LOCUS       sequence-208744-        8173 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 8173)
  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 8173)
  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: P25-COMP-blac-flag-his
FEATURES             Location/Qualifiers
     source          1..8173
                     /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             1705..2490
                     /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             2497..2562
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             2569..2586
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    2624..2679
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(2678..2697)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(4915..4933)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5001..5105
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5106..5174,5175..5966)
                     /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      6137..6725
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6626..6645
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6879..6896
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    7013..7034
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7049..7054,7055..7072,7073..7079)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7087..7103
                     /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     7092..7114
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7111..7127
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7111..7127
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     enhancer        7398..7777
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        7778..7981
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     7932..7952
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     7978..8002
                     /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 ggccgccaaa
      421 gtgaccgtgg acaccgtgtg caagcggggc ttcctgatcc agatgagcgg ccacctggaa
      481 tgcaagtgcg agaacgacct ggtgctggtc aacgaggaaa cctgcgaaga gaaggtgctg
      541 aagtgcgacg agaaaaccgt gaacaagccc tgcggcgact tcagcaagtg catcaagatc
      601 gacggcaacc ccgtgtccta cgcctgcaag tgcaacctgg gctacgacat ggtcaacaac
      661 gtgtgcatcc ccaacgagtg caagaacgtg gcctgcggca acggcaagtg tatcctggac
      721 accagcaacc ctgtgaaaac cggcgtgtgc tcctgcaaca tcggcaaggt gcccaacgtg
      781 caggaccaga acaagtgcag caaggacggc gagacaaagt gcagcctgaa gtgcctgaaa
      841 gagaatgagg cctgcaaggc cgtggacggc atctacaagt gcgactgcaa ggatggcttc
      901 atcatcgaca acgaggccag catctgcacc ggcgcgccgt cgacctccat cacggcctat
      961 aagagtgagg gggagtcagc ggagttctcc ttcccactca accttggaga ggaaagcctg
     1021 cagggagagt tgagatggaa ggcagagaag gctccttctt cccagtcctg gatcaccttc
     1081 tccctaaaga accaaaaggt gtctgtgcag aagtctacta gcaaccccaa gttccagctg
     1141 tccgaaacgc tcccactcac ccttcagata ccccaggtct cccttcagtt tgctggttct
     1201 ggcaacctga ccctgactct ggacagaggg atactgtatc aggaagtgaa cctggtggtg
     1261 atgaaagtga ctcagcccga cagcaacact ttgacctgtg aggtgatggg acccacctca
     1321 cccaagatga gactgatctt gaagcaggag aatcaggagg ccagggtctc caggcaggag
     1381 aaagtgattc aagtgcaggc ccctgaagca ggggtgtggc aatgtctact gagtgaaggt
     1441 gaagaggtca agatggactc caagatccag gttttatcca aagggttgaa ttccggcggc
     1501 ggctccggag gcggtaccgg tggcggcgac ctagccccac agatgcttcg agaactccag
     1561 gagactaatg cggcgctgca agacgtgaga gagctcttgc gacagcaggt caaggagatc
     1621 accttcctga agaatacggt gatggaatgt gacgcctgcg gtctagacag aaacctgcct
     1681 ccactagcac ctctgggccc acacccagaa acgctggtga aagtaaaaga tgctgaagat
     1741 cagttgggtg cacgagtggg ttacatcgaa ctggatctca acagcggtaa gatccttgag
     1801 agttttcgcc ccgaagaacg ttttccaatg atgagcactt ttaaagttct gctatgtggc
     1861 gcggtattat cccgtattga cgccgggcaa gagcaactcg gtcgccgcat acactattct
