LOCUS       sequence-224279-        8387 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 8387)
  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 8387)
  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_1434400-COMP-blac-flag-his
FEATURES             Location/Qualifiers
     source          1..8387
                     /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             1921..2706
                     /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             2713..2778
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             2785..2802
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    2838..2893
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(2892..2911)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(5129..5147)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5215..5319
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5320..5388,5389..6180)
                     /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      6351..6939
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6840..6859
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7093..7110
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    7227..7248
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7263..7268,7269..7286,7287..7293)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7301..7317
                     /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     7306..7328
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7325..7341
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7325..7341
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     enhancer        7612..7991
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        7992..8195
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     8146..8166
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     8192..8216
                     /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 ggccgccaac
      421 aatatcgccc acaaagagga cgactgcggc gccagcgagc tggccatcaa gaagctgcag
      481 gaactgaaga agttcgagct ggacatcctg aaggacttta tcaagaagga caccgaccac
      541 accgacatgt acaagagata ccactgcatc agcagcgact ttctgaagaa caccaagcag
      601 gaagataaga aagaggacaa ctacgccaaa aaagaggata agaaggatat cgagaacagc
      661 gccaactaca tcatcaacaa caagaacgag aagatcagct ttatcaacaa gatcatcagc
      721 ttcgtgggca tgtggaacta caacaaatac ggcaacaaca tccaggaaag caagaacaag
      781 attaacatct acaacaacaa taacaacaag gccgagaacg agctgctgta caacaaagaa
      841 gattacacca accacatccc caccaacgag aaccccaaca gctacatgca gctgagcgcc
      901 ttcaacctgg ccaaaaagag catgttcctg ggcggcaagg acaacgccgc ccagaatttc
      961 gaagtgggca actacggcag cttctacatg atcttcggcg ccagaaacac cgactacccc
     1021 tgggcctgca gctgcgacac actgcagctg gaagagtaca aagagaaaaa gcggagctac
     1081 gtgctgtgca gcaaccagct ggatatgagc atccagaaca gcgacctgtt ctgcaacccc
     1141 aagaacggcg cgccgtcgac ctccatcacg gcctataaga gtgaggggga gtcagcggag
     1201 ttctccttcc cactcaacct tggagaggaa agcctgcagg gagagttgag atggaaggca
     1261 gagaaggctc cttcttccca gtcctggatc accttctccc taaagaacca aaaggtgtct
     1321 gtgcagaagt ctactagcaa ccccaagttc cagctgtccg aaacgctccc actcaccctt
     1381 cagatacccc aggtctccct tcagtttgct ggttctggca acctgaccct gactctggac
     1441 agagggatac tgtatcagga agtgaacctg gtggtgatga aagtgactca gcccgacagc
     1501 aacactttga cctgtgaggt gatgggaccc acctcaccca agatgagact gatcttgaag
     1561 caggagaatc aggaggccag ggtctccagg caggagaaag tgattcaagt gcaggcccct
     1621 gaagcagggg tgtggcaatg tctactgagt gaaggtgaag aggtcaagat ggactccaag
     1681 atccaggttt tatccaaagg gttgaattcc ggcggcggct ccggaggcgg taccggtggc
     1741 ggcgacctag ccccacagat gcttcgagaa ctccaggaga ctaatgcggc gctgcaagac
     1801 gtgagagagc tcttgcgaca gcaggtcaag gagatcacct tcctgaagaa tacggtgatg
     1861 gaatgtgacg cctgcggtct agacagaaac ctgcctccac tagcacctct gggcccacac
     1921 ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg agtgggttac
     1981 atcgaactgg atctcaacag cggtaagatc cttgagagtt ttcgccccga agaacgtttt
