LOCUS       sequence-224251-        8102 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 8102)
  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 8102)
  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_1352500-COMP-blac-flag-his
FEATURES             Location/Qualifiers
     source          1..8102
                     /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             1636..2421
                     /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             2428..2493
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             2500..2517
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    2553..2608
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(2607..2626)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(4844..4862)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4930..5034
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5035..5103,5104..5895)
                     /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      6066..6654
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6555..6574
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6808..6825
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    6942..6963
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6978..6983,6984..7001,7002..7008)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7016..7032
                     /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     7021..7043
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7040..7056
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7040..7056
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     enhancer        7327..7706
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        7707..7910
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     7861..7881
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     7907..7931
                     /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 ggccgcccag
      421 gacgtgatcg agctgaacga cgccaacttc gagagcctga cccagctgag caccggcaac
      481 accgccggca gctggttcat caagttctac gccccttggt gcagccactg caaggccatg
      541 agcaagacat gggcccagct ggccaccgag ctgaagggca agatcaacgt ggccaagatc
      601 gacgtgaccc tgaacagcaa gacccggaag cggttcaaga tcgagggctt ccccaccctg
      661 ctgtacttca agaacggcaa gatgtacgac tacaagaacc acgacagatc cctcgaggcc
      721 ttcaagaatt tcgtgctcga gacatacaag aacgccaagg ccagcgagcc tcccaagccc
      781 ctgaactaca tggatatcct gaaggacttc ctgaacgagg cctttcagaa catcgaccgg
      841 atctataagt acgccttccc cggcgcgccg tcgacctcca tcacggccta taagagtgag
      901 ggggagtcag cggagttctc cttcccactc aaccttggag aggaaagcct gcagggagag
      961 ttgagatgga aggcagagaa ggctccttct tcccagtcct ggatcacctt ctccctaaag
     1021 aaccaaaagg tgtctgtgca gaagtctact agcaacccca agttccagct gtccgaaacg
     1081 ctcccactca cccttcagat accccaggtc tcccttcagt ttgctggttc tggcaacctg
     1141 accctgactc tggacagagg gatactgtat caggaagtga acctggtggt gatgaaagtg
     1201 actcagcccg acagcaacac tttgacctgt gaggtgatgg gacccacctc acccaagatg
     1261 agactgatct tgaagcagga gaatcaggag gccagggtct ccaggcagga gaaagtgatt
     1321 caagtgcagg cccctgaagc aggggtgtgg caatgtctac tgagtgaagg tgaagaggtc
     1381 aagatggact ccaagatcca ggttttatcc aaagggttga attccggcgg cggctccgga
     1441 ggcggtaccg gtggcggcga cctagcccca cagatgcttc gagaactcca ggagactaat
     1501 gcggcgctgc aagacgtgag agagctcttg cgacagcagg tcaaggagat caccttcctg
     1561 aagaatacgg tgatggaatg tgacgcctgc ggtctagaca gaaacctgcc tccactagca
     1621 cctctgggcc cacacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt
     1681 gcacgagtgg gttacatcga actggatctc aacagcggta agatccttga gagttttcgc
     1741 cccgaagaac gttttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta
     1801 tcccgtattg acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac
