LOCUS       sequence-224577-        8132 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 8132)
  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 8132)
  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: CelTOS-COMP-blac-flag-his
FEATURES             Location/Qualifiers
     source          1..8132
                     /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             1666..2451
                     /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             2458..2523
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             2530..2547
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    2583..2638
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(2637..2656)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(4874..4892)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4960..5064
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5065..5133,5134..5925)
                     /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      6096..6684
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6585..6604
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6838..6855
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    6972..6993
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7008..7013,7014..7031,7032..7038)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7046..7062
                     /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     7051..7073
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7070..7086
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7070..7086
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     enhancer        7357..7736
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        7737..7940
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     7891..7911
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     7937..7961
                     /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 ggccgccttc
      421 agaggcaaca acggccacaa cagcgccagc agcctgtaca acggcgccca gttcatcgag
      481 cagctgaaca acgccttcac cagcgccttt ctggaaagcc agagcatgaa caagatcggc
      541 gacgacctgg ccgagacaat cagcaacgag ctggtgtccg tgctgcagaa gaacagcccc
      601 accttcctgg aatccagctt cgacatcaag agcgaagtga agaaacacgc caagagcatg
      661 ctgaaagagc tgatcaaagt cggcctgcct agcttcgaga acctggtggc cgagaacgtg
      721 aagcctccca aggtggaccc cgccacctac ggcatcatcg tgcccgtgct gaccagcctg
      781 ttcaacaagg tggaaaccgc cgtgggcgcc aaggtgtccg acgagatctg gaactacaac
      841 agccccgacg tgtccgagag cgaggaaagc ctgagcgacg acttcttcga cggcgcgccg
      901 tcgacctcca tcacggccta taagagtgag ggggagtcag cggagttctc cttcccactc
      961 aaccttggag aggaaagcct gcagggagag ttgagatgga aggcagagaa ggctccttct
     1021 tcccagtcct ggatcacctt ctccctaaag aaccaaaagg tgtctgtgca gaagtctact
     1081 agcaacccca agttccagct gtccgaaacg ctcccactca cccttcagat accccaggtc
     1141 tcccttcagt ttgctggttc tggcaacctg accctgactc tggacagagg gatactgtat
     1201 caggaagtga acctggtggt gatgaaagtg actcagcccg acagcaacac tttgacctgt
     1261 gaggtgatgg gacccacctc acccaagatg agactgatct tgaagcagga gaatcaggag
     1321 gccagggtct ccaggcagga gaaagtgatt caagtgcagg cccctgaagc aggggtgtgg
     1381 caatgtctac tgagtgaagg tgaagaggtc aagatggact ccaagatcca ggttttatcc
     1441 aaagggttga attccggcgg cggctccgga ggcggtaccg gtggcggcga cctagcccca
     1501 cagatgcttc gagaactcca ggagactaat gcggcgctgc aagacgtgag agagctcttg
     1561 cgacagcagg tcaaggagat caccttcctg aagaatacgg tgatggaatg tgacgcctgc
     1621 ggtctagaca gaaacctgcc tccactagca cctctgggcc cacacccaga aacgctggtg
     1681 aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg gttacatcga actggatctc
     1741 aacagcggta agatccttga gagttttcgc cccgaagaac gttttccaat gatgagcact
     1801 tttaaagttc tgctatgtgg cgcggtatta tcccgtattg acgccgggca agagcaactc
     1861 ggtcgccgca tacactattc tcagaatgac ttggttgagt actcaccagt cacagaaaag
