LOCUS       sequence-224591-        8480 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 8480)
  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 8480)
  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_1404900-COMP-blac-flag-his
FEATURES             Location/Qualifiers
     source          1..8480
                     /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             2014..2799
                     /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             2806..2871
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             2878..2895
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    2931..2986
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(2985..3004)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(5222..5240)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5308..5412
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5413..5481,5482..6273)
                     /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      6444..7032
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6933..6952
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7186..7203
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    7320..7341
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7356..7361,7362..7379,7380..7386)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7394..7410
                     /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     7399..7421
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7418..7434
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7418..7434
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     enhancer        7705..8084
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        8085..8288
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     8239..8259
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     8285..8309
                     /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 ggccgccaga
      421 accgagagca agaacatcct gaacaccgtg aagaacgacg tcaagaacgg caactaccag
      481 aacatgatga acttccccga catggacaag atcaaagagc tgatcaacca gaacaaggtg
      541 gaagaggccc gcgagctgtt caacgagatg cagaaacagt tcatgaagga aaagatggaa
      601 gccaacaaca acatgaacaa caagaagccc gccaagaaag ccatggacaa cagcaacctg
      661 ttcagcaacc tcatcaaccc caacgacctg caggaaatga tcaagttcta catgtacctg
      721 cagaatatcc tcaacagcaa gaacgaggcc agcgagaaga agatggtcaa gagcttcctg
      781 cgcaagacca tccagaagat gaaggacaac gagaacagcg aggaaaagaa gtccatcgac
      841 gacatcatca aggacaccga gggccacaat gagatgctgg aaaagctggc cggcctgatg
      901 aagatcaaca aagagaagct gaaagacgag caggtcaaaa acaagctgaa ccagatcctg
      961 atcggcatgc tcaagttcat cgactacacc aacaacgccg acatcaccaa cagcctgatg
     1021 gacgagatcc agatcgaaca ggtgcaggcc gaggacggca ccatcaagcc caaggtgcag
     1081 gtcaacattg ccaactccaa ggcccacctg gacatgctgc agaaagccac caacttcatg
     1141 ggcatcaaca tcgaccagga agaactgaag aagctgaccg tcaacaacaa gtggtacgag
     1201 tccttcctga acaacctgct gaactccgcc gacgaactcg gcgcgccgtc gacctccatc
     1261 acggcctata agagtgaggg ggagtcagcg gagttctcct tcccactcaa ccttggagag
     1321 gaaagcctgc agggagagtt gagatggaag gcagagaagg ctccttcttc ccagtcctgg
     1381 atcaccttct ccctaaagaa ccaaaaggtg tctgtgcaga agtctactag caaccccaag
     1441 ttccagctgt ccgaaacgct cccactcacc cttcagatac cccaggtctc ccttcagttt
     1501 gctggttctg gcaacctgac cctgactctg gacagaggga tactgtatca ggaagtgaac
     1561 ctggtggtga tgaaagtgac tcagcccgac agcaacactt tgacctgtga ggtgatggga
     1621 cccacctcac ccaagatgag actgatcttg aagcaggaga atcaggaggc cagggtctcc
     1681 aggcaggaga aagtgattca agtgcaggcc cctgaagcag gggtgtggca atgtctactg
     1741 agtgaaggtg aagaggtcaa gatggactcc aagatccagg ttttatccaa agggttgaat
     1801 tccggcggcg gctccggagg cggtaccggt ggcggcgacc tagccccaca gatgcttcga
     1861 gaactccagg agactaatgc ggcgctgcaa gacgtgagag agctcttgcg acagcaggtc
     1921 aaggagatca ccttcctgaa gaatacggtg atggaatgtg acgcctgcgg tctagacaga
     1981 aacctgcctc cactagcacc tctgggccca cacccagaaa cgctggtgaa agtaaaagat
