LOCUS       sequence-224371-        8642 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 8642)
  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 8642)
  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_0422100-COMP-blac-flag-his
FEATURES             Location/Qualifiers
     source          1..8642
                     /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             2176..2961
                     /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             2968..3033
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             3040..3057
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    3093..3148
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(3147..3166)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(5384..5402)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5470..5574
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5575..5643,5644..6435)
                     /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      6606..7194
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7095..7114
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7348..7365
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    7482..7503
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7518..7523,7524..7541,7542..7548)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7556..7572
                     /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     7561..7583
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7580..7596
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7580..7596
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     enhancer        7867..8246
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        8247..8450
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     8401..8421
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     8447..8471
                     /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 ggccgccaac
      421 ttcgtgctga gcaacgccaa caccaacaac gtgaacaaga acggcgccaa caagaagaac
      481 aacgctaaca acaataatat gaacaaggcc ggcgataaga agaaggccac caagggcaag
      541 aagggaaacg ccaacgacaa tgtgaacgag ttcaacttca atttcaacga cctgctcaac
      601 gagatcaaca acgccgccac caacgataac aataacaaca acaatgacaa taataacgac
      661 aacaacaaca acaataacaa caacaacaac aattacaatg gcaagaacga gatcaatgag
      721 aacaacttca acgaagagta caacaagctc gagaagaact tccagatgaa cgccgaggac
      781 gatgacaaca accggggcga catcaacatg gaccagttca acgagaagaa aaacaacctg
      841 aacgaggata ataacaagat gaagcccaac gccaatgact accagtataa cgagaacgac
      901 gccgacaccg acagcgaaga ggacctgacc aacatctgga acaagaatat gcagaacttc
      961 gagcccagca ccctgctgac cttcgagatc cccagcaaca gcgaggaata cctgttcgaa
     1021 gaggtgcccg aagtgaacac ctacttccgg ggcgtgttct actccaacaa cgaggccgac
     1081 gacaacatca tcgagtttct gatcagcgac cccgacggcc aggtcatcta caagaaagag
     1141 gccagcgagg gcatcttcta cttctacacc aagaaagtgg gcatctacac cctgaccctc
     1201 aagaacaaca agtggatggg caagaaactg accaccgtgg ccctgggcct gggcgagaat
     1261 cccgccctga gaagcgacca cgtgaaggac ttcaccaact acatccagcg gatcgtggcc
     1321 gagacaaaga agctgaaaaa cgagattaag tacctgagca gcaagcacat gcggcacatc
     1381 gagaagatga agaagatcac cggcgcgccg tcgacctcca tcacggccta taagagtgag
     1441 ggggagtcag cggagttctc cttcccactc aaccttggag aggaaagcct gcagggagag
     1501 ttgagatgga aggcagagaa ggctccttct tcccagtcct ggatcacctt ctccctaaag
     1561 aaccaaaagg tgtctgtgca gaagtctact agcaacccca agttccagct gtccgaaacg
     1621 ctcccactca cccttcagat accccaggtc tcccttcagt ttgctggttc tggcaacctg
     1681 accctgactc tggacagagg gatactgtat caggaagtga acctggtggt gatgaaagtg
     1741 actcagcccg acagcaacac tttgacctgt gaggtgatgg gacccacctc acccaagatg
     1801 agactgatct tgaagcagga gaatcaggag gccagggtct ccaggcagga gaaagtgatt
     1861 caagtgcagg cccctgaagc aggggtgtgg caatgtctac tgagtgaagg tgaagaggtc
     1921 aagatggact ccaagatcca ggttttatcc aaagggttga attccggcgg cggctccgga
     1981 ggcggtaccg gtggcggcga cctagcccca cagatgcttc gagaactcca ggagactaat
     2041 gcggcgctgc aagacgtgag agagctcttg cgacagcagg tcaaggagat caccttcctg