     2041 ccaatgatga gcacttttaa agttctgcta tgtggcgcgg tattatcccg tattgacgcc
     2101 gggcaagagc aactcggtcg ccgcatacac tattctcaga atgacttggt tgagtactca
     2161 ccagtcacag aaaagcatct tacggatggc atgacagtaa gagaattatg cagtgctgcc
     2221 ataaccatga gtgataacac tgcggccaac ttacttctga caacgatcgg aggaccgaag
     2281 gagctaaccg cttttttgca caacatgggg gatcatgtaa ctcgccttga tcgttgggaa
     2341 ccggagctga atgaagccat accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg
     2401 gcaacaacgt tgcgcaaact attaactggc gaactactta ctctagcttc ccggcaacaa
     2461 ttaatagact ggatggaggc ggataaagtt gcaggaccac ttctgcgctc ggcccttccg
     2521 gctggctggt ttattgctga taaatctgga gccggtgagc gtgggtctcg cggtatcatt
     2581 gcagcactgg ggccagatgg taagccctcc cgtatcgtag ttatctacac gacggggagt
     2641 caggcaacta tggatgaacg aaatagacag atcgctgaga taggtgcctc actgattaag
     2701 cattggggcc ctgactacaa agaccatgac ggtgattata aagatcatga catcgattac
     2761 aaggatgacg atgacaaggg ccctcaccac caccatcacc actaataata aggatccccc
     2821 gacctcgacc tctggctaat aaaggaaatt tattttcatt gcaatagtgt gttggaattt
     2881 tttgtgtctc tcactcggaa ggacatatgg gagggcaaat catttggtcg agatccctcg
     2941 gagatctcta gctagaggat cgatccccgc cccggacgaa ctaaacctga ctacgacatc
     3001 tctgcccctt cttcgcgggg cagtgcatgt aatcccttca gttggttggt acaacttgcc
     3061 aactgggccc tgttccacat gtgacacggg gggggaccaa acacaaaggg gttctctgac
     3121 tgtagttgac atccttataa atggatgtgc acatttgcca acactgagtg gctttcatcc
     3181 tggagcagac tttgcagtct gtggactgca acacaacatt gcctttatgt gtaactcttg
     3241 gctgaagctc ttacaccaat gctgggggac atgtacctcc caggggccca ggaagactac
     3301 gggaggctac accaacgtca atcagagggg cctgtgtagc taccgataag cggaccctca
     3361 agagggcatt agcaatagtg tttataaggc ccccttgtta accctaaacg ggtagcatat
     3421 gcttcccggg tagtagtata tactatccag actaacccta attcaatagc atatgttacc
     3481 caacgggaag catatgctat cgaattaggg ttagtaaaag ggtcctaagg aacagcgata
     3541 tctcccaccc catgagctgt cacggtttta tttacatggg gtcaggattc cacgagggta
     3601 gtgaaccatt ttagtcacaa gggcagtggc tgaagatcaa ggagcgggca gtgaactctc
     3661 ctgaatcttc gcctgcttct tcattctcct tcgtttagct aatagaataa ctgctgagtt
     3721 gtgaacagta aggtgtatgt gaggtgctcg aaaacaaggt ttcaggtgac gcccccagaa
     3781 taaaatttgg acggggggtt cagtggtggc attgtgctat gacaccaata taaccctcac
     3841 aaaccccttg ggcaataaat actagtgtag gaatgaaaca ttctgaatat ctttaacaat
     3901 agaaatccat ggggtgggga caagccgtaa agactggatg tccatctcac acgaatttat
     3961 ggctatgggc aacacataat cctagtgcaa tatgatactg gggttattaa gatgtgtccc
     4021 aggcagggac caagacaggt gaaccatgtt gttacactct atttgtaaca aggggaaaga
     4081 gagtggacgc cgacagcagc ggactccact ggttgtctct aacacccccg aaaattaaac
     4141 ggggctccac gccaatgggg cccataaaca aagacaagtg gccactcttt tttttgaaat
     4201 tgtggagtgg gggcacgcgt cagcccccac acgccgccct gcggttttgg actgtaaaat
     4261 aagggtgtaa taacttggct gattgtaacc ccgctaacca ctgcggtcaa accacttgcc
     4321 cacaaaacca ctaatggcac cccggggaat acctgcataa gtaggtgggc gggccaagat
     4381 aggggcgcga ttgctgcgat ctggaggaca aattacacac acttgcgcct gagcgccaag
     4441 cacagggttg ttggtcctca tattcacgag gtcgctgaga gcacggtggg ctaatgttgc
     4501 catgggtagc atatactacc caaatatctg gatagcatat gctatcctaa tctatatctg
     4561 ggtagcatag gctatcctaa tctatatctg ggtagcatat gctatcctaa tctatatctg
     4621 ggtagtatat gctatcctaa tttatatctg ggtagcatag gctatcctaa tctatatctg
     4681 ggtagcatat gctatcctaa tctatatctg ggtagtatat gctatcctaa tctgtatccg
     4741 ggtagcatat gctatcctaa tagagattag ggtagtatat gctatcctaa tttatatctg
     4801 ggtagcatat actacccaaa tatctggata gcatatgcta tcctaatcta tatctgggta
     4861 gcatatgcta tcctaatcta tatctgggta gcataggcta tcctaatcta tatctgggta
     4921 gcatatgcta tcctaatcta tatctgggta gtatatgcta tcctaattta tatctgggta
     4981 gcataggcta tcctaatcta tatctgggta gcatatgcta tcctaatcta tatctgggta
     5041 gtatatgcta tcctaatctg tatccgggta gcatatgcta tcctcatgat aagctgtcaa
     5101 acatgagaat tttcttgaag acgaaagggc ctcgtgatac gcctattttt ataggttaat