     2041 gctgaagatc agttgggtgc acgagtgggt tacatcgaac tggatctcaa cagcggtaag
     2101 atccttgaga gttttcgccc cgaagaacgt tttccaatga tgagcacttt taaagttctg
     2161 ctatgtggcg cggtattatc ccgtattgac gccgggcaag agcaactcgg tcgccgcata
     2221 cactattctc agaatgactt ggttgagtac tcaccagtca cagaaaagca tcttacggat
     2281 ggcatgacag taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc
     2341 aacttacttc tgacaacgat cggaggaccg aaggagctaa ccgctttttt gcacaacatg
     2401 ggggatcatg taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac
     2461 gacgagcgtg acaccacgat gcctgtagca atggcaacaa cgttgcgcaa actattaact
     2521 ggcgaactac ttactctagc ttcccggcaa caattaatag actggatgga ggcggataaa
     2581 gttgcaggac cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct
     2641 ggagccggtg agcgtgggtc tcgcggtatc attgcagcac tggggccaga tggtaagccc
     2701 tcccgtatcg tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga
     2761 cagatcgctg agataggtgc ctcactgatt aagcattggg gccctgacta caaagaccat
     2821 gacggtgatt ataaagatca tgacatcgat tacaaggatg acgatgacaa gggccctcac
     2881 caccaccatc accactaata ataaggatcc cccgacctcg acctctggct aataaaggaa
     2941 atttattttc attgcaatag tgtgttggaa ttttttgtgt ctctcactcg gaaggacata
     3001 tgggagggca aatcatttgg tcgagatccc tcggagatct ctagctagag gatcgatccc
     3061 cgccccggac gaactaaacc tgactacgac atctctgccc cttcttcgcg gggcagtgca
     3121 tgtaatccct tcagttggtt ggtacaactt gccaactggg ccctgttcca catgtgacac
     3181 ggggggggac caaacacaaa ggggttctct gactgtagtt gacatcctta taaatggatg
     3241 tgcacatttg ccaacactga gtggctttca tcctggagca gactttgcag tctgtggact
     3301 gcaacacaac attgccttta tgtgtaactc ttggctgaag ctcttacacc aatgctgggg
     3361 gacatgtacc tcccaggggc ccaggaagac tacgggaggc tacaccaacg tcaatcagag
     3421 gggcctgtgt agctaccgat aagcggaccc tcaagagggc attagcaata gtgtttataa
     3481 ggcccccttg ttaaccctaa acgggtagca tatgcttccc gggtagtagt atatactatc
     3541 cagactaacc ctaattcaat agcatatgtt acccaacggg aagcatatgc tatcgaatta
     3601 gggttagtaa aagggtccta aggaacagcg atatctccca ccccatgagc tgtcacggtt
     3661 ttatttacat ggggtcagga ttccacgagg gtagtgaacc attttagtca caagggcagt
     3721 ggctgaagat caaggagcgg gcagtgaact ctcctgaatc ttcgcctgct tcttcattct
     3781 ccttcgttta gctaatagaa taactgctga gttgtgaaca gtaaggtgta tgtgaggtgc
     3841 tcgaaaacaa ggtttcaggt gacgccccca gaataaaatt tggacggggg gttcagtggt
     3901 ggcattgtgc tatgacacca atataaccct cacaaacccc ttgggcaata aatactagtg
     3961 taggaatgaa acattctgaa tatctttaac aatagaaatc catggggtgg ggacaagccg
     4021 taaagactgg atgtccatct cacacgaatt tatggctatg ggcaacacat aatcctagtg
     4081 caatatgata ctggggttat taagatgtgt cccaggcagg gaccaagaca ggtgaaccat
     4141 gttgttacac tctatttgta acaaggggaa agagagtgga cgccgacagc agcggactcc
     4201 actggttgtc tctaacaccc ccgaaaatta aacggggctc cacgccaatg gggcccataa
     4261 acaaagacaa gtggccactc ttttttttga aattgtggag tgggggcacg cgtcagcccc
     4321 cacacgccgc cctgcggttt tggactgtaa aataagggtg taataacttg gctgattgta
     4381 accccgctaa ccactgcggt caaaccactt gcccacaaaa ccactaatgg caccccgggg
     4441 aatacctgca taagtaggtg ggcgggccaa gataggggcg cgattgctgc gatctggagg
     4501 acaaattaca cacacttgcg cctgagcgcc aagcacaggg ttgttggtcc tcatattcac
     4561 gaggtcgctg agagcacggt gggctaatgt tgccatgggt agcatatact acccaaatat
     4621 ctggatagca tatgctatcc taatctatat ctgggtagca taggctatcc taatctatat
     4681 ctgggtagca tatgctatcc taatctatat ctgggtagta tatgctatcc taatttatat
     4741 ctgggtagca taggctatcc taatctatat ctgggtagca tatgctatcc taatctatat
     4801 ctgggtagta tatgctatcc taatctgtat ccgggtagca tatgctatcc taatagagat
     4861 tagggtagta tatgctatcc taatttatat ctgggtagca tatactaccc aaatatctgg
     4921 atagcatatg ctatcctaat ctatatctgg gtagcatatg ctatcctaat ctatatctgg
     4981 gtagcatagg ctatcctaat ctatatctgg gtagcatatg ctatcctaat ctatatctgg
     5041 gtagtatatg ctatcctaat ttatatctgg gtagcatagg ctatcctaat ctatatctgg
     5101 gtagcatatg ctatcctaat ctatatctgg gtagtatatg ctatcctaat ctgtatccgg
     5161 gtagcatatg ctatcctcat gataagctgt caaacatgag aattttcttg aagacgaaag