     2101 aagaatacgg tgatggaatg tgacgcctgc ggtctagaca gaaacctgcc tccactagca
     2161 cctctgggcc cacacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt
     2221 gcacgagtgg gttacatcga actggatctc aacagcggta agatccttga gagttttcgc
     2281 cccgaagaac gttttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta
     2341 tcccgtattg acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac
     2401 ttggttgagt actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa
     2461 ttatgcagtg ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg
     2521 atcggaggac cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc
     2581 cttgatcgtt gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg
     2641 atgcctgtag caatggcaac aacgttgcgc aaactattaa ctggcgaact acttactcta
     2701 gcttcccggc aacaattaat agactggatg gaggcggata aagttgcagg accacttctg
     2761 cgctcggccc ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggg
     2821 tctcgcggta tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc
     2881 tacacgacgg ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt
     2941 gcctcactga ttaagcattg gggccctgac tacaaagacc atgacggtga ttataaagat
     3001 catgacatcg attacaagga tgacgatgac aagggccctc accaccacca tcaccactaa
     3061 taataaggat cccccgacct cgacctctgg ctaataaagg aaatttattt tcattgcaat
     3121 agtgtgttgg aattttttgt gtctctcact cggaaggaca tatgggaggg caaatcattt
     3181 ggtcgagatc cctcggagat ctctagctag aggatcgatc cccgccccgg acgaactaaa
     3241 cctgactacg acatctctgc cccttcttcg cggggcagtg catgtaatcc cttcagttgg
     3301 ttggtacaac ttgccaactg ggccctgttc cacatgtgac acgggggggg accaaacaca
     3361 aaggggttct ctgactgtag ttgacatcct tataaatgga tgtgcacatt tgccaacact
     3421 gagtggcttt catcctggag cagactttgc agtctgtgga ctgcaacaca acattgcctt
     3481 tatgtgtaac tcttggctga agctcttaca ccaatgctgg gggacatgta cctcccaggg
     3541 gcccaggaag actacgggag gctacaccaa cgtcaatcag aggggcctgt gtagctaccg
     3601 ataagcggac cctcaagagg gcattagcaa tagtgtttat aaggccccct tgttaaccct
     3661 aaacgggtag catatgcttc ccgggtagta gtatatacta tccagactaa ccctaattca
     3721 atagcatatg ttacccaacg ggaagcatat gctatcgaat tagggttagt aaaagggtcc
     3781 taaggaacag cgatatctcc caccccatga gctgtcacgg ttttatttac atggggtcag
     3841 gattccacga gggtagtgaa ccattttagt cacaagggca gtggctgaag atcaaggagc
     3901 gggcagtgaa ctctcctgaa tcttcgcctg cttcttcatt ctccttcgtt tagctaatag
     3961 aataactgct gagttgtgaa cagtaaggtg tatgtgaggt gctcgaaaac aaggtttcag
     4021 gtgacgcccc cagaataaaa tttggacggg gggttcagtg gtggcattgt gctatgacac
     4081 caatataacc ctcacaaacc ccttgggcaa taaatactag tgtaggaatg aaacattctg
     4141 aatatcttta acaatagaaa tccatggggt ggggacaagc cgtaaagact ggatgtccat
     4201 ctcacacgaa tttatggcta tgggcaacac ataatcctag tgcaatatga tactggggtt
     4261 attaagatgt gtcccaggca gggaccaaga caggtgaacc atgttgttac actctatttg
     4321 taacaagggg aaagagagtg gacgccgaca gcagcggact ccactggttg tctctaacac
     4381 ccccgaaaat taaacggggc tccacgccaa tggggcccat aaacaaagac aagtggccac
     4441 tctttttttt gaaattgtgg agtgggggca cgcgtcagcc cccacacgcc gccctgcggt
     4501 tttggactgt aaaataaggg tgtaataact tggctgattg taaccccgct aaccactgcg
     4561 gtcaaaccac ttgcccacaa aaccactaat ggcaccccgg ggaatacctg cataagtagg
     4621 tgggcgggcc aagatagggg cgcgattgct gcgatctgga ggacaaatta cacacacttg
     4681 cgcctgagcg ccaagcacag ggttgttggt cctcatattc acgaggtcgc tgagagcacg
     4741 gtgggctaat gttgccatgg gtagcatata ctacccaaat atctggatag catatgctat
     4801 cctaatctat atctgggtag cataggctat cctaatctat atctgggtag catatgctat
     4861 cctaatctat atctgggtag tatatgctat cctaatttat atctgggtag cataggctat
     4921 cctaatctat atctgggtag catatgctat cctaatctat atctgggtag tatatgctat
     4981 cctaatctgt atccgggtag catatgctat cctaatagag attagggtag tatatgctat
     5041 cctaatttat atctgggtag catatactac ccaaatatct ggatagcata tgctatccta
     5101 atctatatct gggtagcata tgctatccta atctatatct gggtagcata ggctatccta
     5161 atctatatct gggtagcata tgctatccta atctatatct gggtagtata tgctatccta
     5221 atttatatct gggtagcata ggctatccta atctatatct gggtagcata tgctatccta
