LOCUS       sequence-224704-        8789 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 8789)
  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 8789)
  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_0107300-COMP-blac-flag-his
FEATURES             Location/Qualifiers
     source          1..8789
                     /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             2323..3108
                     /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             3115..3180
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             3187..3204
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    3240..3295
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(3294..3313)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(5531..5549)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5617..5721
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5722..5790,5791..6582)
                     /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      6753..7341
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7242..7261
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7495..7512
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    7629..7650
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7665..7670,7671..7688,7689..7695)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7703..7719
                     /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     7708..7730
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7727..7743
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7727..7743
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     enhancer        8014..8393
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        8394..8597
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     8548..8568
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     8594..8618
                     /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 ggccgccatc
      421 aaccctgcca ccctgatcca gaaccacctg tacatgtatc agaccaacaa cgccatcaat
      481 gaagagaaca tctacaacat gaagaagttc agcaagctgc ctatcgccga gaacacccgg
      541 aacatccggc acagcaccaa cgacctgtac tacgagcaca gcctgaacct ggaaaagccc
      601 aacgagcact tcaccctgca caacatcaag ctgatctgcc agcagatcta ccacaaccag
      661 cccaaccagc tgaacctgcg gatcctggcc gtgctgctga gcaccagcta caagcaggaa
      721 aacattgcca acatcttcca gaacctgtgc aacaacggcc tgagctgcct gttcaccatc
      781 cagctgagca agaacaaggc cttcgtggac cagaacaaga tcgtgcacct gaagatcatc
      841 gacaccgacg acctgaagcg gaacaagaac aacaacatca ggcacaagat caccgtgtac
      901 tacatctgcg aggacctgaa gtacaagaac cagaagcagc aggaaagcaa cgagttcaaa
      961 tacgtgttca attacgagca gctggccttc ctgaagtgcg agaactacct gaccgtgcag
     1021 gtcgagaagc tcgtgggcat caactacaac aaagagttcc acaacctgaa ccccgagtac
     1081 atcagcttct actgcagcca gagagtgatc tgcaacaccc acatcgagga tctgtacaaa
     1141 gaggacgaca agaagcggaa gaactacatc ttctacgagc tgacctaccg ctgtgtgcct
     1201 cccaaagtgt gcagcttctg cggcaacaat gccgcctgca agctggaaaa actgtataac
     1261 aaagagaaag agatcaccca gttctacaac aacgtcgagt acaacgatgc cggcgtgttc
     1321 gaccggctgc tgctgaacaa agaagaggaa gaggaacgca acaaagccag cttcgtggcc
     1381 aacctcgagt ataacgccat taactacgac atcgacgaca tgatcacagt gggcagcaac
     1441 acctaccaca tcctggaaat ctgcaagtgc agcaaggact acgtgcagga aggctcctcc
     1501 tgcgtcaaga aagagaagga caagatctac atccgctacg ccaagaaagg cgcgccgtcg
     1561 acctccatca cggcctataa gagtgagggg gagtcagcgg agttctcctt cccactcaac
     1621 cttggagagg aaagcctgca gggagagttg agatggaagg cagagaaggc tccttcttcc
     1681 cagtcctgga tcaccttctc cctaaagaac caaaaggtgt ctgtgcagaa gtctactagc
     1741 aaccccaagt tccagctgtc cgaaacgctc ccactcaccc ttcagatacc ccaggtctcc
     1801 cttcagtttg ctggttctgg caacctgacc ctgactctgg acagagggat actgtatcag
     1861 gaagtgaacc tggtggtgat gaaagtgact cagcccgaca gcaacacttt gacctgtgag
     1921 gtgatgggac ccacctcacc caagatgaga ctgatcttga agcaggagaa tcaggaggcc
     1981 agggtctcca ggcaggagaa agtgattcaa gtgcaggccc ctgaagcagg ggtgtggcaa
     2041 tgtctactga gtgaaggtga agaggtcaag atggactcca agatccaggt tttatccaaa
     2101 gggttgaatt ccggcggcgg ctccggaggc ggtaccggtg gcggcgacct agccccacag
