LOCUS       sequence-208720-        9142 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 9142)
  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 9142)
  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_0505700-COMP-blac-flag-his
FEATURES             Location/Qualifiers
     source          1..9142
                     /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             2674..3459
                     /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             3466..3531
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             3538..3555
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    3593..3648
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(3647..3666)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(5884..5902)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5970..6074
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6075..6143,6144..6935)
                     /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      7106..7694
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7595..7614
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7848..7865
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    7982..8003
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(8018..8023,8024..8041,8042..8048)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8056..8072
                     /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     8061..8083
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     8080..8096
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     8080..8096
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     enhancer        8367..8746
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        8747..8950
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     8901..8921
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     8947..8971
                     /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 ggccgccgaa
      421 gagtacaaga tcaacaaaga ggaagagaac ggcaagcaca acaaagaaga gatcaagaac
      481 gagaagtacg acaacaacaa ctacgagaac aagaacaatg agaataacaa gaatatcgac
      541 agcaacaacc gcgaggacca catcatgaag ctgctgccca tctacaagga caaggataac
      601 gacatcatcg acgagaacat caacaacaag attaactact atgagaagga caacatcaag
      661 caggaaatca agcacaatac cgagaacgat aacaataata acaacaacat cttcatccag
      721 gaaaaggaca atatccagct gtacgtggac aagaaagagc acctggacga gcggaaccag
      781 aacgtgatcc acgagaatat gaagcaggaa caggaccgga acaaagagac acagaagggc
      841 aacgatatcg acgagtccaa gaagcagaaa gagaaagaac agcacaccaa ccagctgatc
      901 gacaaccgga acgaccagat cagcaacgtg gaaaaagagg acatccacaa cggctacgag
      961 aatgacaagg acaccgagca gaaggatgac aacaaggaag ataagaagga tatcgagcag
     1021 gtcgacgaca ataaggagga caaaaaggac attgaacagg tcgacgataa caaagaggat
     1081 aagaaagaca tcgaacaggt ggacgacaac aaagaagaca aaaaagatat tgagcaggtc
     1141 gatgacaaca aagaagataa gaaagacaca gagcaggtcg acgataacaa agaggacaaa
     1201 aaggatacag aacaggtgga cgataataag gaagataaga agtacattga gcagattgat
     1261 aagaacaagg aggacaagaa ggacaccgag caggtcgacg acaacaaaga ggataagaag
     1321 gaccctcacc agaccgacga caagaacgac ggcgccagct gcgaggaaga aaagaaagtc
     1381 atcatgagcg agattaagac catcaaggag gatctggaca actgcaatat cgccaacaag
     1441 aaagacaaga atgagtgcat cgtgaaagtg aacgaggccg agcggaagct gaacctgtgc
     1501 aaagaaaagg ccctgaagtg caagggccag atcaagggca tccaccacaa gtgcgacgag
     1561 agaatcgacc ggaagaacaa agaaatgatc atcctgcagg gcatcatcaa caagctggaa
     1621 aacaagatcg acgaggataa aaaaaatgat aacaaaaact tccaggatca catcaagaag
     1681 tccgagggca agaagaacga aatccccgtc gaaaacaaaa acaccaagga caacgtgaag
     1741 cgcaacgaga agaatgtgca ccacgaggac tacagcagcc accacaacat ctaccggcag
     1801 gaggataact acctgatcag ctacgagatc ctgaagaact acttcgagaa gatctttgag
     1861 atctacaaga ccctgtacat catcatcgtg gaaaagaccg gcgcgccgtc gacctccatc
     1921 acggcctata agagtgaggg ggagtcagcg gagttctcct tcccactcaa ccttggagag
     1981 gaaagcctgc agggagagtt gagatggaag gcagagaagg ctccttcttc ccagtcctgg
     2041 atcaccttct ccctaaagaa ccaaaaggtg tctgtgcaga agtctactag caaccccaag
     2101 ttccagctgt ccgaaacgct cccactcacc cttcagatac cccaggtctc ccttcagttt
     2161 gctggttctg gcaacctgac cctgactctg gacagaggga tactgtatca ggaagtgaac
     2221 ctggtggtga tgaaagtgac tcagcccgac agcaacactt tgacctgtga ggtgatggga