     5161 gtcatgataa taatggtttc ttagacgtca ggtggcactt ttcggggaaa tgtgcgcgga
     5221 acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa
     5281 ccctgataaa tgcttcaata atattgaaaa aggaagagta tgagtattca acatttccgt
     5341 gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg
     5401 ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac gagtgggtta catcgaactg
     5461 gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgttt tccaatgatg
     5521 agcactttta aagttctgct atgtggcgcg gtattatccc gtgttgacgc cgggcaagag
     5581 caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtactc accagtcaca
     5641 gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc cataaccatg
     5701 agtgataaca ctgcggccaa cttacttctg acaacgatcg gaggaccgaa ggagctaacc
     5761 gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga accggagctg
     5821 aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgcagcaat ggcaacaacg
     5881 ttgcgcaaac tattaactgg cgaactactt actctagctt cccggcaaca attaatagac
     5941 tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc ggctggctgg
     6001 tttattgctg ataaatctgg agccggtgag cgtgggtctc gcggtatcat tgcagcactg
     6061 gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag tcaggcaact
     6121 atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggtaa
     6181 ctgtcagacc aagtttactc atatatactt tagattgatt taaaacttca tttttaattt
     6241 aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag
     6301 ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct
     6361 ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt
     6421 tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg
     6481 cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt caagaactct
     6541 gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc
     6601 gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg
     6661 tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa
     6721 ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg
     6781 gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg
     6841 ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga
     6901 tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt
     6961 ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct
     7021 gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg ccgcagccga
     7081 acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcccaat acgcaaaccg
     7141 cctctccccg cgcgttggcc gattcattaa tgcagctggc acgacaggtt tcccgactgg
     7201 aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc tcactcatta ggcaccccag
     7261 gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa ttgtgagcgg ataacaattt
     7321 cacacaggaa acagctatga ccatgattac gccaagctct agctagaggt cgaccaattc
     7381 tcatgtttga cagcttatca tcgcagatcc gggcaacgtt gttgccattg ctgcaggcgc
     7441 agaactggta ggtatggaag atctatacat tgaatcaata ttggcaatta gccatattag
     7501 tcattggtta tatagcataa atcaatattg gctattggcc attgcatacg ttgtatctat
     7561 atcataatat gtacatttat attggctcat gtccaatatg accgccatgt tgacattgat
     7621 tattgactag ttattaatag taatcaatta cggggtcatt agttcatagc ccatatatgg
     7681 agttccgcgt tacataactt acggtaaatg gcccgcctgg ctgaccgccc aacgaccccc
     7741 gcccattgac gtcaataatg acgtatgttc ccatagtaac gccaataggg actttccatt
     7801 gacgtcaatg ggtggagtat ttacggtaaa ctgcccactt ggcagtacat caagtgtatc
     7861 atatgccaag tccgccccct attgacgtca atgacggtaa atggcccgcc tggcattatg
     7921 cccagtacat gaccttacgg gactttccta cttggcagta catctacgta ttagtcatcg
     7981 ctattaccat ggtgatgcgg ttttggcagt acaccaatgg gcgtggatag cggtttgact
     8041 cacggggatt tccaagtctc caccccattg acgtcaatgg gagtttgttt tggcaccaaa
     8101 atcaacggga ctttccaaaa tgtcgtaata accccgcccc gttgacgcaa atgggcggta
     8161 ggcgtgtacg gtgggaggtc tatataagca gagctcgttt agtgaaccgt cagatcctca
     8221 ctctcttccg catcgctgtc tgcgagggcc agctgttggg ctcgcggttg aggacaaact
     8281 cttcgcggtc tttccagtac tcttggatcg gaaacccgtc ggcctccgaa cggtactccg
     8341 ccaccgaggg acctgagcga gtccgcatcg accggatcgg aaaacct
//
