LOCUS       sequence-224476-        8117 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 8117)
  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 8117)
  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_0526900-COMP-blac-flag-his
FEATURES             Location/Qualifiers
     source          1..8117
                     /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             1651..2436
                     /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             2443..2508
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             2515..2532
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    2568..2623
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(2622..2641)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(4859..4877)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4945..5049
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5050..5118,5119..5910)
                     /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      6081..6669
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6570..6589
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6823..6840
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    6957..6978
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6993..6998,6999..7016,7017..7023)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7031..7047
                     /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     7036..7058
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7055..7071
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7055..7071
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     enhancer        7342..7721
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        7722..7925
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     7876..7896
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     7922..7946
                     /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 ggccgccgcc
      421 tacttcttcg tgaaagaggg cgtggacaag tgcttcgtgg aaagcgtggc caacaacgtg
      481 gtcatcaccg ccacctacga caactacggc ctgaaggacg tgaagtgcct gatcaacatc
      541 aaggaccagc agggcaaggt gctgtactcc cacgacacca gcaagatccg gaagggcaag
      601 atcagctacc tgagcaccaa ggacggcctg taccacatct gcatcagctg ccccagcacc
      661 aactggttca agagcaccgc catcaagtgg tccctgagca tcgaagtggg cggcagcgac
      721 atcgaccccg agaacctggc caagaagtcc gagctgagcg agacactgac catcctgaac
      781 aacctgaaga aaaagttcaa cagcatgaag ctgcagcaga tctaccagaa acagatggcc
      841 agcaacctgt acgagtacaa caaggccgtg cacaacggcg cgccgtcgac ctccatcacg
      901 gcctataaga gtgaggggga gtcagcggag ttctccttcc cactcaacct tggagaggaa
      961 agcctgcagg gagagttgag atggaaggca gagaaggctc cttcttccca gtcctggatc
     1021 accttctccc taaagaacca aaaggtgtct gtgcagaagt ctactagcaa ccccaagttc
     1081 cagctgtccg aaacgctccc actcaccctt cagatacccc aggtctccct tcagtttgct
     1141 ggttctggca acctgaccct gactctggac agagggatac tgtatcagga agtgaacctg
     1201 gtggtgatga aagtgactca gcccgacagc aacactttga cctgtgaggt gatgggaccc
     1261 acctcaccca agatgagact gatcttgaag caggagaatc aggaggccag ggtctccagg
     1321 caggagaaag tgattcaagt gcaggcccct gaagcagggg tgtggcaatg tctactgagt
     1381 gaaggtgaag aggtcaagat ggactccaag atccaggttt tatccaaagg gttgaattcc
     1441 ggcggcggct ccggaggcgg taccggtggc ggcgacctag ccccacagat gcttcgagaa
     1501 ctccaggaga ctaatgcggc gctgcaagac gtgagagagc tcttgcgaca gcaggtcaag
     1561 gagatcacct tcctgaagaa tacggtgatg gaatgtgacg cctgcggtct agacagaaac
     1621 ctgcctccac tagcacctct gggcccacac ccagaaacgc tggtgaaagt aaaagatgct
     1681 gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag cggtaagatc
     1741 cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa agttctgcta
     1801 tgtggcgcgg tattatcccg tattgacgcc gggcaagagc aactcggtcg ccgcatacac
     1861 tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct tacggatggc
