LOCUS       sequence-224478-        9140 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 9140)
  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 9140)
  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_0505700-COMP-blac-flag-his
FEATURES             Location/Qualifiers
     source          1..9140
                     /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             2674..3459
                     /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             3466..3531
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             3538..3555
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    3591..3646
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(3645..3664)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(5882..5900)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5968..6072
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6073..6141,6142..6933)
                     /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      7104..7692
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7593..7612
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7846..7863
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    7980..8001
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(8016..8021,8022..8039,8040..8046)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8054..8070
                     /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     8059..8081
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     8078..8094
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     8078..8094
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     enhancer        8365..8744
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        8745..8948
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     8899..8919
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     8945..8969
                     /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 ggccgccgaa
      421 gagtacaaga tcaacaaaga ggaagagaac ggcaagcaca acaaagaaga gatcaagaac
      481 gagaagtacg acaacaacaa ctacgagaac aagaacaatg agaataacaa gaatatcgac
      541 agcaacaacc gcgaggacca catcatgaag ctgctgccca tctacaagga caaggataac
      601 gacatcatcg acgagaacat caacaacaag attaactact atgagaagga caacatcaag
      661 caggaaatca agcacaatac cgagaacgat aacaataata acaacaacat cttcatccag
      721 gaaaaggaca atatccagct gtacgtggac aagaaagagc acctggacga gcggaaccag
      781 aacgtgatcc acgagaatat gaagcaggaa caggaccgga acaaagagac acagaagggc
      841 aacgatatcg acgagtccaa gaagcagaaa gagaaagaac agcacaccaa ccagctgatc
      901 gacaaccgga acgaccagat cagcaacgtg gaaaaagagg acatccacaa cggctacgag
      961 aatgacaagg acaccgagca gaaggatgac aacaaggaag ataagaagga tatcgagcag
     1021 gtcgacgaca ataaggagga caaaaaggac attgaacagg tcgacgataa caaagaggat
     1081 aagaaagaca tcgaacaggt ggacgacaac aaagaagaca aaaaagatat tgagcaggtc
     1141 gatgacaaca aagaagataa gaaagacaca gagcaggtcg acgataacaa agaggacaaa
     1201 aaggatacag aacaggtgga cgataataag gaagataaga agtacattga gcagattgat
     1261 aagaacaagg aggacaagaa ggacaccgag caggtcgacg acaacaaaga ggataagaag
     1321 gaccctcacc agaccgacga caagaacgac ggcgccagct gcgaggaaga aaagaaagtc
     1381 atcatgagcg agattaagac catcaaggag gatctggaca actgcaatat cgccaacaag
     1441 aaagacaaga atgagtgcat cgtgaaagtg aacgaggccg agcggaagct gaacctgtgc
     1501 aaagaaaagg ccctgaagtg caagggccag atcaagggca tccaccacaa gtgcgacgag
     1561 agaatcgacc ggaagaacaa agaaatgatc atcctgcagg gcatcatcaa caagctggaa
     1621 aacaagatcg acgaggataa aaaaaatgat aacaaaaact tccaggatca catcaagaag
     1681 tccgagggca agaagaacga aatccccgtc gaaaacaaaa acaccaagga caacgtgaag
     1741 cgcaacgaga agaatgtgca ccacgaggac tacagcagcc accacaacat ctaccggcag
     1801 gaggataact acctgatcag ctacgagatc ctgaagaact acttcgagaa gatctttgag
     1861 atctacaaga ccctgtacat catcatcgtg gaaaagaccg gcgcgccgtc gacctccatc
     1921 acggcctata agagtgaggg ggagtcagcg gagttctcct tcccactcaa ccttggagag
     1981 gaaagcctgc agggagagtt gagatggaag gcagagaagg ctccttcttc ccagtcctgg
     2041 atcaccttct ccctaaagaa ccaaaaggtg tctgtgcaga agtctactag caaccccaag
     2101 ttccagctgt ccgaaacgct cccactcacc cttcagatac cccaggtctc ccttcagttt
     2161 gctggttctg gcaacctgac cctgactctg gacagaggga tactgtatca ggaagtgaac
     2221 ctggtggtga tgaaagtgac tcagcccgac agcaacactt tgacctgtga ggtgatggga
