LOCUS       sequence-224705-        9422 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 9422)
  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 9422)
  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_0624400-COMP-blac-flag-his
FEATURES             Location/Qualifiers
     source          1..9422
                     /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             2956..3741
                     /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             3748..3813
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             3820..3837
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    3873..3928
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(3927..3946)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(6164..6182)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6250..6354
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6355..6423,6424..7215)
                     /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      7386..7974
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7875..7894
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     8128..8145
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    8262..8283
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(8298..8303,8304..8321,8322..8328)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8336..8352
                     /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     8341..8363
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     8360..8376
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     8360..8376
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     enhancer        8647..9026
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        9027..9230
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     9181..9201
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     9227..9251
                     /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 gagagaaaga tcggcgagca gtacgagtac atcaacctgc tgagcgacaa agagtaccgg
      481 aacaagctgc tggccctgcg gaagcggacc ttcaacacca agaataccaa gatcaacaac
      541 aataataaca ataacaacat tgccaacagc aacggcaaca aagtgaacga catctacaac
      601 aagaaaaact acgtgttctg cttctacggc gcctgcggct acgacaagcg gaacgtgctg
      661 gaaaagatct tccacatcac cctggaaaag gaccagctga tcaacctggg ctacaaccag
      721 cggctgggca tttggttcgg caaggacaag gacgagaacc tgaacatgat cctggacatc
      781 gacctgctcg aaaacaaaga gttcatcgag aagaacaacg aggccatcta caattacgag
      841 aagctgatcg agacaatcct caagaccacc aacaccatca tcatccctct gctgtacgag
      901 gacatccagt gcgtgaccca cgtgctgaac aagaacaaca agatcaccga cgggaagaac
      961 aaggagtaca acaatctgaa gtacaagctg cctatcaaga tcgagctgtt cctgaacgag
     1021 ctgaaccgga agatcgagga agagaagaaa aagaacgaga acgactactc caacgggaag
     1081 gacatcctcg acaaaaaaaa gaacaatgtg tacttcatct tcctcaacaa cgagaaagag
     1141 aacaagcgca actacgccta caacaacaac ttcatccagg aaatcaagct gcggtacaac
     1201 aacctcaact tctacttcat gaacgaaaaa gagatcaacc tcgccctgct gaagtccaac
     1261 aatatcgccg agtgcatcat cctgatggac aacatcgagc acgccctgaa ggacatgagc
     1321 accttcaaga acttcgccga cttcaacaag gcctgcgtgt acaataccta cgtgatcgaa
     1381 gagagctaca tcaagaccct gaactacttc gacaagtact tctacaccta cgagaagaat
     1441 atcgccatgg gcaagatcat taacaagtac aacatcctga gcaacgaact gatcaagaac
     1501 agcctgctgc tgttcaatct gctgaccctg gaacagagcg gcaccaagtt caagcagatc
     1561 atcttcgaga agctcaagaa tcggttccag atgagcatcc ggcggcagat cctgaagcag
     1621 ctcctgctgc tcgagaaaaa gtatatcgag aagggcaagc acatcattct gaacaagtac
     1681 tatatcaaga atgacaaaga gctgattaac aacgagcaca accagctgca catcctgaag
     1741 aaaaccctga acagccagat caagcaggat atccagaccc tgatccacaa gcagttctac
     1801 aacgagcccc actttaacaa cgccatcaac cagttcatct accagctcga ggacaagctg
     1861 gaccacaccc tcaacaatta ctatgagctg aacaaagccc ctttcaagaa gatttttgaa
     1921 aagaaaaaaa actaccagat gaaccagaaa aagaagaaga agtatatctt ccccagcatc
     1981 aacatgaacc tggccctgac cagcctgatc cggaagtccg gcctgggcaa cctgcagagc
     2041 tacttcgtgt acgacctggg cttcctgacc ttcgtgttcg gcctgctgaa cgaccgggac
     2101 acccctgagg tgcagcagca gggcgcccac gtgccattct tcaagttcca gcccaagatt
     2161 aacctgaagc tgaacttcaa aggcgcgccg tcgacctcca tcacggccta taagagtgag
     2221 ggggagtcag cggagttctc cttcccactc aaccttggag aggaaagcct gcagggagag
     2281 ttgagatgga aggcagagaa ggctccttct tcccagtcct ggatcacctt ctccctaaag
