LOCUS       sequence-208747-        9043 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 9043)
  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 9043)
  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_0620000-COMP-blac-flag-his
FEATURES             Location/Qualifiers
     source          1..9043
                     /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             2575..3360
                     /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             3367..3432
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             3439..3456
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    3494..3549
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(3548..3567)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(5785..5803)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5871..5975
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5976..6044,6045..6836)
                     /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      7007..7595
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7496..7515
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7749..7766
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    7883..7904
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7919..7924,7925..7942,7943..7949)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7957..7973
                     /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     7962..7984
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7981..7997
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7981..7997
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     enhancer        8268..8647
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        8648..8851
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     8802..8822
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     8848..8872
                     /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 ggccgccaag
      421 gacgatggac agagacacgg cgaggacagc gacagcatga acaagtacaa ctacatctac
      481 cggcccaacg ccgacaacgg catccacaac aacctgcggc ccagcagcta catcgacatg
      541 gaacacattg ccaagggccg ggacaacaca gccggcctga gagagaacga gctgttcgac
      601 atccagaacg acgagaacag caacatgggc cccgaccagg aaaaagtcat caccctgagc
      661 gacaagatct acatcgataa gatcaagaac atcatcgagt accggaagtg gcgggccaag
      721 agcaagtaca gaagccccgc caagcagccc gaagaggaca tgttcgtgga aggctacatc
      781 cccatcaagc ggcgggacga caacctgctg cctcacgaga aggccgagaa ggacttcgag
      841 gaaatcatcc agaaatacgt ggacctcgag aacaagctgc agaagcagaa agagctggaa
      901 gaggccgagc ggcggaagaa gaagctggac gacgagcaga agcggcagaa ggatctcgag
      961 gacaccaacc ggaaagagaa tgacgaggaa cagagctaca agaagctcga ggacctggaa
     1021 ctggaaaaca tcgacaccaa gatcaacaac aacgacgcct acgacgtgga aaacggcttc
     1081 aacgacgaca acaagctgta cacaaagatt aacaagatcg tcagccaggt gtcccagaac
     1141 aatgagctgt acgtgaacat catgaagtac atcacactgg tctacaccag ccacgtggac
     1201 atcaagcagg acgacttcac caacggcgcc atcagcatct ttctgacctt cgagaacccc
     1261 aagatgggca acgacctggc caatatcaat atcgccttct tcgccagcga gcagcggacc
     1321 ggcaccaacg aggtggccga ggccagaagc aactacctga ccaactacaa catgaatgag
     1381 ctgctcgagg ataacgccgc tggccggctg ctgagccagg acgagtacat caaagaactc
     1441 gtgaagtaca accacgccat ctccagcagc aacaaagaga acctgaagaa cgagaactac
     1501 gacgtcaaca tgaacattgc taagatcctg cagtatatca tcgacaacga catcaacctg
     1561 gtggacaact tcatcacctt caacagagcc aacgagcagg aagtgaccat cagcaagctg
     1621 gatgactttc tgcacgagtg ctccaaagtg aagcccgagg atatcaagaa ggatgagatc
     1681 cagaagatca tcaagaccat caacaagaaa gagcagatta tcaaagaagt gaagaacagc
     1741 ctgatcaaga aagacatcga gaagtgcaag ctctacacca ccatcctgat gttcggcagc
     1801 ggcgcgccgt cgacctccat cacggcctat aagagtgagg gggagtcagc ggagttctcc
     1861 ttcccactca accttggaga ggaaagcctg cagggagagt tgagatggaa ggcagagaag
     1921 gctccttctt cccagtcctg gatcaccttc tccctaaaga accaaaaggt gtctgtgcag
     1981 aagtctacta gcaaccccaa gttccagctg tccgaaacgc tcccactcac ccttcagata
     2041 ccccaggtct cccttcagtt tgctggttct ggcaacctga ccctgactct ggacagaggg
     2101 atactgtatc aggaagtgaa cctggtggtg atgaaagtga ctcagcccga cagcaacact
     2161 ttgacctgtg aggtgatggg acccacctca cccaagatga gactgatctt gaagcaggag