     1921 cagaatgact tggttgagta ctcaccagtc acagaaaagc atcttacgga tggcatgaca
     1981 gtaagagaat tatgcagtgc tgccataacc atgagtgata acactgcggc caacttactt
     2041 ctgacaacga tcggaggacc gaaggagcta accgcttttt tgcacaacat gggggatcat
     2101 gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt
     2161 gacaccacga tgcctgtagc aatggcaaca acgttgcgca aactattaac tggcgaacta
     2221 cttactctag cttcccggca acaattaata gactggatgg aggcggataa agttgcagga
     2281 ccacttctgc gctcggccct tccggctggc tggtttattg ctgataaatc tggagccggt
     2341 gagcgtgggt ctcgcggtat cattgcagca ctggggccag atggtaagcc ctcccgtatc
     2401 gtagttatct acacgacggg gagtcaggca actatggatg aacgaaatag acagatcgct
     2461 gagataggtg cctcactgat taagcattgg ggccctgact acaaagacca tgacggtgat
     2521 tataaagatc atgacatcga ttacaaggat gacgatgaca agggccctca ccaccaccat
     2581 caccactaat taattaagga tcccccgacc tcgacctctg gctaataaag gaaatttatt
     2641 ttcattgcaa tagtgtgttg gaattttttg tgtctctcac tcggaaggac atatgggagg
     2701 gcaaatcatt tggtcgagat ccctcggaga tctctagcta gaggatcgat ccccgccccg
     2761 gacgaactaa acctgactac gacatctctg ccccttcttc gcggggcagt gcatgtaatc
     2821 ccttcagttg gttggtacaa cttgccaact gggccctgtt ccacatgtga cacggggggg
     2881 gaccaaacac aaaggggttc tctgactgta gttgacatcc ttataaatgg atgtgcacat
     2941 ttgccaacac tgagtggctt tcatcctgga gcagactttg cagtctgtgg actgcaacac
     3001 aacattgcct ttatgtgtaa ctcttggctg aagctcttac accaatgctg ggggacatgt
     3061 acctcccagg ggcccaggaa gactacggga ggctacacca acgtcaatca gaggggcctg
     3121 tgtagctacc gataagcgga ccctcaagag ggcattagca atagtgttta taaggccccc
     3181 ttgttaaccc taaacgggta gcatatgctt cccgggtagt agtatatact atccagacta
     3241 accctaattc aatagcatat gttacccaac gggaagcata tgctatcgaa ttagggttag
     3301 taaaagggtc ctaaggaaca gcgatatctc ccaccccatg agctgtcacg gttttattta
     3361 catggggtca ggattccacg agggtagtga accattttag tcacaagggc agtggctgaa
     3421 gatcaaggag cgggcagtga actctcctga atcttcgcct gcttcttcat tctccttcgt
     3481 ttagctaata gaataactgc tgagttgtga acagtaaggt gtatgtgagg tgctcgaaaa
     3541 caaggtttca ggtgacgccc ccagaataaa atttggacgg ggggttcagt ggtggcattg
     3601 tgctatgaca ccaatataac cctcacaaac cccttgggca ataaatacta gtgtaggaat
     3661 gaaacattct gaatatcttt aacaatagaa atccatgggg tggggacaag ccgtaaagac
     3721 tggatgtcca tctcacacga atttatggct atgggcaaca cataatccta gtgcaatatg
     3781 atactggggt tattaagatg tgtcccaggc agggaccaag acaggtgaac catgttgtta
     3841 cactctattt gtaacaaggg gaaagagagt ggacgccgac agcagcggac tccactggtt
     3901 gtctctaaca cccccgaaaa ttaaacgggg ctccacgcca atggggccca taaacaaaga
     3961 caagtggcca ctcttttttt tgaaattgtg gagtgggggc acgcgtcagc ccccacacgc
     4021 cgccctgcgg ttttggactg taaaataagg gtgtaataac ttggctgatt gtaaccccgc
     4081 taaccactgc ggtcaaacca cttgcccaca aaaccactaa tggcaccccg gggaatacct
     4141 gcataagtag gtgggcgggc caagataggg gcgcgattgc tgcgatctgg aggacaaatt
     4201 acacacactt gcgcctgagc gccaagcaca gggttgttgg tcctcatatt cacgaggtcg
     4261 ctgagagcac ggtgggctaa tgttgccatg ggtagcatat actacccaaa tatctggata
     4321 gcatatgcta tcctaatcta tatctgggta gcataggcta tcctaatcta tatctgggta
     4381 gcatatgcta tcctaatcta tatctgggta gtatatgcta tcctaattta tatctgggta
     4441 gcataggcta tcctaatcta tatctgggta gcatatgcta tcctaatcta tatctgggta
     4501 gtatatgcta tcctaatctg tatccgggta gcatatgcta tcctaataga gattagggta
     4561 gtatatgcta tcctaattta tatctgggta gcatatacta cccaaatatc tggatagcat
     4621 atgctatcct aatctatatc tgggtagcat atgctatcct aatctatatc tgggtagcat
     4681 aggctatcct aatctatatc tgggtagcat atgctatcct aatctatatc tgggtagtat
     4741 atgctatcct aatttatatc tgggtagcat aggctatcct aatctatatc tgggtagcat
     4801 atgctatcct aatctatatc tgggtagtat atgctatcct aatctgtatc cgggtagcat
     4861 atgctatcct catgataagc tgtcaaacat gagaattttc ttgaagacga aagggcctcg
     4921 tgatacgcct atttttatag gttaatgtca tgataataat ggtttcttag acgtcaggtg
