttcctgaa gaatacggtg atggaatgtg acgcctgcgg tctagacaga
     1981 aacctgcctc cactagcacc tctgggccca cacccagaaa cgctggtgaa agtaaaagat
     2041 gctgaagatc agttgggtgc acgagtgggt tacatcgaac tggatctcaa cagcggtaag
     2101 atccttgaga gttttcgccc cgaagaacgt tttccaatga tgagcacttt taaagttctg
     2161 ctatgtggcg cggtattatc ccgtattgac gccgggcaag agcaactcgg tcgccgcata
     2221 cactattctc agaatgactt ggttgagtac tcaccagtca cagaaaagca tcttacggat
     2281 ggcatgacag taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc
     2341 aacttacttc tgacaacgat cggaggaccg aaggagctaa ccgctttttt gcacaacatg
     2401 ggggatcatg taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac
     2461 gacgagcgtg acaccacgat gcctgtagca atggcaacaa cgttgcgcaa actattaact
     2521 ggcgaactac ttactctagc ttcccggcaa caattaatag actggatgga ggcggataaa
     2581 gttgcaggac cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct
     2641 ggagccggtg agcgtgggtc tcgcggtatc attgcagcac tggggccaga tggtaagccc
     2701 tcccgtatcg tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga
     2761 cagatcgctg agataggtgc ctcactgatt aagcattggg gccctgacta caaagaccat
     2821 gacggtgatt ataaagatca tgacatcgat tacaaggatg acgatgacaa gggccctcac
     2881 caccaccatc accactaatt aattaaggat cccccgacct cgacctctgg ctaataaagg
     2941 aaatttattt tcattgcaat agtgtgttgg aattttttgt gtctctcact cggaaggaca
     3001 tatgggaggg caaatcattt ggtcgagatc cctcggagat ctctagctag aggatcgatc
     3061 cccgccccgg acgaactaaa cctgactacg acatctctgc cccttcttcg cggggcagtg
     3121 catgtaatcc cttcagttgg ttggtacaac ttgccaactg ggccctgttc cacatgtgac
     3181 acgggggggg accaaacaca aaggggttct ctgactgtag ttgacatcct tataaatgga
     3241 tgtgcacatt tgccaacact gagtggcttt catcctggag cagactttgc agtctgtgga
     3301 ctgcaacaca acattgcctt tatgtgtaac tcttggctga agctcttaca ccaatgctgg
     3361 gggacatgta cctcccaggg gcccaggaag actacgggag gctacaccaa cgtcaatcag
     3421 aggggcctgt gtagctaccg ataagcggac cctcaagagg gcattagcaa tagtgtttat
     3481 aaggccccct tgttaaccct aaacgggtag catatgcttc ccgggtagta gtatatacta
     3541 tccagactaa ccctaattca atagcatatg ttacccaacg ggaagcatat gctatcgaat
     3601 tagggttagt aaaagggtcc taaggaacag cgatatctcc caccccatga gctgtcacgg
     3661 ttttatttac atggggtcag gattccacga gggtagtgaa ccattttagt cacaagggca
     3721 gtggctgaag atcaaggagc gggcagtgaa ctctcctgaa tcttcgcctg cttcttcatt
     3781 ctccttcgtt tagctaatag aataactgct gagttgtgaa cagtaaggtg tatgtgaggt
     3841 gctcgaaaac aaggtttcag gtgacgcccc cagaataaaa tttggacggg gggttcagtg
     3901 gtggcattgt gctatgacac caatataacc ctcacaaacc ccttgggcaa taaatactag
     3961 tgtaggaatg aaacattctg aatatcttta acaatagaaa tccatggggt ggggacaagc
     4021 cgtaaagact ggatgtccat ctcacacgaa tttatggcta tgggcaacac ataatcctag
     4081 tgcaatatga tactggggtt attaagatgt gtcccaggca gggaccaaga caggtgaacc
     4141 atgttgttac actctatttg taacaagggg aaagagagtg gacgccgaca gcagcggact
     4201 ccactggttg tctctaacac ccccgaaaat taaacggggc tccacgccaa tggggcccat
     4261 aaacaaagac aagtggccac tctttttttt gaaattgtgg agtgggggca cgcgtcagcc
     4321 cccacacgcc gccctgcggt tttggactgt aaaataaggg tgtaataact tggctgattg
     4381 taaccccgct aaccactgcg gtcaaaccac ttgcccacaa aaccactaat ggcaccccgg
     4441 ggaatacctg cataagtagg tgggcgggcc aagatagggg cgcgattgct gcgatctgga
     4501 ggacaaatta cacacacttg cgcctgagcg ccaagcacag ggttgttggt cctcatattc
     4561 acgaggtcgc tgagagcacg gtgggctaat gttgccatgg gtagcatata ctacccaaat
     4621 atctggatag catatgctat cctaatctat atctgggtag cataggctat cctaatctat
     4681 atctgggtag catatgctat cctaatctat atctgggtag tatatgctat cctaatttat
     4741 atctgggtag cataggctat cctaatctat atctgggtag catatgctat cctaatctat
     4801 atctgggtag tatatgctat cctaatctgt atccgggtag catatgctat cctaatagag
     4861 attagggtag tatatgctat cctaatttat atctgggtag catatactac ccaaatatct
     4921 ggatagcata tgctatccta atctatatct gggtagcata tgctatccta atctatatct
     4981 gggtagcata ggctatccta atctatatct gggtagcata tgctatccta atctatatct
     5041 gggtagtata tgctatccta atttatatct gggtagcata ggctatccta atctatatct
     5101 gggtagcata tgctatccta atctatatct gggtagtata tgctatccta atctgtatcc