     2281 cccacctcac ccaagatgag actgatcttg aagcaggaga atcaggaggc cagggtctcc
     2341 aggcaggaga aagtgattca agtgcaggcc cctgaagcag gggtgtggca atgtctactg
     2401 agtgaaggtg aagaggtcaa gatggactcc aagatccagg ttttatccaa agggttgaat
     2461 tccggcggcg gctccggagg cggtaccggt ggcggcgacc tagccccaca gatgcttcga
     2521 gaactccagg agactaatgc ggcgctgcaa gacgtgagag agctcttgcg acagcaggtc
     2581 aaggagatca ccttcctgaa gaatacggtg atggaatgtg acgcctgcgg tctagacaga
     2641 aacctgcctc cactagcacc tctgggccca cacccagaaa cgctggtgaa agtaaaagat
     2701 gctgaagatc agttgggtgc acgagtgggt tacatcgaac tggatctcaa cagcggtaag
     2761 atccttgaga gttttcgccc cgaagaacgt tttccaatga tgagcacttt taaagttctg
     2821 ctatgtggcg cggtattatc ccgtattgac gccgggcaag agcaactcgg tcgccgcata
     2881 cactattctc agaatgactt ggttgagtac tcaccagtca cagaaaagca tcttacggat
     2941 ggcatgacag taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc
     3001 aacttacttc tgacaacgat cggaggaccg aaggagctaa ccgctttttt gcacaacatg
     3061 ggggatcatg taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac
     3121 gacgagcgtg acaccacgat gcctgtagca atggcaacaa cgttgcgcaa actattaact
     3181 ggcgaactac ttactctagc ttcccggcaa caattaatag actggatgga ggcggataaa
     3241 gttgcaggac cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct
     3301 ggagccggtg agcgtgggtc tcgcggtatc attgcagcac tggggccaga tggtaagccc
     3361 tcccgtatcg tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga
     3421 cagatcgctg agataggtgc ctcactgatt aagcattggg gccctgacta caaagaccat
     3481 gacggtgatt ataaagatca tgacatcgat tacaaggatg acgatgacaa gggccctcac
     3541 caccaccatc accactaatt aattaaggat cccccgacct cgacctctgg ctaataaagg
     3601 aaatttattt tcattgcaat agtgtgttgg aattttttgt gtctctcact cggaaggaca
     3661 tatgggaggg caaatcattt ggtcgagatc cctcggagat ctctagctag aggatcgatc
     3721 cccgccccgg acgaactaaa cctgactacg acatctctgc cccttcttcg cggggcagtg
     3781 catgtaatcc cttcagttgg ttggtacaac ttgccaactg ggccctgttc cacatgtgac
     3841 acgggggggg accaaacaca aaggggttct ctgactgtag ttgacatcct tataaatgga
     3901 tgtgcacatt tgccaacact gagtggcttt catcctggag cagactttgc agtctgtgga
     3961 ctgcaacaca acattgcctt tatgtgtaac tcttggctga agctcttaca ccaatgctgg
     4021 gggacatgta cctcccaggg gcccaggaag actacgggag gctacaccaa cgtcaatcag
     4081 aggggcctgt gtagctaccg ataagcggac cctcaagagg gcattagcaa tagtgtttat
     4141 aaggccccct tgttaaccct aaacgggtag catatgcttc ccgggtagta gtatatacta
     4201 tccagactaa ccctaattca atagcatatg ttacccaacg ggaagcatat gctatcgaat
     4261 tagggttagt aaaagggtcc taaggaacag cgatatctcc caccccatga gctgtcacgg
     4321 ttttatttac atggggtcag gattccacga gggtagtgaa ccattttagt cacaagggca
     4381 gtggctgaag atcaaggagc gggcagtgaa ctctcctgaa tcttcgcctg cttcttcatt
     4441 ctccttcgtt tagctaatag aataactgct gagttgtgaa cagtaaggtg tatgtgaggt
     4501 gctcgaaaac aaggtttcag gtgacgcccc cagaataaaa tttggacggg gggttcagtg
     4561 gtggcattgt gctatgacac caatataacc ctcacaaacc ccttgggcaa taaatactag
     4621 tgtaggaatg aaacattctg aatatcttta acaatagaaa tccatggggt ggggacaagc
     4681 cgtaaagact ggatgtccat ctcacacgaa tttatggcta tgggcaacac ataatcctag
     4741 tgcaatatga tactggggtt attaagatgt gtcccaggca gggaccaaga caggtgaacc
     4801 atgttgttac actctatttg taacaagggg aaagagagtg gacgccgaca gcagcggact
     4861 ccactggttg tctctaacac ccccgaaaat taaacggggc tccacgccaa tggggcccat
     4921 aaacaaagac aagtggccac tctttttttt gaaattgtgg agtgggggca cgcgtcagcc
     4981 cccacacgcc gccctgcggt tttggactgt aaaataaggg tgtaataact tggctgattg
     5041 taaccccgct aaccactgcg gtcaaaccac ttgcccacaa aaccactaat ggcaccccgg
     5101 ggaatacctg cataagtagg tgggcgggcc aagatagggg cgcgattgct gcgatctgga
     5161 ggacaaatta cacacacttg cgcctgagcg ccaagcacag ggttgttggt cctcatattc
     5221 acgaggtcgc tgagagcacg gtgggctaat gttgccatgg gtagcatata ctacccaaat
     5281 atctggatag catatgctat cctaatctat atctgggtag cataggctat cctaatctat
     5341 atctgggtag catatgctat cctaatctat atctgggtag tatatgctat cctaatttat
     5401 atctgggtag cataggctat cctaatctat atctgggtag catatgctat cctaatctat
     5461 atctgggtag tatatgctat cctaatctgt atccgggtag catatgctat cctaatagag
     5521 attagggtag tatatgctat cctaatttat atctgggtag catatactac ccaaatatct
     5581 ggatagcata tgctatccta atctatatct gggtagcata tgctatccta atctatatct
