LOCUS       sequence-208691-        8809 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 8809)
  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 8809)
  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_0912400-COMP-blac-flag-his
FEATURES             Location/Qualifiers
     source          1..8809
                     /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             2341..3126
                     /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             3133..3198
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             3205..3222
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    3260..3315
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(3314..3333)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(5551..5569)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5637..5741
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5742..5810,5811..6602)
                     /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      6773..7361
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7262..7281
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7515..7532
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    7649..7670
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7685..7690,7691..7708,7709..7715)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7723..7739
                     /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     7728..7750
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7747..7763
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7747..7763
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     enhancer        8034..8413
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        8414..8617
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     8568..8588
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     8614..8638
                     /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 ggccgcccag
      421 ggcgaccagg tcatcaacga gaagctgacc aacttcgtgt tcctgagctg caactaccag
      481 aagggcaaag tcaacaacaa cctgctgaac agcatcgaga agcggaagcc ccagctgatg
      541 ctgtggatcg gcgactactt ctacaccgag tgcagcgaga tcaagtgcct ggacgacgcc
      601 tacacctaca tcaagaaaga ccccttctac atgaagctga agaagaagtt caagatcgac
      661 ggcatctacg acgaccacga ctacaacaag aacaacggcg accggctgta caagtacaag
      721 aaagagtcca agaagaaata cctggactac ctgaacgtgg acaagaacga cgtgcggtac
      781 aagcggaacg gcgcctacat cagcaagctg tacatcgacc ccgacaatga gaagaaccag
      841 gtcaaaatca tcatgctgga cacccggtac aacaaggacc cctacccttt ctacgcccct
      901 gacagctacc gggacctgtt cgtgcacatg ttcatcagct ttctgagccg gttccacagc
      961 agcatcttcg gcctgtgctg caacagcaag aacgatatcc tgggcaacga gcagtggaag
     1021 tggctggaaa gagagctgac aaacagcaac gccagagccc acatcatcat cagctccacc
     1081 cagatcttca gcaaccacat cattaacgag aactggggcc tgatgcccta cagcctgcgg
     1141 cggctgcgcg agctgatcaa aaagaccaag cccaagggcc tgctgttcct gtccggcgac
     1201 gtgcacttcg gctccatcat cggcaaagag gaatccgtca tcgaagtgac cagcagcagc
     1261 gtgaaccagg aaaacatctt cagctacatc aacaaatacg tgatcttctt cctgaccaac
     1321 atcctgagca aggtgtcccc attcgagctg aacaagatct acagcttcaa caatttcggc
     1381 tccgtgaata tcgcctacgt gaacgacaac gagattaaga tcaagacctc cgtgaacgat
     1441 gccgacggcg tggaaatcct ggtggccaat caggtgttca acaataagaa caatatctat
     1501 accaagacca aggatctgca catcatcctg gacgagttcg ccaccctgga atgcaagagc
     1561 aagaccggcg cgccgtcgac ctccatcacg gcctataaga gtgaggggga gtcagcggag
     1621 ttctccttcc cactcaacct tggagaggaa agcctgcagg gagagttgag atggaaggca
     1681 gagaaggctc cttcttccca gtcctggatc accttctccc taaagaacca aaaggtgtct
     1741 gtgcagaagt ctactagcaa ccccaagttc cagctgtccg aaacgctccc actcaccctt
     1801 cagatacccc aggtctccct tcagtttgct ggttctggca acctgaccct gactctggac
     1861 agagggatac tgtatcagga agtgaacctg gtggtgatga aagtgactca gcccgacagc
     1921 aacactttga cctgtgaggt gatgggaccc acctcaccca agatgagact gatcttgaag
     1981 caggagaatc aggaggccag ggtctccagg caggagaaag tgattcaagt gcaggcccct
     2041 gaagcagggg tgtggcaatg tctactgagt gaaggtgaag aggtcaagat ggactccaag
     2101 atccaggttt tatccaaagg gttgaattcc ggcggcggct ccggaggcgg taccggtggc