     1921 catcttacgg atggcatgac agtaagagaa ttatgcagtg ctgccataac catgagtgat
     1981 aacactgcgg ccaacttact tctgacaacg atcggaggac cgaaggagct aaccgctttt
     2041 ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa
     2101 gccataccaa acgacgagcg tgacaccacg atgcctgtag caatggcaac aacgttgcgc
     2161 aaactattaa ctggcgaact acttactcta gcttcccggc aacaattaat agactggatg
     2221 gaggcggata aagttgcagg accacttctg cgctcggccc ttccggctgg ctggtttatt
     2281 gctgataaat ctggagccgg tgagcgtggg tctcgcggta tcattgcagc actggggcca
     2341 gatggtaagc cctcccgtat cgtagttatc tacacgacgg ggagtcaggc aactatggat
     2401 gaacgaaata gacagatcgc tgagataggt gcctcactga ttaagcattg gggccctgac
     2461 tacaaagacc atgacggtga ttataaagat catgacatcg attacaagga tgacgatgac
     2521 aagggccctc accaccacca tcaccactaa taataaggat cccccgacct cgacctctgg
     2581 ctaataaagg aaatttattt tcattgcaat agtgtgttgg aattttttgt gtctctcact
     2641 cggaaggaca tatgggaggg caaatcattt ggtcgagatc cctcggagat ctctagctag
     2701 aggatcgatc cccgccccgg acgaactaaa cctgactacg acatctctgc cccttcttcg
     2761 cggggcagtg catgtaatcc cttcagttgg ttggtacaac ttgccaactg ggccctgttc
     2821 cacatgtgac acgggggggg accaaacaca aaggggttct ctgactgtag ttgacatcct
     2881 tataaatgga tgtgcacatt tgccaacact gagtggcttt catcctggag cagactttgc
     2941 agtctgtgga ctgcaacaca acattgcctt tatgtgtaac tcttggctga agctcttaca
     3001 ccaatgctgg gggacatgta cctcccaggg gcccaggaag actacgggag gctacaccaa
     3061 cgtcaatcag aggggcctgt gtagctaccg ataagcggac cctcaagagg gcattagcaa
     3121 tagtgtttat aaggccccct tgttaaccct aaacgggtag catatgcttc ccgggtagta
     3181 gtatatacta tccagactaa ccctaattca atagcatatg ttacccaacg ggaagcatat
     3241 gctatcgaat tagggttagt aaaagggtcc taaggaacag cgatatctcc caccccatga
     3301 gctgtcacgg ttttatttac atggggtcag gattccacga gggtagtgaa ccattttagt
     3361 cacaagggca gtggctgaag atcaaggagc gggcagtgaa ctctcctgaa tcttcgcctg
     3421 cttcttcatt ctccttcgtt tagctaatag aataactgct gagttgtgaa cagtaaggtg
     3481 tatgtgaggt gctcgaaaac aaggtttcag gtgacgcccc cagaataaaa tttggacggg
     3541 gggttcagtg gtggcattgt gctatgacac caatataacc ctcacaaacc ccttgggcaa
     3601 taaatactag tgtaggaatg aaacattctg aatatcttta acaatagaaa tccatggggt
     3661 ggggacaagc cgtaaagact ggatgtccat ctcacacgaa tttatggcta tgggcaacac
     3721 ataatcctag tgcaatatga tactggggtt attaagatgt gtcccaggca gggaccaaga
     3781 caggtgaacc atgttgttac actctatttg taacaagggg aaagagagtg gacgccgaca
     3841 gcagcggact ccactggttg tctctaacac ccccgaaaat taaacggggc tccacgccaa
     3901 tggggcccat aaacaaagac aagtggccac tctttttttt gaaattgtgg agtgggggca
     3961 cgcgtcagcc cccacacgcc gccctgcggt tttggactgt aaaataaggg tgtaataact
     4021 tggctgattg taaccccgct aaccactgcg gtcaaaccac ttgcccacaa aaccactaat
     4081 ggcaccccgg ggaatacctg cataagtagg tgggcgggcc aagatagggg cgcgattgct
     4141 gcgatctgga ggacaaatta cacacacttg cgcctgagcg ccaagcacag ggttgttggt
     4201 cctcatattc acgaggtcgc tgagagcacg gtgggctaat gttgccatgg gtagcatata
     4261 ctacccaaat atctggatag catatgctat cctaatctat atctgggtag cataggctat
     4321 cctaatctat atctgggtag catatgctat cctaatctat atctgggtag tatatgctat
     4381 cctaatttat atctgggtag cataggctat cctaatctat atctgggtag catatgctat
     4441 cctaatctat atctgggtag tatatgctat cctaatctgt atccgggtag catatgctat
     4501 cctaatagag attagggtag tatatgctat cctaatttat atctgggtag catatactac
     4561 ccaaatatct ggatagcata tgctatccta atctatatct gggtagcata tgctatccta
     4621 atctatatct gggtagcata ggctatccta atctatatct gggtagcata tgctatccta
     4681 atctatatct gggtagtata tgctatccta atttatatct gggtagcata ggctatccta
     4741 atctatatct gggtagcata tgctatccta atctatatct gggtagtata tgctatccta
     4801 atctgtatcc gggtagcata tgctatcctc atgataagct gtcaaacatg agaattttct
     4861 tgaagacgaa agggcctcgt gatacgccta tttttatagg ttaatgtcat gataataatg
     4921 gtttcttaga cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta
     4981 tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt
     5041 caataatatt gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc ccttattccc
     5101 ttttttgcgg cattttgcct tcctgttttt gctcacccag aaacgctggt gaaagtaaaa
     5161 gatgctgaag atcagttggg tgcacgagtg ggttacatcg aactggatct caacagcggt
     5221 aagatccttg agagttttcg ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt
     5281 ctgctatgtg gcgcggtatt atcccgtgtt gacgccgggc aagagcaact cggtcgccgc
     5341 atacactatt ctcagaatga cttggttgag tactcaccag tcacagaaaa gcatcttacg
     5401 gatggcatga cagtaagaga attatgcagt gctgccataa ccatgagtga taacactgcg
     5461 gccaacttac ttctgacaac gatcggagga ccgaaggagc taaccgcttt tttgcacaac
     5521 atgggggatc atgtaactcg ccttgatcgt tgggaaccgg agctgaatga agccatacca
     5581 aacgacgagc gtgacaccac gatgcctgca gcaatggcaa caacgttgcg caaactatta
     5641 actggcgaac tacttactct agcttcccgg caacaattaa tagactggat ggaggcggat
     5701 aaagttgcag gaccacttct gcgctcggcc cttccggctg gctggtttat tgctgataaa
     5761 tctggagccg gtgagcgtgg gtctcgcggt atcattgcag cactggggcc agatggtaag
     5821 ccctcccgta tcgtagttat ctacacgacg gggagtcagg caactatgga tgaacgaaat
     5881 agacagatcg ctgagatagg tgcctcactg attaagcatt ggtaactgtc agaccaagtt
     5941 tactcatata tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg
     6001 aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga
     6061 gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta
     6121 atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa
     6181 gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact
     6241 gttcttctag tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca
     6301 tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt
     6361 accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg
     6421 ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag
     6481 cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta
     6541 agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat
     6601 ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg
     6661 tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc
     6721 ttttgctggc cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac
     6781 cgtattaccg cctttgagtg agctgatacc gctcgccgca gccgaacgac cgagcgcagc
     6841 gagtcagtga gcgaggaagc ggaagagcgc ccaatacgca aaccgcctct ccccgcgcgt
     6901 tggccgattc attaatgcag ctggcacgac aggtttcccg actggaaagc gggcagtgag
     6961 cgcaacgcaa ttaatgtgag ttagctcact cattaggcac cccaggcttt acactttatg
     7021 cttccggctc gtatgttgtg tggaattgtg agcggataac aatttcacac aggaaacagc
     7081 tatgaccatg attacgccaa gctctagcta gaggtcgacc aattctcatg tttgacagct
     7141 tatcatcgca gatccgggca acgttgttgc cattgctgca ggcgcagaac tggtaggtat
     7201 ggaagatcta tacattgaat caatattggc aattagccat attagtcatt ggttatatag
     7261 cataaatcaa tattggctat tggccattgc atacgttgta tctatatcat aatatgtaca
     7321 tttatattgg ctcatgtcca atatgaccgc catgttgaca ttgattattg actagttatt
     7381 aatagtaatc aattacgggg tcattagttc atagcccata tatggagttc cgcgttacat
     7441 aacttacggt aaatggcccg cctggctgac cgcccaacga cccccgccca ttgacgtcaa
     7501 taatgacgta tgttcccata gtaacgccaa tagggacttt ccattgacgt caatgggtgg
     7561 agtatttacg gtaaactgcc cacttggcag tacatcaagt gtatcatatg ccaagtccgc
     7621 cccctattga cgtcaatgac ggtaaatggc ccgcctggca ttatgcccag tacatgacct
     7681 tacgggactt tcctacttgg cagtacatct acgtattagt catcgctatt accatggtga
     7741 tgcggttttg gcagtacacc aatgggcgtg gatagcggtt tgactcacgg ggatttccaa
     7801 gtctccaccc cattgacgtc aatgggagtt tgttttggca ccaaaatcaa cgggactttc
     7861 caaaatgtcg taataacccc gccccgttga cgcaaatggg cggtaggcgt gtacggtggg
     7921 aggtctatat aagcagagct cgtttagtga accgtcagat cctcactctc ttccgcatcg
     7981 ctgtctgcga gggccagctg ttgggctcgc ggttgaggac aaactcttcg cggtctttcc
     8041 agtactcttg gatcggaaac ccgtcggcct ccgaacggta ctccgccacc gagggacctg
     8101 agcgagtccg catcgaccgg atcggaaaac ct
//