     2161 atgcttcgag aactccagga gactaatgcg gcgctgcaag acgtgagaga gctcttgcga
     2221 cagcaggtca aggagatcac cttcctgaag aatacggtga tggaatgtga cgcctgcggt
     2281 ctagacagaa acctgcctcc actagcacct ctgggcccac acccagaaac gctggtgaaa
     2341 gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac
     2401 agcggtaaga tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt
     2461 aaagttctgc tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga gcaactcggt
     2521 cgccgcatac actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat
     2581 cttacggatg gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac
     2641 actgcggcca acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg
     2701 cacaacatgg gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc
     2761 ataccaaacg acgagcgtga caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa
     2821 ctattaactg gcgaactact tactctagct tcccggcaac aattaataga ctggatggag
     2881 gcggataaag ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct
     2941 gataaatctg gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat
     3001 ggtaagccct cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa
     3061 cgaaatagac agatcgctga gataggtgcc tcactgatta agcattgggg ccctgactac
     3121 aaagaccatg acggtgatta taaagatcat gacatcgatt acaaggatga cgatgacaag
     3181 ggccctcacc accaccatca ccactaataa taaggatccc ccgacctcga cctctggcta
     3241 ataaaggaaa tttattttca ttgcaatagt gtgttggaat tttttgtgtc tctcactcgg
     3301 aaggacatat gggagggcaa atcatttggt cgagatccct cggagatctc tagctagagg
     3361 atcgatcccc gccccggacg aactaaacct gactacgaca tctctgcccc ttcttcgcgg
     3421 ggcagtgcat gtaatccctt cagttggttg gtacaacttg ccaactgggc cctgttccac
     3481 atgtgacacg gggggggacc aaacacaaag gggttctctg actgtagttg acatccttat
     3541 aaatggatgt gcacatttgc caacactgag tggctttcat cctggagcag actttgcagt
     3601 ctgtggactg caacacaaca ttgcctttat gtgtaactct tggctgaagc tcttacacca
     3661 atgctggggg acatgtacct cccaggggcc caggaagact acgggaggct acaccaacgt
     3721 caatcagagg ggcctgtgta gctaccgata agcggaccct caagagggca ttagcaatag
     3781 tgtttataag gcccccttgt taaccctaaa cgggtagcat atgcttcccg ggtagtagta
     3841 tatactatcc agactaaccc taattcaata gcatatgtta cccaacggga agcatatgct
     3901 atcgaattag ggttagtaaa agggtcctaa ggaacagcga tatctcccac cccatgagct
     3961 gtcacggttt tatttacatg gggtcaggat tccacgaggg tagtgaacca ttttagtcac
     4021 aagggcagtg gctgaagatc aaggagcggg cagtgaactc tcctgaatct tcgcctgctt
     4081 cttcattctc cttcgtttag ctaatagaat aactgctgag ttgtgaacag taaggtgtat
     4141 gtgaggtgct cgaaaacaag gtttcaggtg acgcccccag aataaaattt ggacgggggg
     4201 ttcagtggtg gcattgtgct atgacaccaa tataaccctc acaaacccct tgggcaataa
     4261 atactagtgt aggaatgaaa cattctgaat atctttaaca atagaaatcc atggggtggg
     4321 gacaagccgt aaagactgga tgtccatctc acacgaattt atggctatgg gcaacacata
     4381 atcctagtgc aatatgatac tggggttatt aagatgtgtc ccaggcaggg accaagacag
     4441 gtgaaccatg ttgttacact ctatttgtaa caaggggaaa gagagtggac gccgacagca
     4501 gcggactcca ctggttgtct ctaacacccc cgaaaattaa acggggctcc acgccaatgg
     4561 ggcccataaa caaagacaag tggccactct tttttttgaa attgtggagt gggggcacgc
     4621 gtcagccccc acacgccgcc ctgcggtttt ggactgtaaa ataagggtgt aataacttgg
     4681 ctgattgtaa ccccgctaac cactgcggtc aaaccacttg cccacaaaac cactaatggc
     4741 accccgggga atacctgcat aagtaggtgg gcgggccaag ataggggcgc gattgctgcg
     4801 atctggagga caaattacac acacttgcgc ctgagcgcca agcacagggt tgttggtcct
     4861 catattcacg aggtcgctga gagcacggtg ggctaatgtt gccatgggta gcatatacta
     4921 cccaaatatc tggatagcat atgctatcct aatctatatc tgggtagcat aggctatcct
     4981 aatctatatc tgggtagcat atgctatcct aatctatatc tgggtagtat atgctatcct
     5041 aatttatatc tgggtagcat aggctatcct aatctatatc tgggtagcat atgctatcct
     5101 aatctatatc tgggtagtat atgctatcct aatctgtatc cgggtagcat atgctatcct
     5161 aatagagatt agggtagtat atgctatcct aatttatatc tgggtagcat atactaccca
     5221 aatatctgga tagcatatgc tatcctaatc tatatctggg tagcatatgc tatcctaatc
     5281 tatatctggg tagcataggc tatcctaatc tatatctggg tagcatatgc tatcctaatc
     5341 tatatctggg tagtatatgc tatcctaatt tatatctggg tagcataggc tatcctaatc
