LOCUS       sequence-223566-        9629 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 9629)
  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 9629)
  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_1105300-COMP-blac-flag-his
FEATURES             Location/Qualifiers
     source          1..9629
                     /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             3163..3948
                     /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             3955..4020
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             4027..4044
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    4080..4135
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(4134..4153)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(6371..6389)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6457..6561
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6562..6630,6631..7422)
                     /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      7593..8181
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8082..8101
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     8335..8352
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    8469..8490
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(8505..8510,8511..8528,8529..8535)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8543..8559
                     /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     8548..8570
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     8567..8583
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     8567..8583
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     enhancer        8854..9233
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        9234..9437
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     9388..9408
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     9434..9458
                     /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 ggccgcctac
      421 aagtacaccg tggaaggcaa agaagtgaaa gtcatctaca ccaacgacgc caacaagaac
      481 aagaagaaaa acgagctgaa caacttccag tacatcttcc tgaacgagaa cgaggaaaac
      541 tacaatatcg ccagcagcga cctgaacgac atcaacatcg aggaagagga catctacttc
      601 tacaagagct tcaccaagat catcaactac ttcaagtacc ggatccccgc caacatcaac
      661 ttcgccttcg agtacaacga cggcgacatc atcctgctgg acaaaaagaa aaagggcgtg
      721 tcctgcaaga tcaacaacct gttctacagc gagagcggcc tgaagtacct ggaccggaag
      781 aacgagctgt acgtgaagtc cgaggccttc gtgatcaagc acggcaacca ggaaaacaac
      841 gctaacaacg acaagaacga gggcatgaac caccggaaca acaataataa caataacaac
      901 tacaacaaca attataacaa caacaacaat tacaacaact actacaacaa cgccttcaac
      961 agcgccgtgc gcgaggacga gttcatcagc ggcaccgccc tgtgcgagaa cagaaacaag
     1021 aactacctgc ccaagaagat gctctgcttc cccagcagca acaaaaacaa tgagctgaat
     1081 atcgccttca tcaagaacga agagtacaaa gaactgatgg aactgtacat gaccggcatc
     1141 aagaaggaca acacctatct gctgaagaag atcgccaatt acttcaagag catcctgaac
     1201 ctggactacg acgtgaagaa tattgcctac tacatcttct gcaagaccct gcagtgcaac
     1261 ctgcagaacg tgttcgtgct gccctacgag tctgaggaag agtataagat ccagaacatc
     1321 atctggtcct ataacaacga gcagatcgcc ctgaacaatg ccttcatgta caacaatatc
     1381 ctggggaacg agaagatcgg caaccggaat atcatctggc ggcacgagaa taacaacatc
     1441 gtgaagcaca acattgccaa gtgcaacagc atctttttca acgagatgga atattacaac
     1501 tgctacaaga acctcgagaa cgtgtacgag atcctgctct acaaagagac aaacgtcctg