     2221 aatcaggagg ccagggtctc caggcaggag aaagtgattc aagtgcaggc ccctgaagca
     2281 ggggtgtggc aatgtctact gagtgaaggt gaagaggtca agatggactc caagatccag
     2341 gttttatcca aagggttgaa ttccggcggc ggctccggag gcggtaccgg tggcggcgac
     2401 ctagccccac agatgcttcg agaactccag gagactaatg cggcgctgca agacgtgaga
     2461 gagctcttgc gacagcaggt caaggagatc accttcctga agaatacggt gatggaatgt
     2521 gacgcctgcg gtctagacag aaacctgcct ccactagcac ctctgggccc acacccagaa
     2581 acgctggtga aagtaaaaga tgctgaagat cagttgggtg cacgagtggg ttacatcgaa
     2641 ctggatctca acagcggtaa gatccttgag agttttcgcc ccgaagaacg ttttccaatg
     2701 atgagcactt ttaaagttct gctatgtggc gcggtattat cccgtattga cgccgggcaa
     2761 gagcaactcg gtcgccgcat acactattct cagaatgact tggttgagta ctcaccagtc
     2821 acagaaaagc atcttacgga tggcatgaca gtaagagaat tatgcagtgc tgccataacc
     2881 atgagtgata acactgcggc caacttactt ctgacaacga tcggaggacc gaaggagcta
     2941 accgcttttt tgcacaacat gggggatcat gtaactcgcc ttgatcgttg ggaaccggag
     3001 ctgaatgaag ccataccaaa cgacgagcgt gacaccacga tgcctgtagc aatggcaaca
     3061 acgttgcgca aactattaac tggcgaacta cttactctag cttcccggca acaattaata
     3121 gactggatgg aggcggataa agttgcagga ccacttctgc gctcggccct tccggctggc
     3181 tggtttattg ctgataaatc tggagccggt gagcgtgggt ctcgcggtat cattgcagca
     3241 ctggggccag atggtaagcc ctcccgtatc gtagttatct acacgacggg gagtcaggca
     3301 actatggatg aacgaaatag acagatcgct gagataggtg cctcactgat taagcattgg
     3361 ggccctgact acaaagacca tgacggtgat tataaagatc atgacatcga ttacaaggat
     3421 gacgatgaca agggccctca ccaccaccat caccactaat taattaagga tcccccgacc
     3481 tcgacctctg gctaataaag gaaatttatt ttcattgcaa tagtgtgttg gaattttttg
     3541 tgtctctcac tcggaaggac atatgggagg gcaaatcatt tggtcgagat ccctcggaga
     3601 tctctagcta gaggatcgat ccccgccccg gacgaactaa acctgactac gacatctctg
     3661 ccccttcttc gcggggcagt gcatgtaatc ccttcagttg gttggtacaa cttgccaact
     3721 gggccctgtt ccacatgtga cacggggggg gaccaaacac aaaggggttc tctgactgta
     3781 gttgacatcc ttataaatgg atgtgcacat ttgccaacac tgagtggctt tcatcctgga
     3841 gcagactttg cagtctgtgg actgcaacac aacattgcct ttatgtgtaa ctcttggctg
     3901 aagctcttac accaatgctg ggggacatgt acctcccagg ggcccaggaa gactacggga
     3961 ggctacacca acgtcaatca gaggggcctg tgtagctacc gataagcgga ccctcaagag
     4021 ggcattagca atagtgttta taaggccccc ttgttaaccc taaacgggta gcatatgctt
     4081 cccgggtagt agtatatact atccagacta accctaattc aatagcatat gttacccaac
     4141 gggaagcata tgctatcgaa ttagggttag taaaagggtc ctaaggaaca gcgatatctc
     4201 ccaccccatg agctgtcacg gttttattta catggggtca ggattccacg agggtagtga
     4261 accattttag tcacaagggc agtggctgaa gatcaaggag cgggcagtga actctcctga
     4321 atcttcgcct gcttcttcat tctccttcgt ttagctaata gaataactgc tgagttgtga
     4381 acagtaaggt gtatgtgagg tgctcgaaaa caaggtttca ggtgacgccc ccagaataaa
     4441 atttggacgg ggggttcagt ggtggcattg tgctatgaca ccaatataac cctcacaaac
     4501 cccttgggca ataaatacta gtgtaggaat gaaacattct gaatatcttt aacaatagaa
     4561 atccatgggg tggggacaag ccgtaaagac tggatgtcca tctcacacga atttatggct
     4621 atgggcaaca cataatccta gtgcaatatg atactggggt tattaagatg tgtcccaggc
     4681 agggaccaag acaggtgaac catgttgtta cactctattt gtaacaaggg gaaagagagt
     4741 ggacgccgac agcagcggac tccactggtt gtctctaaca cccccgaaaa ttaaacgggg
     4801 ctccacgcca atggggccca taaacaaaga caagtggcca ctcttttttt tgaaattgtg
     4861 gagtgggggc acgcgtcagc ccccacacgc cgccctgcgg ttttggactg taaaataagg
     4921 gtgtaataac ttggctgatt gtaaccccgc taaccactgc ggtcaaacca cttgcccaca
     4981 aaaccactaa tggcaccccg gggaatacct gcataagtag gtgggcgggc caagataggg
     5041 gcgcgattgc tgcgatctgg aggacaaatt acacacactt gcgcctgagc gccaagcaca
     5101 gggttgttgg tcctcatatt cacgaggtcg ctgagagcac ggtgggctaa tgttgccatg
     5161 ggtagcatat actacccaaa tatctggata gcatatgcta tcctaatcta tatctgggta
     5221 gcataggcta tcctaatcta tatctgggta gcatatgcta tcctaatcta tatctgggta
     5281 gtatatgcta tcctaattta tatctgggta gcataggcta tcctaatcta tatctgggta
     5341 gcatatgcta tcctaatcta tatctgggta gtatatgcta tcctaatctg tatccgggta
     5401 gcatatgcta tcctaataga gattagggta gtatatgcta tcctaattta tatctgggta
     5461 gcatatacta cccaaatatc tggatagcat atgctatcct aatctatatc tgggtagcat
     5521 atgctatcct aatctatatc tgggtagcat aggctatcct aatctatatc tgggtagcat
     5581 atgctatcct aatctatatc tgggtagtat atgctatcct aatttatatc tgggtagcat
