LOCUS       sequence-224578-        8342 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 8342)
  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 8342)
  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: GEST-COMP-blac-flag-his
FEATURES             Location/Qualifiers
     source          1..8342
                     /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             1876..2661
                     /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             2668..2733
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             2740..2757
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    2793..2848
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(2847..2866)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(5084..5102)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5170..5274
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5275..5343,5344..6135)
                     /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      6306..6894
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6795..6814
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7048..7065
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    7182..7203
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7218..7223,7224..7241,7242..7248)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7256..7272
                     /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     7261..7283
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7280..7296
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7280..7296
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     enhancer        7567..7946
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        7947..8150
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     8101..8121
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     8147..8171
                     /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 ggccgccatt
      421 cagctgcaca gcgtgaacta cgcccctctg aactacatcg agatcggcga caccctgagc
      481 aacaaagtgg gcgagctgtg gcacagccac ctgaacagct tcatcgacat cgtgtccacc
      541 aaggtggtca acaagctgga agatgacctg tccaacggca acaacctcga gaagttcatg
      601 atcctgctcg aggacgacgc cgacatcttc gacgccagcg cctacgaggg caagaacctg
      661 gccctgatcc agaccgagtt catccggaag ctgaaggaca agttcaagaa cagcaagttc
      721 ggccagaagc tgaaaaagct gggcagcaag gccaaagaga agctgctgag cctgtaccag
      781 aagcacaagg gcaagctgcg gcacttcttc agcatcctgc tgaagtccct ggtcatccct
      841 atcgccgtgc agttcatcag aaagaacctg aacaagtgga agcagcggac cctggaagcc
      901 acacagaagc tggacgagca gagcaagaat atcgcccagc ccatcatcaa ccggctgtac
      961 aacagcttcg aggacaagat cgaagagtac agccaggaaa acaagatcaa cgtcgaggat
     1021 gagctgaacg ccctgggcca gctggacaag gacaagcagg acatccagaa actcgaggaa
     1081 caggaaaagg ctctgctgca gggcgcgccg tcgacctcca tcacggccta taagagtgag
     1141 ggggagtcag cggagttctc cttcccactc aaccttggag aggaaagcct gcagggagag
     1201 ttgagatgga aggcagagaa ggctccttct tcccagtcct ggatcacctt ctccctaaag
     1261 aaccaaaagg tgtctgtgca gaagtctact agcaacccca agttccagct gtccgaaacg
     1321 ctcccactca cccttcagat accccaggtc tcccttcagt ttgctggttc tggcaacctg
     1381 accctgactc tggacagagg gatactgtat caggaagtga acctggtggt gatgaaagtg
     1441 actcagcccg acagcaacac tttgacctgt gaggtgatgg gacccacctc acccaagatg
     1501 agactgatct tgaagcagga gaatcaggag gccagggtct ccaggcagga gaaagtgatt
     1561 caagtgcagg cccctgaagc aggggtgtgg caatgtctac tgagtgaagg tgaagaggtc
     1621 aagatggact ccaagatcca ggttttatcc aaagggttga attccggcgg cggctccgga
     1681 ggcggtaccg gtggcggcga cctagcccca cagatgcttc gagaactcca ggagactaat
     1741 gcggcgctgc aagacgtgag agagctcttg cgacagcagg tcaaggagat caccttcctg
     1801 aagaatacgg tgatggaatg tgacgcctgc ggtctagaca gaaacctgcc tccactagca
     1861 cctctgggcc cacacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt
     1921 gcacgagtgg gttacatcga actggatctc aacagcggta agatccttga gagttttcgc