     2281 cccacctcac ccaagatgag actgatcttg aagcaggaga atcaggaggc cagggtctcc
     2341 aggcaggaga aagtgattca agtgcaggcc cctgaagcag gggtgtggca atgtctactg
     2401 agtgaaggtg aagaggtcaa gatggactcc aagatccagg ttttatccaa agggttgaat
     2461 tccggcggcg gctccggagg cggtaccggt ggcggcgacc tagccccaca gatgcttcga
     2521 gaactccagg agactaatgc ggcgctgcaa gacgtgagag agctcttgcg acagcaggtc
     2581 aaggagatca ccttcctgaa gaatacggtg atggaatgtg acgcctgcgg tctagacaga
     2641 aacctgcctc cactagcacc tctgggccca cacccagaaa cgctggtgaa agtaaaagat
     2701 gctgaagatc agttgggtgc acgagtgggt tacatcgaac tggatctcaa cagcggtaag
     2761 atccttgaga gttttcgccc cgaagaacgt tttccaatga tgagcacttt taaagttctg
     2821 ctatgtggcg cggtattatc ccgtattgac gccgggcaag agcaactcgg tcgccgcata
     2881 cactattctc agaatgactt ggttgagtac tcaccagtca cagaaaagca tcttacggat
     2941 ggcatgacag taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc
     3001 aacttacttc tgacaacgat cggaggaccg aaggagctaa ccgctttttt gcacaacatg
     3061 ggggatcatg taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac
     3121 gacgagcgtg acaccacgat gcctgtagca atggcaacaa cgttgcgcaa actattaact
     3181 ggcgaactac ttactctagc ttcccggcaa caattaatag actggatgga ggcggataaa
     3241 gttgcaggac cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct
     3301 ggagccggtg agcgtgggtc tcgcggtatc attgcagcac tggggccaga tggtaagccc
     3361 tcccgtatcg tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga
     3421 cagatcgctg agataggtgc ctcactgatt aagcattggg gccctgacta caaagaccat
     3481 gacggtgatt ataaagatca tgacatcgat tacaaggatg acgatgacaa gggccctcac
     3541 caccaccatc accactaata ataaggatcc cccgacctcg acctctggct aataaaggaa
     3601 atttattttc attgcaatag tgtgttggaa ttttttgtgt ctctcactcg gaaggacata
     3661 tgggagggca aatcatttgg tcgagatccc tcggagatct ctagctagag gatcgatccc
     3721 cgccccggac gaactaaacc tgactacgac atctctgccc cttcttcgcg gggcagtgca
     3781 tgtaatccct tcagttggtt ggtacaactt gccaactggg ccctgttcca catgtgacac
     3841 ggggggggac caaacacaaa ggggttctct gactgtagtt gacatcctta taaatggatg
     3901 tgcacatttg ccaacactga gtggctttca tcctggagca gactttgcag tctgtggact
     3961 gcaacacaac attgccttta tgtgtaactc ttggctgaag ctcttacacc aatgctgggg
     4021 gacatgtacc tcccaggggc ccaggaagac tacgggaggc tacaccaacg tcaatcagag
     4081 gggcctgtgt agctaccgat aagcggaccc tcaagagggc attagcaata gtgtttataa
     4141 ggcccccttg ttaaccctaa acgggtagca tatgcttccc gggtagtagt atatactatc
     4201 cagactaacc ctaattcaat agcatatgtt acccaacggg aagcatatgc tatcgaatta
     4261 gggttagtaa aagggtccta aggaacagcg atatctccca ccccatgagc tgtcacggtt
     4321 ttatttacat ggggtcagga ttccacgagg gtagtgaacc attttagtca caagggcagt
     4381 ggctgaagat caaggagcgg gcagtgaact ctcctgaatc ttcgcctgct tcttcattct
     4441 ccttcgttta gctaatagaa taactgctga gttgtgaaca gtaaggtgta tgtgaggtgc
     4501 tcgaaaacaa ggtttcaggt gacgccccca gaataaaatt tggacggggg gttcagtggt
     4561 ggcattgtgc tatgacacca atataaccct cacaaacccc ttgggcaata aatactagtg
     4621 taggaatgaa acattctgaa tatctttaac aatagaaatc catggggtgg ggacaagccg
     4681 taaagactgg atgtccatct cacacgaatt tatggctatg ggcaacacat aatcctagtg
     4741 caatatgata ctggggttat taagatgtgt cccaggcagg gaccaagaca ggtgaaccat
     4801 gttgttacac tctatttgta acaaggggaa agagagtgga cgccgacagc agcggactcc
     4861 actggttgtc tctaacaccc ccgaaaatta aacggggctc cacgccaatg gggcccataa
     4921 acaaagacaa gtggccactc ttttttttga aattgtggag tgggggcacg cgtcagcccc
     4981 cacacgccgc cctgcggttt tggactgtaa aataagggtg taataacttg gctgattgta
     5041 accccgctaa ccactgcggt caaaccactt gcccacaaaa ccactaatgg caccccgggg
     5101 aatacctgca taagtaggtg ggcgggccaa gataggggcg cgattgctgc gatctggagg
     5161 acaaattaca cacacttgcg cctgagcgcc aagcacaggg ttgttggtcc tcatattcac
     5221 gaggtcgctg agagcacggt gggctaatgt tgccatgggt agcatatact acccaaatat
     5281 ctggatagca tatgctatcc taatctatat ctgggtagca taggctatcc taatctatat
     5341 ctgggtagca tatgctatcc taatctatat ctgggtagta tatgctatcc taatttatat
     5401 ctgggtagca taggctatcc taatctatat ctgggtagca tatgctatcc taatctatat
     5461 ctgggtagta tatgctatcc taatctgtat ccgggtagca tatgctatcc taatagagat
     5521 tagggtagta tatgctatcc taatttatat ctgggtagca tatactaccc aaatatctgg
     5581 atagcatatg ctatcctaat ctatatctgg gtagcatatg ctatcctaat ctatatctgg
