LOCUS       sequence-208710-        8389 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 8389)
  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 8389)
  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..8389
                     /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    2840..2895
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(2894..2913)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(5131..5149)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5217..5321
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5322..5390,5391..6182)
                     /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      6353..6941
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6842..6861
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7095..7112
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    7229..7250
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7265..7270,7271..7288,7289..7295)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7303..7319
                     /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     7308..7330
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7327..7343
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7327..7343
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     enhancer        7614..7993
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        7994..8197
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     8148..8168
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     8194..8218
                     /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 actaattaat taaggatccc
     2821 ccgacctcga cctctggcta ataaaggaaa tttattttca ttgcaatagt gtgttggaat
     2881 tttttgtgtc tctcactcgg aaggacatat gggagggcaa atcatttggt cgagatccct
     2941 cggagatctc tagctagagg atcgatcccc gccccggacg aactaaacct gactacgaca
     3001 tctctgcccc ttcttcgcgg ggcagtgcat gtaatccctt cagttggttg gtacaacttg
     3061 ccaactgggc cctgttccac atgtgacacg gggggggacc aaacacaaag gggttctctg
     3121 actgtagttg acatccttat aaatggatgt gcacatttgc caacactgag tggctttcat
     3181 cctggagcag actttgcagt ctgtggactg caacacaaca ttgcctttat gtgtaactct
     3241 tggctgaagc tcttacacca atgctggggg acatgtacct cccaggggcc caggaagact
     3301 acgggaggct acaccaacgt caatcagagg ggcctgtgta gctaccgata agcggaccct
     3361 caagagggca ttagcaatag tgtttataag gcccccttgt taaccctaaa cgggtagcat
     3421 atgcttcccg ggtagtagta tatactatcc agactaaccc taattcaata gcatatgtta
     3481 cccaacggga agcatatgct atcgaattag ggttagtaaa agggtcctaa ggaacagcga
     3541 tatctcccac cccatgagct gtcacggttt tatttacatg gggtcaggat tccacgaggg
     3601 tagtgaacca ttttagtcac aagggcagtg gctgaagatc aaggagcggg cagtgaactc
     3661 tcctgaatct tcgcctgctt cttcattctc cttcgtttag ctaatagaat aactgctgag
     3721 ttgtgaacag taaggtgtat gtgaggtgct cgaaaacaag gtttcaggtg acgcccccag
     3781 aataaaattt ggacgggggg ttcagtggtg gcattgtgct atgacaccaa tataaccctc
     3841 acaaacccct tgggcaataa atactagtgt aggaatgaaa cattctgaat atctttaaca
     3901 atagaaatcc atggggtggg gacaagccgt aaagactgga tgtccatctc acacgaattt
     3961 atggctatgg gcaacacata atcctagtgc aatatgatac tggggttatt aagatgtgtc
     4021 ccaggcaggg accaagacag gtgaaccatg ttgttacact ctatttgtaa caaggggaaa
     4081 gagagtggac gccgacagca gcggactcca ctggttgtct ctaacacccc cgaaaattaa
     4141 acggggctcc acgccaatgg ggcccataaa caaagacaag tggccactct tttttttgaa
     4201 attgtggagt gggggcacgc gtcagccccc acacgccgcc ctgcggtttt ggactgtaaa
     4261 ataagggtgt aataacttgg ctgattgtaa ccccgctaac cactgcggtc aaaccacttg
     4321 cccacaaaac cactaatggc accccgggga atacctgcat aagtaggtgg gcgggccaag
     4381 ataggggcgc gattgctgcg atctggagga caaattacac acacttgcgc ctgagcgcca
     4441 agcacagggt tgttggtcct catattcacg aggtcgctga gagcacggtg ggctaatgtt
     4501 gccatgggta gcatatacta cccaaatatc tggatagcat atgctatcct aatctatatc
     4561 tgggtagcat aggctatcct aatctatatc tgggtagcat atgctatcct aatctatatc
     4621 tgggtagtat atgctatcct aatttatatc tgggtagcat aggctatcct aatctatatc
     4681 tgggtagcat atgctatcct aatctatatc tgggtagtat atgctatcct aatctgtatc
     4741 cgggtagcat atgctatcct aatagagatt agggtagtat atgctatcct aatttatatc
     4801 tgggtagcat atactaccca aatatctgga tagcatatgc tatcctaatc tatatctggg
     4861 tagcatatgc tatcctaatc tatatctggg tagcataggc tatcctaatc tatatctggg