     1921 atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac
     1981 ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca caacatgggg
     2041 gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat accaaacgac
     2101 gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact attaactggc
     2161 gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc ggataaagtt
     2221 gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga taaatctgga
     2281 gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg taagccctcc
     2341 cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag
     2401 atcgctgaga taggtgcctc actgattaag cattggggcc ctgactacaa agaccatgac
     2461 ggtgattata aagatcatga catcgattac aaggatgacg atgacaaggg ccctcaccac
     2521 caccatcacc actaataata aggatccccc gacctcgacc tctggctaat aaaggaaatt
     2581 tattttcatt gcaatagtgt gttggaattt tttgtgtctc tcactcggaa ggacatatgg
     2641 gagggcaaat catttggtcg agatccctcg gagatctcta gctagaggat cgatccccgc
     2701 cccggacgaa ctaaacctga ctacgacatc tctgcccctt cttcgcgggg cagtgcatgt
     2761 aatcccttca gttggttggt acaacttgcc aactgggccc tgttccacat gtgacacggg
     2821 gggggaccaa acacaaaggg gttctctgac tgtagttgac atccttataa atggatgtgc
     2881 acatttgcca acactgagtg gctttcatcc tggagcagac tttgcagtct gtggactgca
     2941 acacaacatt gcctttatgt gtaactcttg gctgaagctc ttacaccaat gctgggggac
     3001 atgtacctcc caggggccca ggaagactac gggaggctac accaacgtca atcagagggg
     3061 cctgtgtagc taccgataag cggaccctca agagggcatt agcaatagtg tttataaggc
     3121 ccccttgtta accctaaacg ggtagcatat gcttcccggg tagtagtata tactatccag
     3181 actaacccta attcaatagc atatgttacc caacgggaag catatgctat cgaattaggg
     3241 ttagtaaaag ggtcctaagg aacagcgata tctcccaccc catgagctgt cacggtttta
     3301 tttacatggg gtcaggattc cacgagggta gtgaaccatt ttagtcacaa gggcagtggc
     3361 tgaagatcaa ggagcgggca gtgaactctc ctgaatcttc gcctgcttct tcattctcct
     3421 tcgtttagct aatagaataa ctgctgagtt gtgaacagta aggtgtatgt gaggtgctcg
     3481 aaaacaaggt ttcaggtgac gcccccagaa taaaatttgg acggggggtt cagtggtggc
     3541 attgtgctat gacaccaata taaccctcac aaaccccttg ggcaataaat actagtgtag
     3601 gaatgaaaca ttctgaatat ctttaacaat agaaatccat ggggtgggga caagccgtaa
     3661 agactggatg tccatctcac acgaatttat ggctatgggc aacacataat cctagtgcaa
     3721 tatgatactg gggttattaa gatgtgtccc aggcagggac caagacaggt gaaccatgtt
     3781 gttacactct atttgtaaca aggggaaaga gagtggacgc cgacagcagc ggactccact
     3841 ggttgtctct aacacccccg aaaattaaac ggggctccac gccaatgggg cccataaaca
     3901 aagacaagtg gccactcttt tttttgaaat tgtggagtgg gggcacgcgt cagcccccac
     3961 acgccgccct gcggttttgg actgtaaaat aagggtgtaa taacttggct gattgtaacc
     4021 ccgctaacca ctgcggtcaa accacttgcc cacaaaacca ctaatggcac cccggggaat
     4081 acctgcataa gtaggtgggc gggccaagat aggggcgcga ttgctgcgat ctggaggaca
     4141 aattacacac acttgcgcct gagcgccaag cacagggttg ttggtcctca tattcacgag
     4201 gtcgctgaga gcacggtggg ctaatgttgc catgggtagc atatactacc caaatatctg
     4261 gatagcatat gctatcctaa tctatatctg ggtagcatag gctatcctaa tctatatctg
     4321 ggtagcatat gctatcctaa tctatatctg ggtagtatat gctatcctaa tttatatctg
     4381 ggtagcatag gctatcctaa tctatatctg ggtagcatat gctatcctaa tctatatctg
     4441 ggtagtatat gctatcctaa tctgtatccg ggtagcatat gctatcctaa tagagattag
     4501 ggtagtatat gctatcctaa tttatatctg ggtagcatat actacccaaa tatctggata
     4561 gcatatgcta tcctaatcta tatctgggta gcatatgcta tcctaatcta tatctgggta
     4621 gcataggcta tcctaatcta tatctgggta gcatatgcta tcctaatcta tatctgggta
     4681 gtatatgcta tcctaattta tatctgggta gcataggcta tcctaatcta tatctgggta
     4741 gcatatgcta tcctaatcta tatctgggta gtatatgcta tcctaatctg tatccgggta
     4801 gcatatgcta tcctcatgat aagctgtcaa acatgagaat tttcttgaag acgaaagggc
     4861 ctcgtgatac gcctattttt ataggttaat gtcatgataa taatggtttc ttagacgtca
     4921 ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat
     4981 tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa
     5041 aggaagagta tgagtattca acatttccgt gtcgccctta ttcccttttt tgcggcattt
     5101 tgccttcctg tttttgctca cccagaaacg ctggtgaaag taaaagatgc tgaagatcag
     5161 ttgggtgcac gagtgggtta catcgaactg gatctcaaca gcggtaagat ccttgagagt
     5221 tttcgccccg aagaacgttt tccaatgatg agcactttta aagttctgct atgtggcgcg
     5281 gtattatccc gtgttgacgc cgggcaagag caactcggtc gccgcataca ctattctcag
     5341 aatgacttgg ttgagtactc accagtcaca gaaaagcatc ttacggatgg catgacagta
     5401 agagaattat gcagtgctgc cataaccatg agtgataaca ctgcggccaa cttacttctg
     5461 acaacgatcg gaggaccgaa ggagctaacc gcttttttgc acaacatggg ggatcatgta
     5521 actcgccttg atcgttggga accggagctg aatgaagcca taccaaacga cgagcgtgac
     5581 accacgatgc ctgcagcaat ggcaacaacg ttgcgcaaac tattaactgg cgaactactt
     5641 actctagctt cccggcaaca attaatagac tggatggagg cggataaagt tgcaggacca
     5701 cttctgcgct cggcccttcc ggctggctgg tttattgctg ataaatctgg agccggtgag
     5761 cgtgggtctc gcggtatcat tgcagcactg gggccagatg gtaagccctc ccgtatcgta
     5821 gttatctaca cgacggggag tcaggcaact atggatgaac gaaatagaca gatcgctgag
     5881 ataggtgcct cactgattaa gcattggtaa ctgtcagacc aagtttactc atatatactt
     5941 tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat
     6001 aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta
     6061 gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa
     6121 acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt
     6181 tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct tctagtgtag
     6241 ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta
     6301 atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca
     6361 agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag
     6421 cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa
     6481 agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga
     6541 acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc
     6601 gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc
     6661 ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt
     6721 gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat taccgccttt
     6781 gagtgagctg ataccgctcg ccgcagccga acgaccgagc gcagcgagtc agtgagcgag
     6841 gaagcggaag agcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa
     6901 tgcagctggc acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat
     6961 gtgagttagc tcactcatta ggcaccccag gctttacact ttatgcttcc ggctcgtatg
     7021 ttgtgtggaa ttgtgagcgg ataacaattt cacacaggaa acagctatga ccatgattac
     7081 gccaagctct agctagaggt cgaccaattc tcatgtttga cagcttatca tcgcagatcc
     7141 gggcaacgtt gttgccattg ctgcaggcgc agaactggta ggtatggaag atctatacat
     7201 tgaatcaata ttggcaatta gccatattag tcattggtta tatagcataa atcaatattg
     7261 gctattggcc attgcatacg ttgtatctat atcataatat gtacatttat attggctcat
     7321 gtccaatatg accgccatgt tgacattgat tattgactag ttattaatag taatcaatta
     7381 cggggtcatt agttcatagc ccatatatgg agttccgcgt tacataactt acggtaaatg
     7441 gcccgcctgg ctgaccgccc aacgaccccc gcccattgac gtcaataatg acgtatgttc
     7501 ccatagtaac gccaataggg actttccatt gacgtcaatg ggtggagtat ttacggtaaa
     7561 ctgcccactt ggcagtacat caagtgtatc atatgccaag tccgccccct attgacgtca
     7621 atgacggtaa atggcccgcc tggcattatg cccagtacat gaccttacgg gactttccta
     7681 cttggcagta catctacgta ttagtcatcg ctattaccat ggtgatgcgg ttttggcagt
     7741 acaccaatgg gcgtggatag cggtttgact cacggggatt tccaagtctc caccccattg
     7801 acgtcaatgg gagtttgttt tggcaccaaa atcaacggga ctttccaaaa tgtcgtaata
     7861 accccgcccc gttgacgcaa atgggcggta ggcgtgtacg gtgggaggtc tatataagca
     7921 gagctcgttt agtgaaccgt cagatcctca ctctcttccg catcgctgtc tgcgagggcc
     7981 agctgttggg ctcgcggttg aggacaaact cttcgcggtc tttccagtac tcttggatcg
     8041 gaaacccgtc ggcctccgaa cggtactccg ccaccgaggg acctgagcga gtccgcatcg
     8101 accggatcgg aaaacct
//