     5281 atctatatct gggtagtata tgctatccta atctgtatcc gggtagcata tgctatcctc
     5341 atgataagct gtcaaacatg agaattttct tgaagacgaa agggcctcgt gatacgccta
     5401 tttttatagg ttaatgtcat gataataatg gtttcttaga cgtcaggtgg cacttttcgg
     5461 ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg
     5521 ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtatgagt
     5581 attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt
     5641 gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg
     5701 ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa
     5761 cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtgtt
     5821 gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag
     5881 tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt
     5941 gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac gatcggagga
     6001 ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt
     6061 tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgca
     6121 gcaatggcaa caacgttgcg caaactatta actggcgaac tacttactct agcttcccgg
     6181 caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc
     6241 cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt
     6301 atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg
     6361 gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg
     6421 attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat tgatttaaaa
     6481 cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa
     6541 atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga
     6601 tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg
     6661 ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact
     6721 ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac
     6781 cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg
     6841 gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg
     6901 gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga
     6961 acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc
     7021 gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg
     7081 agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc
     7141 tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc
     7201 agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt
     7261 cctgcgttat cccctgattc tgtggataac cgtattaccg cctttgagtg agctgatacc
     7321 gctcgccgca gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc ggaagagcgc
     7381 ccaatacgca aaccgcctct ccccgcgcgt tggccgattc attaatgcag ctggcacgac
     7441 aggtttcccg actggaaagc gggcagtgag cgcaacgcaa ttaatgtgag ttagctcact
     7501 cattaggcac cccaggcttt acactttatg cttccggctc gtatgttgtg tggaattgtg
     7561 agcggataac aatttcacac aggaaacagc tatgaccatg attacgccaa gctctagcta
     7621 gaggtcgacc aattctcatg tttgacagct tatcatcgca gatccgggca acgttgttgc
     7681 cattgctgca ggcgcagaac tggtaggtat ggaagatcta tacattgaat caatattggc
     7741 aattagccat attagtcatt ggttatatag cataaatcaa tattggctat tggccattgc
     7801 atacgttgta tctatatcat aatatgtaca tttatattgg ctcatgtcca atatgaccgc
     7861 catgttgaca ttgattattg actagttatt aatagtaatc aattacgggg tcattagttc
     7921 atagcccata tatggagttc cgcgttacat aacttacggt aaatggcccg cctggctgac
     7981 cgcccaacga cccccgccca ttgacgtcaa taatgacgta tgttcccata gtaacgccaa
     8041 tagggacttt ccattgacgt caatgggtgg agtatttacg gtaaactgcc cacttggcag
     8101 tacatcaagt gtatcatatg ccaagtccgc cccctattga cgtcaatgac ggtaaatggc
     8161 ccgcctggca ttatgcccag tacatgacct tacgggactt tcctacttgg cagtacatct
     8221 acgtattagt catcgctatt accatggtga tgcggttttg gcagtacacc aatgggcgtg
     8281 gatagcggtt tgactcacgg ggatttccaa gtctccaccc cattgacgtc aatgggagtt
     8341 tgttttggca ccaaaatcaa cgggactttc caaaatgtcg taataacccc gccccgttga
     8401 cgcaaatggg cggtaggcgt gtacggtggg aggtctatat aagcagagct cgtttagtga
     8461 accgtcagat cctcactctc ttccgcatcg ctgtctgcga gggccagctg ttgggctcgc
     8521 ggttgaggac aaactcttcg cggtctttcc agtactcttg gatcggaaac ccgtcggcct
     8581 ccgaacggta ctccgccacc gagggacctg agcgagtccg catcgaccgg atcggaaaac
     8641 ct
//