     4981 gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa
     5041 atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga
     5101 agagtatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc
     5161 ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg
     5221 gtgcacgagt gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc
     5281 gccccgaaga acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat
     5341 tatcccgtgt tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg
     5401 acttggttga gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag
     5461 aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa
     5521 cgatcggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc
     5581 gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca
     5641 cgatgcctgc agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc
     5701 tagcttcccg gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc
     5761 tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg
     5821 ggtctcgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta
     5881 tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag
     5941 gtgcctcact gattaagcat tggtaactgt cagaccaagt ttactcatat atactttaga
     6001 ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc
     6061 tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa
     6121 agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa
     6181 aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc
     6241 cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt
     6301 agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc
     6361 tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac
     6421 gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca
     6481 gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg
     6541 ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag
     6601 gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt
     6661 ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat
     6721 ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc
     6781 acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gcctttgagt
     6841 gagctgatac cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg agcgaggaag
     6901 cggaagagcg cccaatacgc aaaccgcctc tccccgcgcg ttggccgatt cattaatgca
     6961 gctggcacga caggtttccc gactggaaag cgggcagtga gcgcaacgca attaatgtga
     7021 gttagctcac tcattaggca ccccaggctt tacactttat gcttccggct cgtatgttgt
     7081 gtggaattgt gagcggataa caatttcaca caggaaacag ctatgaccat gattacgcca
     7141 agctctagct agaggtcgac caattctcat gtttgacagc ttatcatcgc agatccgggc
     7201 aacgttgttg ccattgctgc aggcgcagaa ctggtaggta tggaagatct atacattgaa
     7261 tcaatattgg caattagcca tattagtcat tggttatata gcataaatca atattggcta
     7321 ttggccattg catacgttgt atctatatca taatatgtac atttatattg gctcatgtcc
     7381 aatatgaccg ccatgttgac attgattatt gactagttat taatagtaat caattacggg
     7441 gtcattagtt catagcccat atatggagtt ccgcgttaca taacttacgg taaatggccc
     7501 gcctggctga ccgcccaacg acccccgccc attgacgtca ataatgacgt atgttcccat
     7561 agtaacgcca atagggactt tccattgacg tcaatgggtg gagtatttac ggtaaactgc
     7621 ccacttggca gtacatcaag tgtatcatat gccaagtccg ccccctattg acgtcaatga
     7681 cggtaaatgg cccgcctggc attatgccca gtacatgacc ttacgggact ttcctacttg
     7741 gcagtacatc tacgtattag tcatcgctat taccatggtg atgcggtttt ggcagtacac
     7801 caatgggcgt ggatagcggt ttgactcacg gggatttcca agtctccacc ccattgacgt
     7861 caatgggagt ttgttttggc accaaaatca acgggacttt ccaaaatgtc gtaataaccc
     7921 cgccccgttg acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata taagcagagc
     7981 tcgtttagtg aaccgtcaga tcctcactct cttccgcatc gctgtctgcg agggccagct
     8041 gttgggctcg cggttgagga caaactcttc gcggtctttc cagtactctt ggatcggaaa
     8101 cccgtcggcc tccgaacggt actccgccac cgagggacct gagcgagtcc gcatcgaccg
     8161 gatcggaaaa cct
//