     5641 aggctatcct aatctatatc tgggtagcat atgctatcct aatctatatc tgggtagtat
     5701 atgctatcct aatctgtatc cgggtagcat atgctatcct catgataagc tgtcaaacat
     5761 gagaattttc ttgaagacga aagggcctcg tgatacgcct atttttatag gttaatgtca
     5821 tgataataat ggtttcttag acgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc
     5881 ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct
     5941 gataaatgct tcaataatat tgaaaaagga agagtatgag tattcaacat ttccgtgtcg
     6001 cccttattcc cttttttgcg gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg
     6061 tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt gggttacatc gaactggatc
     6121 tcaacagcgg taagatcctt gagagttttc gccccgaaga acgttttcca atgatgagca
     6181 cttttaaagt tctgctatgt ggcgcggtat tatcccgtgt tgacgccggg caagagcaac
     6241 tcggtcgccg catacactat tctcagaatg acttggttga gtactcacca gtcacagaaa
     6301 agcatcttac ggatggcatg acagtaagag aattatgcag tgctgccata accatgagtg
     6361 ataacactgc ggccaactta cttctgacaa cgatcggagg accgaaggag ctaaccgctt
     6421 ttttgcacaa catgggggat catgtaactc gccttgatcg ttgggaaccg gagctgaatg
     6481 aagccatacc aaacgacgag cgtgacacca cgatgcctgc agcaatggca acaacgttgc
     6541 gcaaactatt aactggcgaa ctacttactc tagcttcccg gcaacaatta atagactgga
     6601 tggaggcgga taaagttgca ggaccacttc tgcgctcggc ccttccggct ggctggttta
     6661 ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc
     6721 cagatggtaa gccctcccgt atcgtagtta tctacacgac ggggagtcag gcaactatgg
     6781 atgaacgaaa tagacagatc gctgagatag gtgcctcact gattaagcat tggtaactgt
     6841 cagaccaagt ttactcatat atactttaga ttgatttaaa acttcatttt taatttaaaa
     6901 ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt
     6961 cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt
     7021 ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt
     7081 tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga
     7141 taccaaatac tgttcttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag
     7201 caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata
     7261 agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg
     7321 gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga
     7381 gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca
     7441 ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa
     7501 acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt
     7561 tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac
     7621 ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta tcccctgatt
     7681 ctgtggataa ccgtattacc gcctttgagt gagctgatac cgctcgccgc agccgaacga
     7741 ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg cccaatacgc aaaccgcctc
     7801 tccccgcgcg ttggccgatt cattaatgca gctggcacga caggtttccc gactggaaag
     7861 cgggcagtga gcgcaacgca attaatgtga gttagctcac tcattaggca ccccaggctt
     7921 tacactttat gcttccggct cgtatgttgt gtggaattgt gagcggataa caatttcaca
     7981 caggaaacag ctatgaccat gattacgcca agctctagct agaggtcgac caattctcat
     8041 gtttgacagc ttatcatcgc agatccgggc aacgttgttg ccattgctgc aggcgcagaa
     8101 ctggtaggta tggaagatct atacattgaa tcaatattgg caattagcca tattagtcat
     8161 tggttatata gcataaatca atattggcta ttggccattg catacgttgt atctatatca
     8221 taatatgtac atttatattg gctcatgtcc aatatgaccg ccatgttgac attgattatt
     8281 gactagttat taatagtaat caattacggg gtcattagtt catagcccat atatggagtt
     8341 ccgcgttaca taacttacgg taaatggccc gcctggctga ccgcccaacg acccccgccc
     8401 attgacgtca ataatgacgt atgttcccat agtaacgcca atagggactt tccattgacg
     8461 tcaatgggtg gagtatttac ggtaaactgc ccacttggca gtacatcaag tgtatcatat
     8521 gccaagtccg ccccctattg acgtcaatga cggtaaatgg cccgcctggc attatgccca
     8581 gtacatgacc ttacgggact ttcctacttg gcagtacatc tacgtattag tcatcgctat
     8641 taccatggtg atgcggtttt ggcagtacac caatgggcgt ggatagcggt ttgactcacg
     8701 gggatttcca agtctccacc ccattgacgt caatgggagt ttgttttggc accaaaatca
     8761 acgggacttt ccaaaatgtc gtaataaccc cgccccgttg acgcaaatgg gcggtaggcg
     8821 tgtacggtgg gaggtctata taagcagagc tcgtttagtg aaccgtcaga tcctcactct
     8881 cttccgcatc gctgtctgcg agggccagct gttgggctcg cggttgagga caaactcttc
     8941 gcggtctttc cagtactctt ggatcggaaa cccgtcggcc tccgaacggt actccgccac
     9001 cgagggacct gagcgagtcc gcatcgaccg gatcggaaaa cct
//