     4921 tagcatatgc tatcctaatc tatatctggg tagtatatgc tatcctaatt tatatctggg
     4981 tagcataggc tatcctaatc tatatctggg tagcatatgc tatcctaatc tatatctggg
     5041 tagtatatgc tatcctaatc tgtatccggg tagcatatgc tatcctcatg ataagctgtc
     5101 aaacatgaga attttcttga agacgaaagg gcctcgtgat acgcctattt ttataggtta
     5161 atgtcatgat aataatggtt tcttagacgt caggtggcac ttttcgggga aatgtgcgcg
     5221 gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat
     5281 aaccctgata aatgcttcaa taatattgaa aaaggaagag tatgagtatt caacatttcc
     5341 gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct cacccagaaa
     5401 cgctggtgaa agtaaaagat gctgaagatc agttgggtgc acgagtgggt tacatcgaac
     5461 tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgt tttccaatga
     5521 tgagcacttt taaagttctg ctatgtggcg cggtattatc ccgtgttgac gccgggcaag
     5581 agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtac tcaccagtca
     5641 cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct gccataacca
     5701 tgagtgataa cactgcggcc aacttacttc tgacaacgat cggaggaccg aaggagctaa
     5761 ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg gaaccggagc
     5821 tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgcagca atggcaacaa
     5881 cgttgcgcaa actattaact ggcgaactac ttactctagc ttcccggcaa caattaatag
     5941 actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt ccggctggct
     6001 ggtttattgc tgataaatct ggagccggtg agcgtgggtc tcgcggtatc attgcagcac
     6061 tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg agtcaggcaa
     6121 ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt aagcattggt
     6181 aactgtcaga ccaagtttac tcatatatac tttagattga tttaaaactt catttttaat
     6241 ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg
     6301 agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc
     6361 ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg
     6421 tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag
     6481 cgcagatacc aaatactgtt cttctagtgt agccgtagtt aggccaccac ttcaagaact
     6541 ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg
     6601 gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc
     6661 ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg
     6721 aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg
     6781 cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag
     6841 ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc
     6901 gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct
     6961 ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct gcgttatccc
     7021 ctgattctgt ggataaccgt attaccgcct ttgagtgagc tgataccgct cgccgcagcc
     7081 gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga agagcgccca atacgcaaac
     7141 cgcctctccc cgcgcgttgg ccgattcatt aatgcagctg gcacgacagg tttcccgact
     7201 ggaaagcggg cagtgagcgc aacgcaatta atgtgagtta gctcactcat taggcacccc
     7261 aggctttaca ctttatgctt ccggctcgta tgttgtgtgg aattgtgagc ggataacaat
     7321 ttcacacagg aaacagctat gaccatgatt acgccaagct ctagctagag gtcgaccaat
     7381 tctcatgttt gacagcttat catcgcagat ccgggcaacg ttgttgccat tgctgcaggc
     7441 gcagaactgg taggtatgga agatctatac attgaatcaa tattggcaat tagccatatt
     7501 agtcattggt tatatagcat aaatcaatat tggctattgg ccattgcata cgttgtatct
     7561 atatcataat atgtacattt atattggctc atgtccaata tgaccgccat gttgacattg
     7621 attattgact agttattaat agtaatcaat tacggggtca ttagttcata gcccatatat
     7681 ggagttccgc gttacataac ttacggtaaa tggcccgcct ggctgaccgc ccaacgaccc
     7741 ccgcccattg acgtcaataa tgacgtatgt tcccatagta acgccaatag ggactttcca
     7801 ttgacgtcaa tgggtggagt atttacggta aactgcccac ttggcagtac atcaagtgta
     7861 tcatatgcca agtccgcccc ctattgacgt caatgacggt aaatggcccg cctggcatta
     7921 tgcccagtac atgaccttac gggactttcc tacttggcag tacatctacg tattagtcat
     7981 cgctattacc atggtgatgc ggttttggca gtacaccaat gggcgtggat agcggtttga
     8041 ctcacgggga tttccaagtc tccaccccat tgacgtcaat gggagtttgt tttggcacca
     8101 aaatcaacgg gactttccaa aatgtcgtaa taaccccgcc ccgttgacgc aaatgggcgg
     8161 taggcgtgta cggtgggagg tctatataag cagagctcgt ttagtgaacc gtcagatcct
     8221 cactctcttc cgcatcgctg tctgcgaggg ccagctgttg ggctcgcggt tgaggacaaa
     8281 ctcttcgcgg tctttccagt actcttggat cggaaacccg tcggcctccg aacggtactc
     8341 cgccaccgag ggacctgagc gagtccgcat cgaccggatc ggaaaacct
//
