LOCUS       sequence-224245-        8183 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 8183)
  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 8183)
  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_1350600-COMP-blac-flag-his
FEATURES             Location/Qualifiers
     source          1..8183
                     /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             1717..2502
                     /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             2509..2574
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             2581..2598
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    2634..2689
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(2688..2707)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(4925..4943)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5011..5115
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5116..5184,5185..5976)
                     /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      6147..6735
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6636..6655
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6889..6906
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    7023..7044
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7059..7064,7065..7082,7083..7089)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7097..7113
                     /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     7102..7124
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7121..7137
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7121..7137
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     enhancer        7408..7787
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        7788..7991
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     7942..7962
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     7988..8012
                     /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 ggccgccatg
      421 aacgtggtca tccccagaaa caaagagtcc cggaccaaga gagtgatcaa gggcatcaag
      481 aagaagtaca acggcttcac cgagcgctac agaaagagct tccacctgaa ggacgaggac
      541 gtgatcggcg catacagcat gcaggcctac tactttttcg acaagaacat cgagcacatg
      601 ttcaagtggc ggctgttcta caacttctgc aagaagtccc ctagcccctg caactacttc
      661 aagctgcgga aagagttcaa gcggatctac gagggcagca ccttcaccct gaagaaatac
      721 gtgtccgaag tgaacatcat cttcaaccag ttcatcaaag agtacagcac cgtccggttc
      781 aacgtggacc tgaagcagaa ccacattgcc aacctggcct acacccctaa cctggccacc
      841 cagatcatga agtccctgac cagcctgatc aacgaccagc tgatggaaaa cgagctgaag
      901 aaccccgcct ggaagctgag cttcaactac ctgaccggca ccggcgcgcc gtcgacctcc
      961 atcacggcct ataagagtga gggggagtca gcggagttct ccttcccact caaccttgga
     1021 gaggaaagcc tgcagggaga gttgagatgg aaggcagaga aggctccttc ttcccagtcc
     1081 tggatcacct tctccctaaa gaaccaaaag gtgtctgtgc agaagtctac tagcaacccc
     1141 aagttccagc tgtccgaaac gctcccactc acccttcaga taccccaggt ctcccttcag
     1201 tttgctggtt ctggcaacct gaccctgact ctggacagag ggatactgta tcaggaagtg
     1261 aacctggtgg tgatgaaagt gactcagccc gacagcaaca ctttgacctg tgaggtgatg
     1321 ggacccacct cacccaagat gagactgatc ttgaagcagg agaatcagga ggccagggtc
     1381 tccaggcagg agaaagtgat tcaagtgcag gcccctgaag caggggtgtg gcaatgtcta
     1441 ctgagtgaag gtgaagaggt caagatggac tccaagatcc aggttttatc caaagggttg
     1501 aattccggcg gcggctccgg aggcggtacc ggtggcggcg acctagcccc acagatgctt
     1561 cgagaactcc aggagactaa tgcggcgctg caagacgtga gagagctctt gcgacagcag
     1621 gtcaaggaga tcaccttcct gaagaatacg gtgatggaat gtgacgcctg cggtctagac
     1681 agaaacctgc ctccactagc acctctgggc ccacacccag aaacgctggt gaaagtaaaa
     1741 gatgctgaag atcagttggg tgcacgagtg ggttacatcg aactggatct caacagcggt
     1801 aagatccttg agagttttcg ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt
     1861 ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc aagagcaact cggtcgccgc