     5221 ggcctcgtga tacgcctatt tttataggtt aatgtcatga taataatggt ttcttagacg
     5281 tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata
     5341 cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga
     5401 aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc ttattccctt ttttgcggca
     5461 ttttgccttc ctgtttttgc tcacccagaa acgctggtga aagtaaaaga tgctgaagat
     5521 cagttgggtg cacgagtggg ttacatcgaa ctggatctca acagcggtaa gatccttgag
     5581 agttttcgcc ccgaagaacg ttttccaatg atgagcactt ttaaagttct gctatgtggc
     5641 gcggtattat cccgtgttga cgccgggcaa gagcaactcg gtcgccgcat acactattct
     5701 cagaatgact tggttgagta ctcaccagtc acagaaaagc atcttacgga tggcatgaca
     5761 gtaagagaat tatgcagtgc tgccataacc atgagtgata acactgcggc caacttactt
     5821 ctgacaacga tcggaggacc gaaggagcta accgcttttt tgcacaacat gggggatcat
     5881 gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt
     5941 gacaccacga tgcctgcagc aatggcaaca acgttgcgca aactattaac tggcgaacta
     6001 cttactctag cttcccggca acaattaata gactggatgg aggcggataa agttgcagga
     6061 ccacttctgc gctcggccct tccggctggc tggtttattg ctgataaatc tggagccggt
     6121 gagcgtgggt ctcgcggtat cattgcagca ctggggccag atggtaagcc ctcccgtatc
     6181 gtagttatct acacgacggg gagtcaggca actatggatg aacgaaatag acagatcgct
     6241 gagataggtg cctcactgat taagcattgg taactgtcag accaagttta ctcatatata
     6301 ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt
     6361 gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc
     6421 gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg
     6481 caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact
     6541 ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt tcttctagtg
     6601 tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg
     6661 ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac
     6721 tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca
     6781 cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga
     6841 gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc
     6901 ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct
     6961 gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg
     7021 agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct
     7081 tttgctcaca tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc
     7141 tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc
     7201 gaggaagcgg aagagcgccc aatacgcaaa ccgcctctcc ccgcgcgttg gccgattcat
     7261 taatgcagct ggcacgacag gtttcccgac tggaaagcgg gcagtgagcg caacgcaatt
     7321 aatgtgagtt agctcactca ttaggcaccc caggctttac actttatgct tccggctcgt
     7381 atgttgtgtg gaattgtgag cggataacaa tttcacacag gaaacagcta tgaccatgat
     7441 tacgccaagc tctagctaga ggtcgaccaa ttctcatgtt tgacagctta tcatcgcaga
     7501 tccgggcaac gttgttgcca ttgctgcagg cgcagaactg gtaggtatgg aagatctata
     7561 cattgaatca atattggcaa ttagccatat tagtcattgg ttatatagca taaatcaata
     7621 ttggctattg gccattgcat acgttgtatc tatatcataa tatgtacatt tatattggct
     7681 catgtccaat atgaccgcca tgttgacatt gattattgac tagttattaa tagtaatcaa
     7741 ttacggggtc attagttcat agcccatata tggagttccg cgttacataa cttacggtaa
     7801 atggcccgcc tggctgaccg cccaacgacc cccgcccatt gacgtcaata atgacgtatg
     7861 ttcccatagt aacgccaata gggactttcc attgacgtca atgggtggag tatttacggt
     7921 aaactgccca cttggcagta catcaagtgt atcatatgcc aagtccgccc cctattgacg
     7981 tcaatgacgg taaatggccc gcctggcatt atgcccagta catgacctta cgggactttc
     8041 ctacttggca gtacatctac gtattagtca tcgctattac catggtgatg cggttttggc
     8101 agtacaccaa tgggcgtgga tagcggtttg actcacgggg atttccaagt ctccacccca
     8161 ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg ggactttcca aaatgtcgta
     8221 ataaccccgc cccgttgacg caaatgggcg gtaggcgtgt acggtgggag gtctatataa
     8281 gcagagctcg tttagtgaac cgtcagatcc tcactctctt ccgcatcgct gtctgcgagg
     8341 gccagctgtt gggctcgcgg ttgaggacaa actcttcgcg gtctttccag tactcttgga
     8401 tcggaaaccc gtcggcctcc gaacggtact ccgccaccga gggacctgag cgagtccgca
     8461 tcgaccggat cggaaaacct
//