     2341 aaccaaaagg tgtctgtgca gaagtctact agcaacccca agttccagct gtccgaaacg
     2401 ctcccactca cccttcagat accccaggtc tcccttcagt ttgctggttc tggcaacctg
     2461 accctgactc tggacagagg gatactgtat caggaagtga acctggtggt gatgaaagtg
     2521 actcagcccg acagcaacac tttgacctgt gaggtgatgg gacccacctc acccaagatg
     2581 agactgatct tgaagcagga gaatcaggag gccagggtct ccaggcagga gaaagtgatt
     2641 caagtgcagg cccctgaagc aggggtgtgg caatgtctac tgagtgaagg tgaagaggtc
     2701 aagatggact ccaagatcca ggttttatcc aaagggttga attccggcgg cggctccgga
     2761 ggcggtaccg gtggcggcga cctagcccca cagatgcttc gagaactcca ggagactaat
     2821 gcggcgctgc aagacgtgag agagctcttg cgacagcagg tcaaggagat caccttcctg
     2881 aagaatacgg tgatggaatg tgacgcctgc ggtctagaca gaaacctgcc tccactagca
     2941 cctctgggcc cacacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt
     3001 gcacgagtgg gttacatcga actggatctc aacagcggta agatccttga gagttttcgc
     3061 cccgaagaac gttttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta
     3121 tcccgtattg acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac
     3181 ttggttgagt actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa
     3241 ttatgcagtg ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg
     3301 atcggaggac cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc
     3361 cttgatcgtt gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg
     3421 atgcctgtag caatggcaac aacgttgcgc aaactattaa ctggcgaact acttactcta
     3481 gcttcccggc aacaattaat agactggatg gaggcggata aagttgcagg accacttctg
     3541 cgctcggccc ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggg
     3601 tctcgcggta tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc
     3661 tacacgacgg ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt
     3721 gcctcactga ttaagcattg gggccctgac tacaaagacc atgacggtga ttataaagat
     3781 catgacatcg attacaagga tgacgatgac aagggccctc accaccacca tcaccactaa
     3841 taataaggat cccccgacct cgacctctgg ctaataaagg aaatttattt tcattgcaat
     3901 agtgtgttgg aattttttgt gtctctcact cggaaggaca tatgggaggg caaatcattt
     3961 ggtcgagatc cctcggagat ctctagctag aggatcgatc cccgccccgg acgaactaaa
     4021 cctgactacg acatctctgc cccttcttcg cggggcagtg catgtaatcc cttcagttgg
     4081 ttggtacaac ttgccaactg ggccctgttc cacatgtgac acgggggggg accaaacaca
     4141 aaggggttct ctgactgtag ttgacatcct tataaatgga tgtgcacatt tgccaacact
     4201 gagtggcttt catcctggag cagactttgc agtctgtgga ctgcaacaca acattgcctt
     4261 tatgtgtaac tcttggctga agctcttaca ccaatgctgg gggacatgta cctcccaggg
     4321 gcccaggaag actacgggag gctacaccaa cgtcaatcag aggggcctgt gtagctaccg
     4381 ataagcggac cctcaagagg gcattagcaa tagtgtttat aaggccccct tgttaaccct
     4441 aaacgggtag catatgcttc ccgggtagta gtatatacta tccagactaa ccctaattca
     4501 atagcatatg ttacccaacg ggaagcatat gctatcgaat tagggttagt aaaagggtcc
     4561 taaggaacag cgatatctcc caccccatga gctgtcacgg ttttatttac atggggtcag
     4621 gattccacga gggtagtgaa ccattttagt cacaagggca gtggctgaag atcaaggagc
     4681 gggcagtgaa ctctcctgaa tcttcgcctg cttcttcatt ctccttcgtt tagctaatag
     4741 aataactgct gagttgtgaa cagtaaggtg tatgtgaggt gctcgaaaac aaggtttcag
     4801 gtgacgcccc cagaataaaa tttggacggg gggttcagtg gtggcattgt gctatgacac
     4861 caatataacc ctcacaaacc ccttgggcaa taaatactag tgtaggaatg aaacattctg
     4921 aatatcttta acaatagaaa tccatggggt ggggacaagc cgtaaagact ggatgtccat
     4981 ctcacacgaa tttatggcta tgggcaacac ataatcctag tgcaatatga tactggggtt
     5041 attaagatgt gtcccaggca gggaccaaga caggtgaacc atgttgttac actctatttg
     5101 taacaagggg aaagagagtg gacgccgaca gcagcggact ccactggttg tctctaacac
     5161 ccccgaaaat taaacggggc tccacgccaa tggggcccat aaacaaagac aagtggccac
     5221 tctttttttt gaaattgtgg agtgggggca cgcgtcagcc cccacacgcc gccctgcggt
     5281 tttggactgt aaaataaggg tgtaataact tggctgattg taaccccgct aaccactgcg
     5341 gtcaaaccac ttgcccacaa aaccactaat ggcaccccgg ggaatacctg cataagtagg
     5401 tgggcgggcc aagatagggg cgcgattgct gcgatctgga ggacaaatta cacacacttg
     5461 cgcctgagcg ccaagcacag ggttgttggt cctcatattc acgaggtcgc tgagagcacg
     5521 gtgggctaat gttgccatgg gtagcatata ctacccaaat atctggatag catatgctat
     5581 cctaatctat atctgggtag cataggctat cctaatctat atctgggtag catatgctat
     5641 cctaatctat atctgggtag tatatgctat cctaatttat atctgggtag cataggctat
     5701 cctaatctat atctgggtag catatgctat cctaatctat atctgggtag tatatgctat