     1921 atacactatt ctcagaatga cttggttgag tactcaccag tcacagaaaa gcatcttacg
     1981 gatggcatga cagtaagaga attatgcagt gctgccataa ccatgagtga taacactgcg
     2041 gccaacttac ttctgacaac gatcggagga ccgaaggagc taaccgcttt tttgcacaac
     2101 atgggggatc atgtaactcg ccttgatcgt tgggaaccgg agctgaatga agccatacca
     2161 aacgacgagc gtgacaccac gatgcctgta gcaatggcaa caacgttgcg caaactatta
     2221 actggcgaac tacttactct agcttcccgg caacaattaa tagactggat ggaggcggat
     2281 aaagttgcag gaccacttct gcgctcggcc cttccggctg gctggtttat tgctgataaa
     2341 tctggagccg gtgagcgtgg gtctcgcggt atcattgcag cactggggcc agatggtaag
     2401 ccctcccgta tcgtagttat ctacacgacg gggagtcagg caactatgga tgaacgaaat
     2461 agacagatcg ctgagatagg tgcctcactg attaagcatt ggggccctga ctacaaagac
     2521 catgacggtg attataaaga tcatgacatc gattacaagg atgacgatga caagggccct
     2581 caccaccacc atcaccacta ataataagga tcccccgacc tcgacctctg gctaataaag
     2641 gaaatttatt ttcattgcaa tagtgtgttg gaattttttg tgtctctcac tcggaaggac
     2701 atatgggagg gcaaatcatt tggtcgagat ccctcggaga tctctagcta gaggatcgat
     2761 ccccgccccg gacgaactaa acctgactac gacatctctg ccccttcttc gcggggcagt
     2821 gcatgtaatc ccttcagttg gttggtacaa cttgccaact gggccctgtt ccacatgtga
     2881 cacggggggg gaccaaacac aaaggggttc tctgactgta gttgacatcc ttataaatgg
     2941 atgtgcacat ttgccaacac tgagtggctt tcatcctgga gcagactttg cagtctgtgg
     3001 actgcaacac aacattgcct ttatgtgtaa ctcttggctg aagctcttac accaatgctg
     3061 ggggacatgt acctcccagg ggcccaggaa gactacggga ggctacacca acgtcaatca
     3121 gaggggcctg tgtagctacc gataagcgga ccctcaagag ggcattagca atagtgttta
     3181 taaggccccc ttgttaaccc taaacgggta gcatatgctt cccgggtagt agtatatact
     3241 atccagacta accctaattc aatagcatat gttacccaac gggaagcata tgctatcgaa
     3301 ttagggttag taaaagggtc ctaaggaaca gcgatatctc ccaccccatg agctgtcacg
     3361 gttttattta catggggtca ggattccacg agggtagtga accattttag tcacaagggc
     3421 agtggctgaa gatcaaggag cgggcagtga actctcctga atcttcgcct gcttcttcat
     3481 tctccttcgt ttagctaata gaataactgc tgagttgtga acagtaaggt gtatgtgagg
     3541 tgctcgaaaa caaggtttca ggtgacgccc ccagaataaa atttggacgg ggggttcagt
     3601 ggtggcattg tgctatgaca ccaatataac cctcacaaac cccttgggca ataaatacta
     3661 gtgtaggaat gaaacattct gaatatcttt aacaatagaa atccatgggg tggggacaag
     3721 ccgtaaagac tggatgtcca tctcacacga atttatggct atgggcaaca cataatccta
     3781 gtgcaatatg atactggggt tattaagatg tgtcccaggc agggaccaag acaggtgaac
     3841 catgttgtta cactctattt gtaacaaggg gaaagagagt ggacgccgac agcagcggac
     3901 tccactggtt gtctctaaca cccccgaaaa ttaaacgggg ctccacgcca atggggccca
     3961 taaacaaaga caagtggcca ctcttttttt tgaaattgtg gagtgggggc acgcgtcagc
     4021 ccccacacgc cgccctgcgg ttttggactg taaaataagg gtgtaataac ttggctgatt
     4081 gtaaccccgc taaccactgc ggtcaaacca cttgcccaca aaaccactaa tggcaccccg
     4141 gggaatacct gcataagtag gtgggcgggc caagataggg gcgcgattgc tgcgatctgg
     4201 aggacaaatt acacacactt gcgcctgagc gccaagcaca gggttgttgg tcctcatatt
     4261 cacgaggtcg ctgagagcac ggtgggctaa tgttgccatg ggtagcatat actacccaaa
     4321 tatctggata gcatatgcta tcctaatcta tatctgggta gcataggcta tcctaatcta
     4381 tatctgggta gcatatgcta tcctaatcta tatctgggta gtatatgcta tcctaattta
     4441 tatctgggta gcataggcta tcctaatcta tatctgggta gcatatgcta tcctaatcta
     4501 tatctgggta gtatatgcta tcctaatctg tatccgggta gcatatgcta tcctaataga
     4561 gattagggta gtatatgcta tcctaattta tatctgggta gcatatacta cccaaatatc
     4621 tggatagcat atgctatcct aatctatatc tgggtagcat atgctatcct aatctatatc
     4681 tgggtagcat aggctatcct aatctatatc tgggtagcat atgctatcct aatctatatc
     4741 tgggtagtat atgctatcct aatttatatc tgggtagcat aggctatcct aatctatatc
     4801 tgggtagcat atgctatcct aatctatatc tgggtagtat atgctatcct aatctgtatc
     4861 cgggtagcat atgctatcct catgataagc tgtcaaacat gagaattttc ttgaagacga
     4921 aagggcctcg tgatacgcct atttttatag gttaatgtca tgataataat ggtttcttag
     4981 acgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa
     5041 atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat
     5101 tgaaaaagga agagtatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg
     5161 gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa
     5221 gatcagttgg gtgcacgagt gggttacatc gaactggatc tcaacagcgg taagatcctt
     5281 gagagttttc gccccgaaga acgttttcca atgatgagca cttttaaagt tctgctatgt
     5341 ggcgcggtat tatcccgtgt tgacgccggg caagagcaac tcggtcgccg catacactat
     5401 tctcagaatg acttggttga gtactcacca gtcacagaaa agcatcttac ggatggcatg
     5461 acagtaagag aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta
     5521 cttctgacaa cgatcggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat
     5581 catgtaactc gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag
     5641 cgtgacacca cgatgcctgc agcaatggca acaacgttgc gcaaactatt aactggcgaa
     5701 ctacttactc tagcttcccg gcaacaatta atagactgga tggaggcgga taaagttgca
     5761 ggaccacttc tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc
     5821 ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt
     5881 atcgtagtta tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc
     5941 gctgagatag gtgcctcact gattaagcat tggtaactgt cagaccaagt ttactcatat
     6001 atactttaga ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt
     6061 tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac
     6121 cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc
     6181 ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca
     6241 actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta
     6301 gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct
     6361 ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg
     6421 gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc
     6481 acacagccca gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta
     6541 tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg
     6601 gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt
     6661 cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg
     6721 cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg
     6781 ccttttgctc acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc
     6841 gcctttgagt gagctgatac cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg
     6901 agcgaggaag cggaagagcg cccaatacgc aaaccgcctc tccccgcgcg ttggccgatt
     6961 cattaatgca gctggcacga caggtttccc gactggaaag cgggcagtga gcgcaacgca
     7021 attaatgtga gttagctcac tcattaggca ccccaggctt tacactttat gcttccggct
     7081 cgtatgttgt gtggaattgt gagcggataa caatttcaca caggaaacag ctatgaccat
     7141 gattacgcca agctctagct agaggtcgac caattctcat gtttgacagc ttatcatcgc
     7201 agatccgggc aacgttgttg ccattgctgc aggcgcagaa ctggtaggta tggaagatct
     7261 atacattgaa tcaatattgg caattagcca tattagtcat tggttatata gcataaatca
     7321 atattggcta ttggccattg catacgttgt atctatatca taatatgtac atttatattg
     7381 gctcatgtcc aatatgaccg ccatgttgac attgattatt gactagttat taatagtaat
     7441 caattacggg gtcattagtt catagcccat atatggagtt ccgcgttaca taacttacgg
     7501 taaatggccc gcctggctga ccgcccaacg acccccgccc attgacgtca ataatgacgt
     7561 atgttcccat agtaacgcca atagggactt tccattgacg tcaatgggtg gagtatttac
     7621 ggtaaactgc ccacttggca gtacatcaag tgtatcatat gccaagtccg ccccctattg
     7681 acgtcaatga cggtaaatgg cccgcctggc attatgccca gtacatgacc ttacgggact
     7741 ttcctacttg gcagtacatc tacgtattag tcatcgctat taccatggtg atgcggtttt
     7801 ggcagtacac caatgggcgt ggatagcggt ttgactcacg gggatttcca agtctccacc
     7861 ccattgacgt caatgggagt ttgttttggc accaaaatca acgggacttt ccaaaatgtc
     7921 gtaataaccc cgccccgttg acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata
     7981 taagcagagc tcgtttagtg aaccgtcaga tcctcactct cttccgcatc gctgtctgcg
     8041 agggccagct gttgggctcg cggttgagga caaactcttc gcggtctttc cagtactctt
     8101 ggatcggaaa cccgtcggcc tccgaacggt actccgccac cgagggacct gagcgagtcc
     8161 gcatcgaccg gatcggaaaa cct
//
