LOCUS       sequence-208715-        8119 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 8119)
  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 8119)
  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_0526900-COMP-blac-flag-his
FEATURES             Location/Qualifiers
     source          1..8119
                     /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             1651..2436
                     /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             2443..2508
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             2515..2532
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    2570..2625
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(2624..2643)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(4861..4879)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4947..5051
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5052..5120,5121..5912)
                     /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      6083..6671
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6572..6591
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6825..6842
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    6959..6980
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6995..7000,7001..7018,7019..7025)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7033..7049
                     /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     7038..7060
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7057..7073
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7057..7073
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     enhancer        7344..7723
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        7724..7927
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     7878..7898
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     7924..7948
                     /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 ggccgccgcc
      421 tacttcttcg tgaaagaggg cgtggacaag tgcttcgtgg aaagcgtggc caacaacgtg
      481 gtcatcaccg ccacctacga caactacggc ctgaaggacg tgaagtgcct gatcaacatc
      541 aaggaccagc agggcaaggt gctgtactcc cacgacacca gcaagatccg gaagggcaag
      601 atcagctacc tgagcaccaa ggacggcctg taccacatct gcatcagctg ccccagcacc
      661 aactggttca agagcaccgc catcaagtgg tccctgagca tcgaagtggg cggcagcgac
      721 atcgaccccg agaacctggc caagaagtcc gagctgagcg agacactgac catcctgaac
      781 aacctgaaga aaaagttcaa cagcatgaag ctgcagcaga tctaccagaa acagatggcc
      841 agcaacctgt acgagtacaa caaggccgtg cacaacggcg cgccgtcgac ctccatcacg
      901 gcctataaga gtgaggggga gtcagcggag ttctccttcc cactcaacct tggagaggaa
      961 agcctgcagg gagagttgag atggaaggca gagaaggctc cttcttccca gtcctggatc
     1021 accttctccc taaagaacca aaaggtgtct gtgcagaagt ctactagcaa ccccaagttc
     1081 cagctgtccg aaacgctccc actcaccctt cagatacccc aggtctccct tcagtttgct
     1141 ggttctggca acctgaccct gactctggac agagggatac tgtatcagga agtgaacctg
     1201 gtggtgatga aagtgactca gcccgacagc aacactttga cctgtgaggt gatgggaccc
     1261 acctcaccca agatgagact gatcttgaag caggagaatc aggaggccag ggtctccagg
     1321 caggagaaag tgattcaagt gcaggcccct gaagcagggg tgtggcaatg tctactgagt
     1381 gaaggtgaag aggtcaagat ggactccaag atccaggttt tatccaaagg gttgaattcc
     1441 ggcggcggct ccggaggcgg taccggtggc ggcgacctag ccccacagat gcttcgagaa
     1501 ctccaggaga ctaatgcggc gctgcaagac gtgagagagc tcttgcgaca gcaggtcaag
     1561 gagatcacct tcctgaagaa tacggtgatg gaatgtgacg cctgcggtct agacagaaac
     1621 ctgcctccac tagcacctct gggcccacac ccagaaacgc tggtgaaagt aaaagatgct
     1681 gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag cggtaagatc
     1741 cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa agttctgcta
     1801 tgtggcgcgg tattatcccg tattgacgcc gggcaagagc aactcggtcg ccgcatacac
     1861 tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct tacggatggc