     1861 ttggttgagt actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa
     1921 ttatgcagtg ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg
     1981 atcggaggac cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc
     2041 cttgatcgtt gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg
     2101 atgcctgtag caatggcaac aacgttgcgc aaactattaa ctggcgaact acttactcta
     2161 gcttcccggc aacaattaat agactggatg gaggcggata aagttgcagg accacttctg
     2221 cgctcggccc ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggg
     2281 tctcgcggta tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc
     2341 tacacgacgg ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt
     2401 gcctcactga ttaagcattg gggccctgac tacaaagacc atgacggtga ttataaagat
     2461 catgacatcg attacaagga tgacgatgac aagggccctc accaccacca tcaccactaa
     2521 taataaggat cccccgacct cgacctctgg ctaataaagg aaatttattt tcattgcaat
     2581 agtgtgttgg aattttttgt gtctctcact cggaaggaca tatgggaggg caaatcattt
     2641 ggtcgagatc cctcggagat ctctagctag aggatcgatc cccgccccgg acgaactaaa
     2701 cctgactacg acatctctgc cccttcttcg cggggcagtg catgtaatcc cttcagttgg
     2761 ttggtacaac ttgccaactg ggccctgttc cacatgtgac acgggggggg accaaacaca
     2821 aaggggttct ctgactgtag ttgacatcct tataaatgga tgtgcacatt tgccaacact
     2881 gagtggcttt catcctggag cagactttgc agtctgtgga ctgcaacaca acattgcctt
     2941 tatgtgtaac tcttggctga agctcttaca ccaatgctgg gggacatgta cctcccaggg
     3001 gcccaggaag actacgggag gctacaccaa cgtcaatcag aggggcctgt gtagctaccg
     3061 ataagcggac cctcaagagg gcattagcaa tagtgtttat aaggccccct tgttaaccct
     3121 aaacgggtag catatgcttc ccgggtagta gtatatacta tccagactaa ccctaattca
     3181 atagcatatg ttacccaacg ggaagcatat gctatcgaat tagggttagt aaaagggtcc
     3241 taaggaacag cgatatctcc caccccatga gctgtcacgg ttttatttac atggggtcag
     3301 gattccacga gggtagtgaa ccattttagt cacaagggca gtggctgaag atcaaggagc
     3361 gggcagtgaa ctctcctgaa tcttcgcctg cttcttcatt ctccttcgtt tagctaatag
     3421 aataactgct gagttgtgaa cagtaaggtg tatgtgaggt gctcgaaaac aaggtttcag
     3481 gtgacgcccc cagaataaaa tttggacggg gggttcagtg gtggcattgt gctatgacac
     3541 caatataacc ctcacaaacc ccttgggcaa taaatactag tgtaggaatg aaacattctg
     3601 aatatcttta acaatagaaa tccatggggt ggggacaagc cgtaaagact ggatgtccat
     3661 ctcacacgaa tttatggcta tgggcaacac ataatcctag tgcaatatga tactggggtt
     3721 attaagatgt gtcccaggca gggaccaaga caggtgaacc atgttgttac actctatttg
     3781 taacaagggg aaagagagtg gacgccgaca gcagcggact ccactggttg tctctaacac
     3841 ccccgaaaat taaacggggc tccacgccaa tggggcccat aaacaaagac aagtggccac
     3901 tctttttttt gaaattgtgg agtgggggca cgcgtcagcc cccacacgcc gccctgcggt
     3961 tttggactgt aaaataaggg tgtaataact tggctgattg taaccccgct aaccactgcg
     4021 gtcaaaccac ttgcccacaa aaccactaat ggcaccccgg ggaatacctg cataagtagg
     4081 tgggcgggcc aagatagggg cgcgattgct gcgatctgga ggacaaatta cacacacttg
     4141 cgcctgagcg ccaagcacag ggttgttggt cctcatattc acgaggtcgc tgagagcacg
     4201 gtgggctaat gttgccatgg gtagcatata ctacccaaat atctggatag catatgctat
     4261 cctaatctat atctgggtag cataggctat cctaatctat atctgggtag catatgctat
     4321 cctaatctat atctgggtag tatatgctat cctaatttat atctgggtag cataggctat
     4381 cctaatctat atctgggtag catatgctat cctaatctat atctgggtag tatatgctat
     4441 cctaatctgt atccgggtag catatgctat cctaatagag attagggtag tatatgctat
     4501 cctaatttat atctgggtag catatactac ccaaatatct ggatagcata tgctatccta
     4561 atctatatct gggtagcata tgctatccta atctatatct gggtagcata ggctatccta
     4621 atctatatct gggtagcata tgctatccta atctatatct gggtagtata tgctatccta
     4681 atttatatct gggtagcata ggctatccta atctatatct gggtagcata tgctatccta
     4741 atctatatct gggtagtata tgctatccta atctgtatcc gggtagcata tgctatcctc
     4801 atgataagct gtcaaacatg agaattttct tgaagacgaa agggcctcgt gatacgccta
     4861 tttttatagg ttaatgtcat gataataatg gtttcttaga cgtcaggtgg cacttttcgg