     1981 cccgaagaac gttttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta
     2041 tcccgtattg acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac
     2101 ttggttgagt actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa
     2161 ttatgcagtg ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg
     2221 atcggaggac cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc
     2281 cttgatcgtt gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg
     2341 atgcctgtag caatggcaac aacgttgcgc aaactattaa ctggcgaact acttactcta
     2401 gcttcccggc aacaattaat agactggatg gaggcggata aagttgcagg accacttctg
     2461 cgctcggccc ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggg
     2521 tctcgcggta tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc
     2581 tacacgacgg ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt
     2641 gcctcactga ttaagcattg gggccctgac tacaaagacc atgacggtga ttataaagat
     2701 catgacatcg attacaagga tgacgatgac aagggccctc accaccacca tcaccactaa
     2761 taataaggat cccccgacct cgacctctgg ctaataaagg aaatttattt tcattgcaat
     2821 agtgtgttgg aattttttgt gtctctcact cggaaggaca tatgggaggg caaatcattt
     2881 ggtcgagatc cctcggagat ctctagctag aggatcgatc cccgccccgg acgaactaaa
     2941 cctgactacg acatctctgc cccttcttcg cggggcagtg catgtaatcc cttcagttgg
     3001 ttggtacaac ttgccaactg ggccctgttc cacatgtgac acgggggggg accaaacaca
     3061 aaggggttct ctgactgtag ttgacatcct tataaatgga tgtgcacatt tgccaacact
     3121 gagtggcttt catcctggag cagactttgc agtctgtgga ctgcaacaca acattgcctt
     3181 tatgtgtaac tcttggctga agctcttaca ccaatgctgg gggacatgta cctcccaggg
     3241 gcccaggaag actacgggag gctacaccaa cgtcaatcag aggggcctgt gtagctaccg
     3301 ataagcggac cctcaagagg gcattagcaa tagtgtttat aaggccccct tgttaaccct
     3361 aaacgggtag catatgcttc ccgggtagta gtatatacta tccagactaa ccctaattca
     3421 atagcatatg ttacccaacg ggaagcatat gctatcgaat tagggttagt aaaagggtcc
     3481 taaggaacag cgatatctcc caccccatga gctgtcacgg ttttatttac atggggtcag
     3541 gattccacga gggtagtgaa ccattttagt cacaagggca gtggctgaag atcaaggagc
     3601 gggcagtgaa ctctcctgaa tcttcgcctg cttcttcatt ctccttcgtt tagctaatag
     3661 aataactgct gagttgtgaa cagtaaggtg tatgtgaggt gctcgaaaac aaggtttcag
     3721 gtgacgcccc cagaataaaa tttggacggg gggttcagtg gtggcattgt gctatgacac
     3781 caatataacc ctcacaaacc ccttgggcaa taaatactag tgtaggaatg aaacattctg
     3841 aatatcttta acaatagaaa tccatggggt ggggacaagc cgtaaagact ggatgtccat
     3901 ctcacacgaa tttatggcta tgggcaacac ataatcctag tgcaatatga tactggggtt
     3961 attaagatgt gtcccaggca gggaccaaga caggtgaacc atgttgttac actctatttg
     4021 taacaagggg aaagagagtg gacgccgaca gcagcggact ccactggttg tctctaacac
     4081 ccccgaaaat taaacggggc tccacgccaa tggggcccat aaacaaagac aagtggccac
     4141 tctttttttt gaaattgtgg agtgggggca cgcgtcagcc cccacacgcc gccctgcggt
     4201 tttggactgt aaaataaggg tgtaataact tggctgattg taaccccgct aaccactgcg
     4261 gtcaaaccac ttgcccacaa aaccactaat ggcaccccgg ggaatacctg cataagtagg
     4321 tgggcgggcc aagatagggg cgcgattgct gcgatctgga ggacaaatta cacacacttg
     4381 cgcctgagcg ccaagcacag ggttgttggt cctcatattc acgaggtcgc tgagagcacg
     4441 gtgggctaat gttgccatgg gtagcatata ctacccaaat atctggatag catatgctat
     4501 cctaatctat atctgggtag cataggctat cctaatctat atctgggtag catatgctat
     4561 cctaatctat atctgggtag tatatgctat cctaatttat atctgggtag cataggctat
     4621 cctaatctat atctgggtag catatgctat cctaatctat atctgggtag tatatgctat
     4681 cctaatctgt atccgggtag catatgctat cctaatagag attagggtag tatatgctat
     4741 cctaatttat atctgggtag catatactac ccaaatatct ggatagcata tgctatccta
     4801 atctatatct gggtagcata tgctatccta atctatatct gggtagcata ggctatccta
     4861 atctatatct gggtagcata tgctatccta atctatatct gggtagtata tgctatccta
     4921 atttatatct gggtagcata ggctatccta atctatatct gggtagcata tgctatccta
     4981 atctatatct gggtagtata tgctatccta atctgtatcc gggtagcata tgctatcctc