     1921 atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac
     1981 ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca caacatgggg
     2041 gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat accaaacgac
     2101 gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact attaactggc
     2161 gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc ggataaagtt
     2221 gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga taaatctgga
     2281 gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg taagccctcc
     2341 cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag
     2401 atcgctgaga taggtgcctc actgattaag cattggggcc ctgactacaa agaccatgac
     2461 ggtgattata aagatcatga catcgattac aaggatgacg atgacaaggg ccctcaccac
     2521 caccatcacc actaattaat taaggatccc ccgacctcga cctctggcta ataaaggaaa
     2581 tttattttca ttgcaatagt gtgttggaat tttttgtgtc tctcactcgg aaggacatat
     2641 gggagggcaa atcatttggt cgagatccct cggagatctc tagctagagg atcgatcccc
     2701 gccccggacg aactaaacct gactacgaca tctctgcccc ttcttcgcgg ggcagtgcat
     2761 gtaatccctt cagttggttg gtacaacttg ccaactgggc cctgttccac atgtgacacg
     2821 gggggggacc aaacacaaag gggttctctg actgtagttg acatccttat aaatggatgt
     2881 gcacatttgc caacactgag tggctttcat cctggagcag actttgcagt ctgtggactg
     2941 caacacaaca ttgcctttat gtgtaactct tggctgaagc tcttacacca atgctggggg
     3001 acatgtacct cccaggggcc caggaagact acgggaggct acaccaacgt caatcagagg
     3061 ggcctgtgta gctaccgata agcggaccct caagagggca ttagcaatag tgtttataag
     3121 gcccccttgt taaccctaaa cgggtagcat atgcttcccg ggtagtagta tatactatcc
     3181 agactaaccc taattcaata gcatatgtta cccaacggga agcatatgct atcgaattag
     3241 ggttagtaaa agggtcctaa ggaacagcga tatctcccac cccatgagct gtcacggttt
     3301 tatttacatg gggtcaggat tccacgaggg tagtgaacca ttttagtcac aagggcagtg
     3361 gctgaagatc aaggagcggg cagtgaactc tcctgaatct tcgcctgctt cttcattctc
     3421 cttcgtttag ctaatagaat aactgctgag ttgtgaacag taaggtgtat gtgaggtgct
     3481 cgaaaacaag gtttcaggtg acgcccccag aataaaattt ggacgggggg ttcagtggtg
     3541 gcattgtgct atgacaccaa tataaccctc acaaacccct tgggcaataa atactagtgt
     3601 aggaatgaaa cattctgaat atctttaaca atagaaatcc atggggtggg gacaagccgt
     3661 aaagactgga tgtccatctc acacgaattt atggctatgg gcaacacata atcctagtgc
     3721 aatatgatac tggggttatt aagatgtgtc ccaggcaggg accaagacag gtgaaccatg
     3781 ttgttacact ctatttgtaa caaggggaaa gagagtggac gccgacagca gcggactcca
     3841 ctggttgtct ctaacacccc cgaaaattaa acggggctcc acgccaatgg ggcccataaa
     3901 caaagacaag tggccactct tttttttgaa attgtggagt gggggcacgc gtcagccccc
     3961 acacgccgcc ctgcggtttt ggactgtaaa ataagggtgt aataacttgg ctgattgtaa
     4021 ccccgctaac cactgcggtc aaaccacttg cccacaaaac cactaatggc accccgggga
     4081 atacctgcat aagtaggtgg gcgggccaag ataggggcgc gattgctgcg atctggagga
     4141 caaattacac acacttgcgc ctgagcgcca agcacagggt tgttggtcct catattcacg
     4201 aggtcgctga gagcacggtg ggctaatgtt gccatgggta gcatatacta cccaaatatc
     4261 tggatagcat atgctatcct aatctatatc tgggtagcat aggctatcct aatctatatc
     4321 tgggtagcat atgctatcct aatctatatc tgggtagtat atgctatcct aatttatatc
     4381 tgggtagcat aggctatcct aatctatatc tgggtagcat atgctatcct aatctatatc
     4441 tgggtagtat atgctatcct aatctgtatc cgggtagcat atgctatcct aatagagatt
     4501 agggtagtat atgctatcct aatttatatc tgggtagcat atactaccca aatatctgga
     4561 tagcatatgc tatcctaatc tatatctggg tagcatatgc tatcctaatc tatatctggg
     4621 tagcataggc tatcctaatc tatatctggg tagcatatgc tatcctaatc tatatctggg
     4681 tagtatatgc tatcctaatt tatatctggg tagcataggc tatcctaatc tatatctggg
     4741 tagcatatgc tatcctaatc tatatctggg tagtatatgc tatcctaatc tgtatccggg
     4801 tagcatatgc tatcctcatg ataagctgtc aaacatgaga attttcttga agacgaaagg
     4861 gcctcgtgat acgcctattt ttataggtta atgtcatgat aataatggtt tcttagacgt
     4921 caggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac
     4981 attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa
     5041 aaaggaagag tatgagtatt caacatttcc gtgtcgccct tattcccttt tttgcggcat
     5101 tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat gctgaagatc
     5161 agttgggtgc acgagtgggt tacatcgaac tggatctcaa cagcggtaag atccttgaga
     5221 gttttcgccc cgaagaacgt tttccaatga tgagcacttt taaagttctg ctatgtggcg
     5281 cggtattatc ccgtgttgac gccgggcaag agcaactcgg tcgccgcata cactattctc
     5341 agaatgactt ggttgagtac tcaccagtca cagaaaagca tcttacggat ggcatgacag
     5401 taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc aacttacttc
     5461 tgacaacgat cggaggaccg aaggagctaa ccgctttttt gcacaacatg ggggatcatg
     5521 taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac gacgagcgtg
     5581 acaccacgat gcctgcagca atggcaacaa cgttgcgcaa actattaact ggcgaactac
     5641 ttactctagc ttcccggcaa caattaatag actggatgga ggcggataaa gttgcaggac
     5701 cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct ggagccggtg
     5761 agcgtgggtc tcgcggtatc attgcagcac tggggccaga tggtaagccc tcccgtatcg
     5821 tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga cagatcgctg
     5881 agataggtgc ctcactgatt aagcattggt aactgtcaga ccaagtttac tcatatatac
     5941 tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg
     6001 ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg
     6061 tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc
     6121 aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc
     6181 tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt
     6241 agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc
     6301 taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact
     6361 caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac
     6421 agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag
     6481 aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg
     6541 gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg
     6601 tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga
     6661 gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt
     6721 ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt attaccgcct
     6781 ttgagtgagc tgataccgct cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg
     6841 aggaagcgga agagcgccca atacgcaaac cgcctctccc cgcgcgttgg ccgattcatt
     6901 aatgcagctg gcacgacagg tttcccgact ggaaagcggg cagtgagcgc aacgcaatta
     6961 atgtgagtta gctcactcat taggcacccc aggctttaca ctttatgctt ccggctcgta
     7021 tgttgtgtgg aattgtgagc ggataacaat ttcacacagg aaacagctat gaccatgatt
     7081 acgccaagct ctagctagag gtcgaccaat tctcatgttt gacagcttat catcgcagat
     7141 ccgggcaacg ttgttgccat tgctgcaggc gcagaactgg taggtatgga agatctatac
     7201 attgaatcaa tattggcaat tagccatatt agtcattggt tatatagcat aaatcaatat
     7261 tggctattgg ccattgcata cgttgtatct atatcataat atgtacattt atattggctc
     7321 atgtccaata tgaccgccat gttgacattg attattgact agttattaat agtaatcaat
     7381 tacggggtca ttagttcata gcccatatat ggagttccgc gttacataac ttacggtaaa
     7441 tggcccgcct ggctgaccgc ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt
     7501 tcccatagta acgccaatag ggactttcca ttgacgtcaa tgggtggagt atttacggta
     7561 aactgcccac ttggcagtac atcaagtgta tcatatgcca agtccgcccc ctattgacgt
     7621 caatgacggt aaatggcccg cctggcatta tgcccagtac atgaccttac gggactttcc
     7681 tacttggcag tacatctacg tattagtcat cgctattacc atggtgatgc ggttttggca
     7741 gtacaccaat gggcgtggat agcggtttga ctcacgggga tttccaagtc tccaccccat
     7801 tgacgtcaat gggagtttgt tttggcacca aaatcaacgg gactttccaa aatgtcgtaa
     7861 taaccccgcc ccgttgacgc aaatgggcgg taggcgtgta cggtgggagg tctatataag
     7921 cagagctcgt ttagtgaacc gtcagatcct cactctcttc cgcatcgctg tctgcgaggg
     7981 ccagctgttg ggctcgcggt tgaggacaaa ctcttcgcgg tctttccagt actcttggat
     8041 cggaaacccg tcggcctccg aacggtactc cgccaccgag ggacctgagc gagtccgcat
     8101 cgaccggatc ggaaaacct
//