     5161 gggtagcata tgctatcctc atgataagct gtcaaacatg agaattttct tgaagacgaa
     5221 agggcctcgt gatacgccta tttttatagg ttaatgtcat gataataatg gtttcttaga
     5281 cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa
     5341 tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt
     5401 gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg
     5461 cattttgcct tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctgaag
     5521 atcagttggg tgcacgagtg ggttacatcg aactggatct caacagcggt aagatccttg
     5581 agagttttcg ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg
     5641 gcgcggtatt atcccgtgtt gacgccgggc aagagcaact cggtcgccgc atacactatt
     5701 ctcagaatga cttggttgag tactcaccag tcacagaaaa gcatcttacg gatggcatga
     5761 cagtaagaga attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac
     5821 ttctgacaac gatcggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc
     5881 atgtaactcg ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc
     5941 gtgacaccac gatgcctgca gcaatggcaa caacgttgcg caaactatta actggcgaac
     6001 tacttactct agcttcccgg caacaattaa tagactggat ggaggcggat aaagttgcag
     6061 gaccacttct gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg
     6121 gtgagcgtgg gtctcgcggt atcattgcag cactggggcc agatggtaag ccctcccgta
     6181 tcgtagttat ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg
     6241 ctgagatagg tgcctcactg attaagcatt ggtaactgtc agaccaagtt tactcatata
     6301 tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt
     6361 ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc
     6421 ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct
     6481 tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa
     6541 ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag
     6601 tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc
     6661 tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg
     6721 actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca
     6781 cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat
     6841 gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg
     6901 tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc
     6961 ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc
     7021 ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc
     7081 cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg
     7141 cctttgagtg agctgatacc gctcgccgca gccgaacgac cgagcgcagc gagtcagtga
     7201 gcgaggaagc ggaagagcgc ccaatacgca aaccgcctct ccccgcgcgt tggccgattc
     7261 attaatgcag ctggcacgac aggtttcccg actggaaagc gggcagtgag cgcaacgcaa
     7321 ttaatgtgag ttagctcact cattaggcac cccaggcttt acactttatg cttccggctc
     7381 gtatgttgtg tggaattgtg agcggataac aatttcacac aggaaacagc tatgaccatg
     7441 attacgccaa gctctagcta gaggtcgacc aattctcatg tttgacagct tatcatcgca
     7501 gatccgggca acgttgttgc cattgctgca ggcgcagaac tggtaggtat ggaagatcta
     7561 tacattgaat caatattggc aattagccat attagtcatt ggttatatag cataaatcaa
     7621 tattggctat tggccattgc atacgttgta tctatatcat aatatgtaca tttatattgg
     7681 ctcatgtcca atatgaccgc catgttgaca ttgattattg actagttatt aatagtaatc
     7741 aattacgggg tcattagttc atagcccata tatggagttc cgcgttacat aacttacggt
     7801 aaatggcccg cctggctgac cgcccaacga cccccgccca ttgacgtcaa taatgacgta
     7861 tgttcccata gtaacgccaa tagggacttt ccattgacgt caatgggtgg agtatttacg
     7921 gtaaactgcc cacttggcag tacatcaagt gtatcatatg ccaagtccgc cccctattga
     7981 cgtcaatgac ggtaaatggc ccgcctggca ttatgcccag tacatgacct tacgggactt
     8041 tcctacttgg cagtacatct acgtattagt catcgctatt accatggtga tgcggttttg
     8101 gcagtacacc aatgggcgtg gatagcggtt tgactcacgg ggatttccaa gtctccaccc
     8161 cattgacgtc aatgggagtt tgttttggca ccaaaatcaa cgggactttc caaaatgtcg
     8221 taataacccc gccccgttga cgcaaatggg cggtaggcgt gtacggtggg aggtctatat
     8281 aagcagagct cgtttagtga accgtcagat cctcactctc ttccgcatcg ctgtctgcga
     8341 gggccagctg ttgggctcgc ggttgaggac aaactcttcg cggtctttcc agtactcttg
     8401 gatcggaaac ccgtcggcct ccgaacggta ctccgccacc gagggacctg agcgagtccg
     8461 catcgaccgg atcggaaaac ct
//