     2161 ggcgacctag ccccacagat gcttcgagaa ctccaggaga ctaatgcggc gctgcaagac
     2221 gtgagagagc tcttgcgaca gcaggtcaag gagatcacct tcctgaagaa tacggtgatg
     2281 gaatgtgacg cctgcggtct agacagaaac ctgcctccac tagcacctct gggcccacac
     2341 ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg agtgggttac
     2401 atcgaactgg atctcaacag cggtaagatc cttgagagtt ttcgccccga agaacgtttt
     2461 ccaatgatga gcacttttaa agttctgcta tgtggcgcgg tattatcccg tattgacgcc
     2521 gggcaagagc aactcggtcg ccgcatacac tattctcaga atgacttggt tgagtactca
     2581 ccagtcacag aaaagcatct tacggatggc atgacagtaa gagaattatg cagtgctgcc
     2641 ataaccatga gtgataacac tgcggccaac ttacttctga caacgatcgg aggaccgaag
     2701 gagctaaccg cttttttgca caacatgggg gatcatgtaa ctcgccttga tcgttgggaa
     2761 ccggagctga atgaagccat accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg
     2821 gcaacaacgt tgcgcaaact attaactggc gaactactta ctctagcttc ccggcaacaa
     2881 ttaatagact ggatggaggc ggataaagtt gcaggaccac ttctgcgctc ggcccttccg
     2941 gctggctggt ttattgctga taaatctgga gccggtgagc gtgggtctcg cggtatcatt
     3001 gcagcactgg ggccagatgg taagccctcc cgtatcgtag ttatctacac gacggggagt
     3061 caggcaacta tggatgaacg aaatagacag atcgctgaga taggtgcctc actgattaag
     3121 cattggggcc ctgactacaa agaccatgac ggtgattata aagatcatga catcgattac
     3181 aaggatgacg atgacaaggg ccctcaccac caccatcacc actaattaat taaggatccc
     3241 ccgacctcga cctctggcta ataaaggaaa tttattttca ttgcaatagt gtgttggaat
     3301 tttttgtgtc tctcactcgg aaggacatat gggagggcaa atcatttggt cgagatccct
     3361 cggagatctc tagctagagg atcgatcccc gccccggacg aactaaacct gactacgaca
     3421 tctctgcccc ttcttcgcgg ggcagtgcat gtaatccctt cagttggttg gtacaacttg
     3481 ccaactgggc cctgttccac atgtgacacg gggggggacc aaacacaaag gggttctctg
     3541 actgtagttg acatccttat aaatggatgt gcacatttgc caacactgag tggctttcat
     3601 cctggagcag actttgcagt ctgtggactg caacacaaca ttgcctttat gtgtaactct
     3661 tggctgaagc tcttacacca atgctggggg acatgtacct cccaggggcc caggaagact
     3721 acgggaggct acaccaacgt caatcagagg ggcctgtgta gctaccgata agcggaccct
     3781 caagagggca ttagcaatag tgtttataag gcccccttgt taaccctaaa cgggtagcat
     3841 atgcttcccg ggtagtagta tatactatcc agactaaccc taattcaata gcatatgtta
     3901 cccaacggga agcatatgct atcgaattag ggttagtaaa agggtcctaa ggaacagcga
     3961 tatctcccac cccatgagct gtcacggttt tatttacatg gggtcaggat tccacgaggg
     4021 tagtgaacca ttttagtcac aagggcagtg gctgaagatc aaggagcggg cagtgaactc
     4081 tcctgaatct tcgcctgctt cttcattctc cttcgtttag ctaatagaat aactgctgag
     4141 ttgtgaacag taaggtgtat gtgaggtgct cgaaaacaag gtttcaggtg acgcccccag
     4201 aataaaattt ggacgggggg ttcagtggtg gcattgtgct atgacaccaa tataaccctc
     4261 acaaacccct tgggcaataa atactagtgt aggaatgaaa cattctgaat atctttaaca
     4321 atagaaatcc atggggtggg gacaagccgt aaagactgga tgtccatctc acacgaattt
     4381 atggctatgg gcaacacata atcctagtgc aatatgatac tggggttatt aagatgtgtc
     4441 ccaggcaggg accaagacag gtgaaccatg ttgttacact ctatttgtaa caaggggaaa
     4501 gagagtggac gccgacagca gcggactcca ctggttgtct ctaacacccc cgaaaattaa
     4561 acggggctcc acgccaatgg ggcccataaa caaagacaag tggccactct tttttttgaa
     4621 attgtggagt gggggcacgc gtcagccccc acacgccgcc ctgcggtttt ggactgtaaa
     4681 ataagggtgt aataacttgg ctgattgtaa ccccgctaac cactgcggtc aaaccacttg
     4741 cccacaaaac cactaatggc accccgggga atacctgcat aagtaggtgg gcgggccaag
     4801 ataggggcgc gattgctgcg atctggagga caaattacac acacttgcgc ctgagcgcca
     4861 agcacagggt tgttggtcct catattcacg aggtcgctga gagcacggtg ggctaatgtt
     4921 gccatgggta gcatatacta cccaaatatc tggatagcat atgctatcct aatctatatc
     4981 tgggtagcat aggctatcct aatctatatc tgggtagcat atgctatcct aatctatatc
     5041 tgggtagtat atgctatcct aatttatatc tgggtagcat aggctatcct aatctatatc
     5101 tgggtagcat atgctatcct aatctatatc tgggtagtat atgctatcct aatctgtatc
     5161 cgggtagcat atgctatcct aatagagatt agggtagtat atgctatcct aatttatatc
     5221 tgggtagcat atactaccca aatatctgga tagcatatgc tatcctaatc tatatctggg
     5281 tagcatatgc tatcctaatc tatatctggg tagcataggc tatcctaatc tatatctggg
     5341 tagcatatgc tatcctaatc tatatctggg tagtatatgc tatcctaatt tatatctggg
     5401 tagcataggc tatcctaatc tatatctggg tagcatatgc tatcctaatc tatatctggg