     1561 aacaagtaca tcaccaacaa ggccatcatt aagaacattg cctccgtgaa ccagtacacc
     1621 gccatcttct acgacgacaa cgactatgtg atcctgctga aggattacaa gtactacatc
     1681 agcttccagc tgaagaacat tatcgacctg gtgctgttcc aggacagcat caatgagatc
     1741 agcttctact acattaactt cgagagcaac aacatgctgc tgcggatcta caagtgcgcc
     1801 attaacaaca atgaagtgaa gtgcgccgac atcagctaca tcccttacga gatcaacgcc
     1861 aaagagaagt tcatctacct gagcaccaac cagaaaaagg atgtgtccag tgtgttcgtg
     1921 tccaccaaca ccaaagtgtg gaagatctat cacaccaagc aggaaaacga caacaacaac
     1981 aacaataaca ataacaacaa caacaataat aacaacaata acaataacaa caacaataat
     2041 aacaataaca acaacaataa caatgataac aatctgtaca gccgcaagct gatcgacagc
     2101 atccagaata ccaacgagga atacttcggc cacatcaatt gctacggcat cgaggccacc
     2161 tacgagagaa agctgttcag caagtatctg aacaacatcc ccgacgataa caagaacgtc
     2221 gtgggctgca gctacctgaa gtattacatc aatagcgcct ccaaggccct gctgatctcc
     2281 ttcatcgaga atgcccactt catccagcgg acctaccacg tcgtgaaaaa caagaatatc
     2341 attgtgtcct ccggcaccac aaagctgatc atcagagtga atgagctggg cgcgccgtcg
     2401 acctccatca cggcctataa gagtgagggg gagtcagcgg agttctcctt cccactcaac
     2461 cttggagagg aaagcctgca gggagagttg agatggaagg cagagaaggc tccttcttcc
     2521 cagtcctgga tcaccttctc cctaaagaac caaaaggtgt ctgtgcagaa gtctactagc
     2581 aaccccaagt tccagctgtc cgaaacgctc ccactcaccc ttcagatacc ccaggtctcc
     2641 cttcagtttg ctggttctgg caacctgacc ctgactctgg acagagggat actgtatcag
     2701 gaagtgaacc tggtggtgat gaaagtgact cagcccgaca gcaacacttt gacctgtgag
     2761 gtgatgggac ccacctcacc caagatgaga ctgatcttga agcaggagaa tcaggaggcc
     2821 agggtctcca ggcaggagaa agtgattcaa gtgcaggccc ctgaagcagg ggtgtggcaa
     2881 tgtctactga gtgaaggtga agaggtcaag atggactcca agatccaggt tttatccaaa
     2941 gggttgaatt ccggcggcgg ctccggaggc ggtaccggtg gcggcgacct agccccacag
     3001 atgcttcgag aactccagga gactaatgcg gcgctgcaag acgtgagaga gctcttgcga
     3061 cagcaggtca aggagatcac cttcctgaag aatacggtga tggaatgtga cgcctgcggt
     3121 ctagacagaa acctgcctcc actagcacct ctgggcccac acccagaaac gctggtgaaa
     3181 gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac
     3241 agcggtaaga tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt
     3301 aaagttctgc tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga gcaactcggt
     3361 cgccgcatac actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat
     3421 cttacggatg gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac
     3481 actgcggcca acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg
     3541 cacaacatgg gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc
     3601 ataccaaacg acgagcgtga caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa
     3661 ctattaactg gcgaactact tactctagct tcccggcaac aattaataga ctggatggag
     3721 gcggataaag ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct
     3781 gataaatctg gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat
     3841 ggtaagccct cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa
     3901 cgaaatagac agatcgctga gataggtgcc tcactgatta agcattgggg ccctgactac
     3961 aaagaccatg acggtgatta taaagatcat gacatcgatt acaaggatga cgatgacaag
     4021 ggccctcacc accaccatca ccactaataa taaggatccc ccgacctcga cctctggcta
     4081 ataaaggaaa tttattttca ttgcaatagt gtgttggaat tttttgtgtc tctcactcgg
     4141 aaggacatat gggagggcaa atcatttggt cgagatccct cggagatctc tagctagagg