     5041 atgataagct gtcaaacatg agaattttct tgaagacgaa agggcctcgt gatacgccta
     5101 tttttatagg ttaatgtcat gataataatg gtttcttaga cgtcaggtgg cacttttcgg
     5161 ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg
     5221 ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtatgagt
     5281 attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt
     5341 gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg
     5401 ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa
     5461 cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtgtt
     5521 gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag
     5581 tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt
     5641 gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac gatcggagga
     5701 ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt
     5761 tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgca
     5821 gcaatggcaa caacgttgcg caaactatta actggcgaac tacttactct agcttcccgg
     5881 caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc
     5941 cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt
     6001 atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg
     6061 gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg
     6121 attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat tgatttaaaa
     6181 cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa
     6241 atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga
     6301 tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg
     6361 ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact
     6421 ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac
     6481 cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg
     6541 gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg
     6601 gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga
     6661 acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc
     6721 gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg
     6781 agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc
     6841 tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc
     6901 agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt
     6961 cctgcgttat cccctgattc tgtggataac cgtattaccg cctttgagtg agctgatacc
     7021 gctcgccgca gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc ggaagagcgc
     7081 ccaatacgca aaccgcctct ccccgcgcgt tggccgattc attaatgcag ctggcacgac
     7141 aggtttcccg actggaaagc gggcagtgag cgcaacgcaa ttaatgtgag ttagctcact
     7201 cattaggcac cccaggcttt acactttatg cttccggctc gtatgttgtg tggaattgtg
     7261 agcggataac aatttcacac aggaaacagc tatgaccatg attacgccaa gctctagcta
     7321 gaggtcgacc aattctcatg tttgacagct tatcatcgca gatccgggca acgttgttgc
     7381 cattgctgca ggcgcagaac tggtaggtat ggaagatcta tacattgaat caatattggc
     7441 aattagccat attagtcatt ggttatatag cataaatcaa tattggctat tggccattgc
     7501 atacgttgta tctatatcat aatatgtaca tttatattgg ctcatgtcca atatgaccgc
     7561 catgttgaca ttgattattg actagttatt aatagtaatc aattacgggg tcattagttc
     7621 atagcccata tatggagttc cgcgttacat aacttacggt aaatggcccg cctggctgac
     7681 cgcccaacga cccccgccca ttgacgtcaa taatgacgta tgttcccata gtaacgccaa
     7741 tagggacttt ccattgacgt caatgggtgg agtatttacg gtaaactgcc cacttggcag
     7801 tacatcaagt gtatcatatg ccaagtccgc cccctattga cgtcaatgac ggtaaatggc
     7861 ccgcctggca ttatgcccag tacatgacct tacgggactt tcctacttgg cagtacatct
     7921 acgtattagt catcgctatt accatggtga tgcggttttg gcagtacacc aatgggcgtg
     7981 gatagcggtt tgactcacgg ggatttccaa gtctccaccc cattgacgtc aatgggagtt
     8041 tgttttggca ccaaaatcaa cgggactttc caaaatgtcg taataacccc gccccgttga
     8101 cgcaaatggg cggtaggcgt gtacggtggg aggtctatat aagcagagct cgtttagtga
     8161 accgtcagat cctcactctc ttccgcatcg ctgtctgcga gggccagctg ttgggctcgc
     8221 ggttgaggac aaactcttcg cggtctttcc agtactcttg gatcggaaac ccgtcggcct
     8281 ccgaacggta ctccgccacc gagggacctg agcgagtccg catcgaccgg atcggaaaac
     8341 ct
//