     5641 gtagcatagg ctatcctaat ctatatctgg gtagcatatg ctatcctaat ctatatctgg
     5701 gtagtatatg ctatcctaat ttatatctgg gtagcatagg ctatcctaat ctatatctgg
     5761 gtagcatatg ctatcctaat ctatatctgg gtagtatatg ctatcctaat ctgtatccgg
     5821 gtagcatatg ctatcctcat gataagctgt caaacatgag aattttcttg aagacgaaag
     5881 ggcctcgtga tacgcctatt tttataggtt aatgtcatga taataatggt ttcttagacg
     5941 tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata
     6001 cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga
     6061 aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc ttattccctt ttttgcggca
     6121 ttttgccttc ctgtttttgc tcacccagaa acgctggtga aagtaaaaga tgctgaagat
     6181 cagttgggtg cacgagtggg ttacatcgaa ctggatctca acagcggtaa gatccttgag
     6241 agttttcgcc ccgaagaacg ttttccaatg atgagcactt ttaaagttct gctatgtggc
     6301 gcggtattat cccgtgttga cgccgggcaa gagcaactcg gtcgccgcat acactattct
     6361 cagaatgact tggttgagta ctcaccagtc acagaaaagc atcttacgga tggcatgaca
     6421 gtaagagaat tatgcagtgc tgccataacc atgagtgata acactgcggc caacttactt
     6481 ctgacaacga tcggaggacc gaaggagcta accgcttttt tgcacaacat gggggatcat
     6541 gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt
     6601 gacaccacga tgcctgcagc aatggcaaca acgttgcgca aactattaac tggcgaacta
     6661 cttactctag cttcccggca acaattaata gactggatgg aggcggataa agttgcagga
     6721 ccacttctgc gctcggccct tccggctggc tggtttattg ctgataaatc tggagccggt
     6781 gagcgtgggt ctcgcggtat cattgcagca ctggggccag atggtaagcc ctcccgtatc
     6841 gtagttatct acacgacggg gagtcaggca actatggatg aacgaaatag acagatcgct
     6901 gagataggtg cctcactgat taagcattgg taactgtcag accaagttta ctcatatata
     6961 ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt
     7021 gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc
     7081 gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg
     7141 caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact
     7201 ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt tcttctagtg
     7261 tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg
     7321 ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac
     7381 tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca
     7441 cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga
     7501 gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc
     7561 ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct
     7621 gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg
     7681 agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct
     7741 tttgctcaca tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc
     7801 tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc
     7861 gaggaagcgg aagagcgccc aatacgcaaa ccgcctctcc ccgcgcgttg gccgattcat
     7921 taatgcagct ggcacgacag gtttcccgac tggaaagcgg gcagtgagcg caacgcaatt
     7981 aatgtgagtt agctcactca ttaggcaccc caggctttac actttatgct tccggctcgt
     8041 atgttgtgtg gaattgtgag cggataacaa tttcacacag gaaacagcta tgaccatgat
     8101 tacgccaagc tctagctaga ggtcgaccaa ttctcatgtt tgacagctta tcatcgcaga
     8161 tccgggcaac gttgttgcca ttgctgcagg cgcagaactg gtaggtatgg aagatctata
     8221 cattgaatca atattggcaa ttagccatat tagtcattgg ttatatagca taaatcaata
     8281 ttggctattg gccattgcat acgttgtatc tatatcataa tatgtacatt tatattggct
     8341 catgtccaat atgaccgcca tgttgacatt gattattgac tagttattaa tagtaatcaa
     8401 ttacggggtc attagttcat agcccatata tggagttccg cgttacataa cttacggtaa
     8461 atggcccgcc tggctgaccg cccaacgacc cccgcccatt gacgtcaata atgacgtatg
     8521 ttcccatagt aacgccaata gggactttcc attgacgtca atgggtggag tatttacggt
     8581 aaactgccca cttggcagta catcaagtgt atcatatgcc aagtccgccc cctattgacg
     8641 tcaatgacgg taaatggccc gcctggcatt atgcccagta catgacctta cgggactttc
     8701 ctacttggca gtacatctac gtattagtca tcgctattac catggtgatg cggttttggc
     8761 agtacaccaa tgggcgtgga tagcggtttg actcacgggg atttccaagt ctccacccca
     8821 ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg ggactttcca aaatgtcgta
     8881 ataaccccgc cccgttgacg caaatgggcg gtaggcgtgt acggtgggag gtctatataa
     8941 gcagagctcg tttagtgaac cgtcagatcc tcactctctt ccgcatcgct gtctgcgagg
     9001 gccagctgtt gggctcgcgg ttgaggacaa actcttcgcg gtctttccag tactcttgga
     9061 tcggaaaccc gtcggcctcc gaacggtact ccgccaccga gggacctgag cgagtccgca
     9121 tcgaccggat cggaaaacct
//