     5401 tatatctggg tagcatatgc tatcctaatc tatatctggg tagtatatgc tatcctaatc
     5461 tgtatccggg tagcatatgc tatcctcatg ataagctgtc aaacatgaga attttcttga
     5521 agacgaaagg gcctcgtgat acgcctattt ttataggtta atgtcatgat aataatggtt
     5581 tcttagacgt caggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt
     5641 ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa
     5701 taatattgaa aaaggaagag tatgagtatt caacatttcc gtgtcgccct tattcccttt
     5761 tttgcggcat tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat
     5821 gctgaagatc agttgggtgc acgagtgggt tacatcgaac tggatctcaa cagcggtaag
     5881 atccttgaga gttttcgccc cgaagaacgt tttccaatga tgagcacttt taaagttctg
     5941 ctatgtggcg cggtattatc ccgtgttgac gccgggcaag agcaactcgg tcgccgcata
     6001 cactattctc agaatgactt ggttgagtac tcaccagtca cagaaaagca tcttacggat
     6061 ggcatgacag taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc
     6121 aacttacttc tgacaacgat cggaggaccg aaggagctaa ccgctttttt gcacaacatg
     6181 ggggatcatg taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac
     6241 gacgagcgtg acaccacgat gcctgcagca atggcaacaa cgttgcgcaa actattaact
     6301 ggcgaactac ttactctagc ttcccggcaa caattaatag actggatgga ggcggataaa
     6361 gttgcaggac cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct
     6421 ggagccggtg agcgtgggtc tcgcggtatc attgcagcac tggggccaga tggtaagccc
     6481 tcccgtatcg tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga
     6541 cagatcgctg agataggtgc ctcactgatt aagcattggt aactgtcaga ccaagtttac
     6601 tcatatatac tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag
     6661 atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg
     6721 tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc
     6781 tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag
     6841 ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtt
     6901 cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac
     6961 ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc
     7021 gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt
     7081 tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt
     7141 gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc
     7201 ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt
     7261 tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca
     7321 ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt
     7381 tgctggcctt ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt
     7441 attaccgcct ttgagtgagc tgataccgct cgccgcagcc gaacgaccga gcgcagcgag
     7501 tcagtgagcg aggaagcgga agagcgccca atacgcaaac cgcctctccc cgcgcgttgg
     7561 ccgattcatt aatgcagctg gcacgacagg tttcccgact ggaaagcggg cagtgagcgc
     7621 aacgcaatta atgtgagtta gctcactcat taggcacccc aggctttaca ctttatgctt
     7681 ccggctcgta tgttgtgtgg aattgtgagc ggataacaat ttcacacagg aaacagctat
     7741 gaccatgatt acgccaagct ctagctagag gtcgaccaat tctcatgttt gacagcttat
     7801 catcgcagat ccgggcaacg ttgttgccat tgctgcaggc gcagaactgg taggtatgga
     7861 agatctatac attgaatcaa tattggcaat tagccatatt agtcattggt tatatagcat
     7921 aaatcaatat tggctattgg ccattgcata cgttgtatct atatcataat atgtacattt
     7981 atattggctc atgtccaata tgaccgccat gttgacattg attattgact agttattaat
     8041 agtaatcaat tacggggtca ttagttcata gcccatatat ggagttccgc gttacataac
     8101 ttacggtaaa tggcccgcct ggctgaccgc ccaacgaccc ccgcccattg acgtcaataa
     8161 tgacgtatgt tcccatagta acgccaatag ggactttcca ttgacgtcaa tgggtggagt
     8221 atttacggta aactgcccac ttggcagtac atcaagtgta tcatatgcca agtccgcccc
     8281 ctattgacgt caatgacggt aaatggcccg cctggcatta tgcccagtac atgaccttac
     8341 gggactttcc tacttggcag tacatctacg tattagtcat cgctattacc atggtgatgc
     8401 ggttttggca gtacaccaat gggcgtggat agcggtttga ctcacgggga tttccaagtc
     8461 tccaccccat tgacgtcaat gggagtttgt tttggcacca aaatcaacgg gactttccaa
     8521 aatgtcgtaa taaccccgcc ccgttgacgc aaatgggcgg taggcgtgta cggtgggagg
     8581 tctatataag cagagctcgt ttagtgaacc gtcagatcct cactctcttc cgcatcgctg
     8641 tctgcgaggg ccagctgttg ggctcgcggt tgaggacaaa ctcttcgcgg tctttccagt
     8701 actcttggat cggaaacccg tcggcctccg aacggtactc cgccaccgag ggacctgagc
     8761 gagtccgcat cgaccggatc ggaaaacct
//