     5761 cctaatctgt atccgggtag catatgctat cctaatagag attagggtag tatatgctat
     5821 cctaatttat atctgggtag catatactac ccaaatatct ggatagcata tgctatccta
     5881 atctatatct gggtagcata tgctatccta atctatatct gggtagcata ggctatccta
     5941 atctatatct gggtagcata tgctatccta atctatatct gggtagtata tgctatccta
     6001 atttatatct gggtagcata ggctatccta atctatatct gggtagcata tgctatccta
     6061 atctatatct gggtagtata tgctatccta atctgtatcc gggtagcata tgctatcctc
     6121 atgataagct gtcaaacatg agaattttct tgaagacgaa agggcctcgt gatacgccta
     6181 tttttatagg ttaatgtcat gataataatg gtttcttaga cgtcaggtgg cacttttcgg
     6241 ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg
     6301 ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtatgagt
     6361 attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt
     6421 gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg
     6481 ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa
     6541 cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtgtt
     6601 gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag
     6661 tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt
     6721 gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac gatcggagga
     6781 ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt
     6841 tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgca
     6901 gcaatggcaa caacgttgcg caaactatta actggcgaac tacttactct agcttcccgg
     6961 caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc
     7021 cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt
     7081 atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg
     7141 gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg
     7201 attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat tgatttaaaa
     7261 cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa
     7321 atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga
     7381 tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg
     7441 ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact
     7501 ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac
     7561 cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg
     7621 gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg
     7681 gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga
     7741 acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc
     7801 gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg
     7861 agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc
     7921 tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc
     7981 agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt
     8041 cctgcgttat cccctgattc tgtggataac cgtattaccg cctttgagtg agctgatacc
     8101 gctcgccgca gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc ggaagagcgc
     8161 ccaatacgca aaccgcctct ccccgcgcgt tggccgattc attaatgcag ctggcacgac
     8221 aggtttcccg actggaaagc gggcagtgag cgcaacgcaa ttaatgtgag ttagctcact
     8281 cattaggcac cccaggcttt acactttatg cttccggctc gtatgttgtg tggaattgtg
     8341 agcggataac aatttcacac aggaaacagc tatgaccatg attacgccaa gctctagcta
     8401 gaggtcgacc aattctcatg tttgacagct tatcatcgca gatccgggca acgttgttgc
     8461 cattgctgca ggcgcagaac tggtaggtat ggaagatcta tacattgaat caatattggc
     8521 aattagccat attagtcatt ggttatatag cataaatcaa tattggctat tggccattgc
     8581 atacgttgta tctatatcat aatatgtaca tttatattgg ctcatgtcca atatgaccgc
     8641 catgttgaca ttgattattg actagttatt aatagtaatc aattacgggg tcattagttc
     8701 atagcccata tatggagttc cgcgttacat aacttacggt aaatggcccg cctggctgac
     8761 cgcccaacga cccccgccca ttgacgtcaa taatgacgta tgttcccata gtaacgccaa
     8821 tagggacttt ccattgacgt caatgggtgg agtatttacg gtaaactgcc cacttggcag
     8881 tacatcaagt gtatcatatg ccaagtccgc cccctattga cgtcaatgac ggtaaatggc
     8941 ccgcctggca ttatgcccag tacatgacct tacgggactt tcctacttgg cagtacatct
     9001 acgtattagt catcgctatt accatggtga tgcggttttg gcagtacacc aatgggcgtg
     9061 gatagcggtt tgactcacgg ggatttccaa gtctccaccc cattgacgtc aatgggagtt
     9121 tgttttggca ccaaaatcaa cgggactttc caaaatgtcg taataacccc gccccgttga
     9181 cgcaaatggg cggtaggcgt gtacggtggg aggtctatat aagcagagct cgtttagtga
     9241 accgtcagat cctcactctc ttccgcatcg ctgtctgcga gggccagctg ttgggctcgc
     9301 ggttgaggac aaactcttcg cggtctttcc agtactcttg gatcggaaac ccgtcggcct
     9361 ccgaacggta ctccgccacc gagggacctg agcgagtccg catcgaccgg atcggaaaac
     9421 ct
//