     5461 tagtatatgc tatcctaatc tgtatccggg tagcatatgc tatcctcatg ataagctgtc
     5521 aaacatgaga attttcttga agacgaaagg gcctcgtgat acgcctattt ttataggtta
     5581 atgtcatgat aataatggtt tcttagacgt caggtggcac ttttcgggga aatgtgcgcg
     5641 gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat
     5701 aaccctgata aatgcttcaa taatattgaa aaaggaagag tatgagtatt caacatttcc
     5761 gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct cacccagaaa
     5821 cgctggtgaa agtaaaagat gctgaagatc agttgggtgc acgagtgggt tacatcgaac
     5881 tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgt tttccaatga
     5941 tgagcacttt taaagttctg ctatgtggcg cggtattatc ccgtgttgac gccgggcaag
     6001 agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtac tcaccagtca
     6061 cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct gccataacca
     6121 tgagtgataa cactgcggcc aacttacttc tgacaacgat cggaggaccg aaggagctaa
     6181 ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg gaaccggagc
     6241 tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgcagca atggcaacaa
     6301 cgttgcgcaa actattaact ggcgaactac ttactctagc ttcccggcaa caattaatag
     6361 actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt ccggctggct
     6421 ggtttattgc tgataaatct ggagccggtg agcgtgggtc tcgcggtatc attgcagcac
     6481 tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg agtcaggcaa
     6541 ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt aagcattggt
     6601 aactgtcaga ccaagtttac tcatatatac tttagattga tttaaaactt catttttaat
     6661 ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg
     6721 agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc
     6781 ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg
     6841 tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag
     6901 cgcagatacc aaatactgtt cttctagtgt agccgtagtt aggccaccac ttcaagaact
     6961 ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg
     7021 gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc
     7081 ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg
     7141 aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg
     7201 cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag
     7261 ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc
     7321 gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct
     7381 ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct gcgttatccc
     7441 ctgattctgt ggataaccgt attaccgcct ttgagtgagc tgataccgct cgccgcagcc
     7501 gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga agagcgccca atacgcaaac
     7561 cgcctctccc cgcgcgttgg ccgattcatt aatgcagctg gcacgacagg tttcccgact
     7621 ggaaagcggg cagtgagcgc aacgcaatta atgtgagtta gctcactcat taggcacccc
     7681 aggctttaca ctttatgctt ccggctcgta tgttgtgtgg aattgtgagc ggataacaat
     7741 ttcacacagg aaacagctat gaccatgatt acgccaagct ctagctagag gtcgaccaat
     7801 tctcatgttt gacagcttat catcgcagat ccgggcaacg ttgttgccat tgctgcaggc
     7861 gcagaactgg taggtatgga agatctatac attgaatcaa tattggcaat tagccatatt
     7921 agtcattggt tatatagcat aaatcaatat tggctattgg ccattgcata cgttgtatct
     7981 atatcataat atgtacattt atattggctc atgtccaata tgaccgccat gttgacattg
     8041 attattgact agttattaat agtaatcaat tacggggtca ttagttcata gcccatatat
     8101 ggagttccgc gttacataac ttacggtaaa tggcccgcct ggctgaccgc ccaacgaccc
     8161 ccgcccattg acgtcaataa tgacgtatgt tcccatagta acgccaatag ggactttcca
     8221 ttgacgtcaa tgggtggagt atttacggta aactgcccac ttggcagtac atcaagtgta
     8281 tcatatgcca agtccgcccc ctattgacgt caatgacggt aaatggcccg cctggcatta
     8341 tgcccagtac atgaccttac gggactttcc tacttggcag tacatctacg tattagtcat
     8401 cgctattacc atggtgatgc ggttttggca gtacaccaat gggcgtggat agcggtttga
     8461 ctcacgggga tttccaagtc tccaccccat tgacgtcaat gggagtttgt tttggcacca
     8521 aaatcaacgg gactttccaa aatgtcgtaa taaccccgcc ccgttgacgc aaatgggcgg
     8581 taggcgtgta cggtgggagg tctatataag cagagctcgt ttagtgaacc gtcagatcct
     8641 cactctcttc cgcatcgctg tctgcgaggg ccagctgttg ggctcgcggt tgaggacaaa
     8701 ctcttcgcgg tctttccagt actcttggat cggaaacccg tcggcctccg aacggtactc
     8761 cgccaccgag ggacctgagc gagtccgcat cgaccggatc ggaaaacct
//