     4201 atcgatcccc gccccggacg aactaaacct gactacgaca tctctgcccc ttcttcgcgg
     4261 ggcagtgcat gtaatccctt cagttggttg gtacaacttg ccaactgggc cctgttccac
     4321 atgtgacacg gggggggacc aaacacaaag gggttctctg actgtagttg acatccttat
     4381 aaatggatgt gcacatttgc caacactgag tggctttcat cctggagcag actttgcagt
     4441 ctgtggactg caacacaaca ttgcctttat gtgtaactct tggctgaagc tcttacacca
     4501 atgctggggg acatgtacct cccaggggcc caggaagact acgggaggct acaccaacgt
     4561 caatcagagg ggcctgtgta gctaccgata agcggaccct caagagggca ttagcaatag
     4621 tgtttataag gcccccttgt taaccctaaa cgggtagcat atgcttcccg ggtagtagta
     4681 tatactatcc agactaaccc taattcaata gcatatgtta cccaacggga agcatatgct
     4741 atcgaattag ggttagtaaa agggtcctaa ggaacagcga tatctcccac cccatgagct
     4801 gtcacggttt tatttacatg gggtcaggat tccacgaggg tagtgaacca ttttagtcac
     4861 aagggcagtg gctgaagatc aaggagcggg cagtgaactc tcctgaatct tcgcctgctt
     4921 cttcattctc cttcgtttag ctaatagaat aactgctgag ttgtgaacag taaggtgtat
     4981 gtgaggtgct cgaaaacaag gtttcaggtg acgcccccag aataaaattt ggacgggggg
     5041 ttcagtggtg gcattgtgct atgacaccaa tataaccctc acaaacccct tgggcaataa
     5101 atactagtgt aggaatgaaa cattctgaat atctttaaca atagaaatcc atggggtggg
     5161 gacaagccgt aaagactgga tgtccatctc acacgaattt atggctatgg gcaacacata
     5221 atcctagtgc aatatgatac tggggttatt aagatgtgtc ccaggcaggg accaagacag
     5281 gtgaaccatg ttgttacact ctatttgtaa caaggggaaa gagagtggac gccgacagca
     5341 gcggactcca ctggttgtct ctaacacccc cgaaaattaa acggggctcc acgccaatgg
     5401 ggcccataaa caaagacaag tggccactct tttttttgaa attgtggagt gggggcacgc
     5461 gtcagccccc acacgccgcc ctgcggtttt ggactgtaaa ataagggtgt aataacttgg
     5521 ctgattgtaa ccccgctaac cactgcggtc aaaccacttg cccacaaaac cactaatggc
     5581 accccgggga atacctgcat aagtaggtgg gcgggccaag ataggggcgc gattgctgcg
     5641 atctggagga caaattacac acacttgcgc ctgagcgcca agcacagggt tgttggtcct
     5701 catattcacg aggtcgctga gagcacggtg ggctaatgtt gccatgggta gcatatacta
     5761 cccaaatatc tggatagcat atgctatcct aatctatatc tgggtagcat aggctatcct
     5821 aatctatatc tgggtagcat atgctatcct aatctatatc tgggtagtat atgctatcct
     5881 aatttatatc tgggtagcat aggctatcct aatctatatc tgggtagcat atgctatcct
     5941 aatctatatc tgggtagtat atgctatcct aatctgtatc cgggtagcat atgctatcct
     6001 aatagagatt agggtagtat atgctatcct aatttatatc tgggtagcat atactaccca
     6061 aatatctgga tagcatatgc tatcctaatc tatatctggg tagcatatgc tatcctaatc
     6121 tatatctggg tagcataggc tatcctaatc tatatctggg tagcatatgc tatcctaatc
     6181 tatatctggg tagtatatgc tatcctaatt tatatctggg tagcataggc tatcctaatc
     6241 tatatctggg tagcatatgc tatcctaatc tatatctggg tagtatatgc tatcctaatc
     6301 tgtatccggg tagcatatgc tatcctcatg ataagctgtc aaacatgaga attttcttga
     6361 agacgaaagg gcctcgtgat acgcctattt ttataggtta atgtcatgat aataatggtt
     6421 tcttagacgt caggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt
     6481 ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa
     6541 taatattgaa aaaggaagag tatgagtatt caacatttcc gtgtcgccct tattcccttt
     6601 tttgcggcat tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat
     6661 gctgaagatc agttgggtgc acgagtgggt tacatcgaac tggatctcaa cagcggtaag
     6721 atccttgaga gttttcgccc cgaagaacgt tttccaatga tgagcacttt taaagttctg
     6781 ctatgtggcg cggtattatc ccgtgttgac gccgggcaag agcaactcgg tcgccgcata
     6841 cactattctc agaatgactt ggttgagtac tcaccagtca cagaaaagca tcttacggat