     4921 ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg
     4981 ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtatgagt
     5041 attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt
     5101 gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg
     5161 ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa
     5221 cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtgtt
     5281 gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag
     5341 tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt
     5401 gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac gatcggagga
     5461 ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt
     5521 tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgca
     5581 gcaatggcaa caacgttgcg caaactatta actggcgaac tacttactct agcttcccgg
     5641 caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc
     5701 cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt
     5761 atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg
     5821 gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg
     5881 attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat tgatttaaaa
     5941 cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa
     6001 atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga
     6061 tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg
     6121 ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact
     6181 ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac
     6241 cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg
     6301 gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg
     6361 gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga
     6421 acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc
     6481 gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg
     6541 agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc
     6601 tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc
     6661 agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt
     6721 cctgcgttat cccctgattc tgtggataac cgtattaccg cctttgagtg agctgatacc
     6781 gctcgccgca gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc ggaagagcgc
     6841 ccaatacgca aaccgcctct ccccgcgcgt tggccgattc attaatgcag ctggcacgac
     6901 aggtttcccg actggaaagc gggcagtgag cgcaacgcaa ttaatgtgag ttagctcact
     6961 cattaggcac cccaggcttt acactttatg cttccggctc gtatgttgtg tggaattgtg
     7021 agcggataac aatttcacac aggaaacagc tatgaccatg attacgccaa gctctagcta
     7081 gaggtcgacc aattctcatg tttgacagct tatcatcgca gatccgggca acgttgttgc
     7141 cattgctgca ggcgcagaac tggtaggtat ggaagatcta tacattgaat caatattggc
     7201 aattagccat attagtcatt ggttatatag cataaatcaa tattggctat tggccattgc
     7261 atacgttgta tctatatcat aatatgtaca tttatattgg ctcatgtcca atatgaccgc
     7321 catgttgaca ttgattattg actagttatt aatagtaatc aattacgggg tcattagttc
     7381 atagcccata tatggagttc cgcgttacat aacttacggt aaatggcccg cctggctgac
     7441 cgcccaacga cccccgccca ttgacgtcaa taatgacgta tgttcccata gtaacgccaa
     7501 tagggacttt ccattgacgt caatgggtgg agtatttacg gtaaactgcc cacttggcag
     7561 tacatcaagt gtatcatatg ccaagtccgc cccctattga cgtcaatgac ggtaaatggc
     7621 ccgcctggca ttatgcccag tacatgacct tacgggactt tcctacttgg cagtacatct
     7681 acgtattagt catcgctatt accatggtga tgcggttttg gcagtacacc aatgggcgtg
     7741 gatagcggtt tgactcacgg ggatttccaa gtctccaccc cattgacgtc aatgggagtt
     7801 tgttttggca ccaaaatcaa cgggactttc caaaatgtcg taataacccc gccccgttga
     7861 cgcaaatggg cggtaggcgt gtacggtggg aggtctatat aagcagagct cgtttagtga
     7921 accgtcagat cctcactctc ttccgcatcg ctgtctgcga gggccagctg ttgggctcgc
     7981 ggttgaggac aaactcttcg cggtctttcc agtactcttg gatcggaaac ccgtcggcct
     8041 ccgaacggta ctccgccacc gagggacctg agcgagtccg catcgaccgg atcggaaaac
     8101 ct
//