     5641 gggtagcata ggctatccta atctatatct gggtagcata tgctatccta atctatatct
     5701 gggtagtata tgctatccta atttatatct gggtagcata ggctatccta atctatatct
     5761 gggtagcata tgctatccta atctatatct gggtagtata tgctatccta atctgtatcc
     5821 gggtagcata tgctatcctc atgataagct gtcaaacatg agaattttct tgaagacgaa
     5881 agggcctcgt gatacgccta tttttatagg ttaatgtcat gataataatg gtttcttaga
     5941 cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa
     6001 tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt
     6061 gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg
     6121 cattttgcct tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctgaag
     6181 atcagttggg tgcacgagtg ggttacatcg aactggatct caacagcggt aagatccttg
     6241 agagttttcg ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg
     6301 gcgcggtatt atcccgtgtt gacgccgggc aagagcaact cggtcgccgc atacactatt
     6361 ctcagaatga cttggttgag tactcaccag tcacagaaaa gcatcttacg gatggcatga
     6421 cagtaagaga attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac
     6481 ttctgacaac gatcggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc
     6541 atgtaactcg ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc
     6601 gtgacaccac gatgcctgca gcaatggcaa caacgttgcg caaactatta actggcgaac
     6661 tacttactct agcttcccgg caacaattaa tagactggat ggaggcggat aaagttgcag
     6721 gaccacttct gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg
     6781 gtgagcgtgg gtctcgcggt atcattgcag cactggggcc agatggtaag ccctcccgta
     6841 tcgtagttat ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg
     6901 ctgagatagg tgcctcactg attaagcatt ggtaactgtc agaccaagtt tactcatata
     6961 tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt
     7021 ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc
     7081 ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct
     7141 tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa
     7201 ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag
     7261 tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc
     7321 tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg
     7381 actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca
     7441 cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat
     7501 gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg
     7561 tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc
     7621 ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc
     7681 ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc
     7741 cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg
     7801 cctttgagtg agctgatacc gctcgccgca gccgaacgac cgagcgcagc gagtcagtga
     7861 gcgaggaagc ggaagagcgc ccaatacgca aaccgcctct ccccgcgcgt tggccgattc
     7921 attaatgcag ctggcacgac aggtttcccg actggaaagc gggcagtgag cgcaacgcaa
     7981 ttaatgtgag ttagctcact cattaggcac cccaggcttt acactttatg cttccggctc
     8041 gtatgttgtg tggaattgtg agcggataac aatttcacac aggaaacagc tatgaccatg
     8101 attacgccaa gctctagcta gaggtcgacc aattctcatg tttgacagct tatcatcgca
     8161 gatccgggca acgttgttgc cattgctgca ggcgcagaac tggtaggtat ggaagatcta
     8221 tacattgaat caatattggc aattagccat attagtcatt ggttatatag cataaatcaa
     8281 tattggctat tggccattgc atacgttgta tctatatcat aatatgtaca tttatattgg
     8341 ctcatgtcca atatgaccgc catgttgaca ttgattattg actagttatt aatagtaatc
     8401 aattacgggg tcattagttc atagcccata tatggagttc cgcgttacat aacttacggt
     8461 aaatggcccg cctggctgac cgcccaacga cccccgccca ttgacgtcaa taatgacgta
     8521 tgttcccata gtaacgccaa tagggacttt ccattgacgt caatgggtgg agtatttacg
     8581 gtaaactgcc cacttggcag tacatcaagt gtatcatatg ccaagtccgc cccctattga
     8641 cgtcaatgac ggtaaatggc ccgcctggca ttatgcccag tacatgacct tacgggactt
     8701 tcctacttgg cagtacatct acgtattagt catcgctatt accatggtga tgcggttttg
     8761 gcagtacacc aatgggcgtg gatagcggtt tgactcacgg ggatttccaa gtctccaccc
     8821 cattgacgtc aatgggagtt tgttttggca ccaaaatcaa cgggactttc caaaatgtcg
     8881 taataacccc gccccgttga cgcaaatggg cggtaggcgt gtacggtggg aggtctatat
     8941 aagcagagct cgtttagtga accgtcagat cctcactctc ttccgcatcg ctgtctgcga
     9001 gggccagctg ttgggctcgc ggttgaggac aaactcttcg cggtctttcc agtactcttg
     9061 gatcggaaac ccgtcggcct ccgaacggta ctccgccacc gagggacctg agcgagtccg
     9121 catcgaccgg atcggaaaac ct
//