     6901 ggcatgacag taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc
     6961 aacttacttc tgacaacgat cggaggaccg aaggagctaa ccgctttttt gcacaacatg
     7021 ggggatcatg taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac
     7081 gacgagcgtg acaccacgat gcctgcagca atggcaacaa cgttgcgcaa actattaact
     7141 ggcgaactac ttactctagc ttcccggcaa caattaatag actggatgga ggcggataaa
     7201 gttgcaggac cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct
     7261 ggagccggtg agcgtgggtc tcgcggtatc attgcagcac tggggccaga tggtaagccc
     7321 tcccgtatcg tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga
     7381 cagatcgctg agataggtgc ctcactgatt aagcattggt aactgtcaga ccaagtttac
     7441 tcatatatac tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag
     7501 atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg
     7561 tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc
     7621 tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag
     7681 ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtt
     7741 cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac
     7801 ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc
     7861 gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt
     7921 tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt
     7981 gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc
     8041 ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt
     8101 tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca
     8161 ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt
     8221 tgctggcctt ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt
     8281 attaccgcct ttgagtgagc tgataccgct cgccgcagcc gaacgaccga gcgcagcgag
     8341 tcagtgagcg aggaagcgga agagcgccca atacgcaaac cgcctctccc cgcgcgttgg
     8401 ccgattcatt aatgcagctg gcacgacagg tttcccgact ggaaagcggg cagtgagcgc
     8461 aacgcaatta atgtgagtta gctcactcat taggcacccc aggctttaca ctttatgctt
     8521 ccggctcgta tgttgtgtgg aattgtgagc ggataacaat ttcacacagg aaacagctat
     8581 gaccatgatt acgccaagct ctagctagag gtcgaccaat tctcatgttt gacagcttat
     8641 catcgcagat ccgggcaacg ttgttgccat tgctgcaggc gcagaactgg taggtatgga
     8701 agatctatac attgaatcaa tattggcaat tagccatatt agtcattggt tatatagcat
     8761 aaatcaatat tggctattgg ccattgcata cgttgtatct atatcataat atgtacattt
     8821 atattggctc atgtccaata tgaccgccat gttgacattg attattgact agttattaat
     8881 agtaatcaat tacggggtca ttagttcata gcccatatat ggagttccgc gttacataac
     8941 ttacggtaaa tggcccgcct ggctgaccgc ccaacgaccc ccgcccattg acgtcaataa
     9001 tgacgtatgt tcccatagta acgccaatag ggactttcca ttgacgtcaa tgggtggagt
     9061 atttacggta aactgcccac ttggcagtac atcaagtgta tcatatgcca agtccgcccc
     9121 ctattgacgt caatgacggt aaatggcccg cctggcatta tgcccagtac atgaccttac
     9181 gggactttcc tacttggcag tacatctacg tattagtcat cgctattacc atggtgatgc
     9241 ggttttggca gtacaccaat gggcgtggat agcggtttga ctcacgggga tttccaagtc
     9301 tccaccccat tgacgtcaat gggagtttgt tttggcacca aaatcaacgg gactttccaa
     9361 aatgtcgtaa taaccccgcc ccgttgacgc aaatgggcgg taggcgtgta cggtgggagg
     9421 tctatataag cagagctcgt ttagtgaacc gtcagatcct cactctcttc cgcatcgctg
     9481 tctgcgaggg ccagctgttg ggctcgcggt tgaggacaaa ctcttcgcgg tctttccagt
     9541 actcttggat cggaaacccg tcggcctccg aacggtactc cgccaccgag ggacctgagc
     9601 gagtccgcat cgaccggatc ggaaaacct
//
