LOCUS       sequence-208810-        8320 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 8320)
  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 8320)
  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_0721100-COMP-blac-flag-his
FEATURES             Location/Qualifiers
     source          1..8320
                     /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             1852..2637
                     /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             2644..2709
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             2716..2733
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    2771..2826
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(2825..2844)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(5062..5080)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5148..5252
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5253..5321,5322..6113)
                     /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      6284..6872
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6773..6792
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7026..7043
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    7160..7181
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7196..7201,7202..7219,7220..7226)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7234..7250
                     /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     7239..7261
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7258..7274
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7258..7274
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     enhancer        7545..7924
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        7925..8128
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     8079..8099
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     8125..8149
                     /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 ggcaaccaga tctctaagtg cgtgttctac gacaagcgga acaaggacct gaaacaggtg
      481 ttctatgtgc ccaccaacga cgtgaacatg tacaaagaga caatccggaa gctgctgagc
      541 agagagatca gcctgctgga agaacccctg aacgagaaca gcgacagcca caacatggcc
      601 aacaagaaaa acgtggccaa cttcgaggaa atcaagaaga acgacgagaa caacaaccgc
      661 aagcgcaaag agatggaaga gaagctgaag tacatcgaga agtaccggaa ctacgccaag
      721 ggcgtgctga agaacgtgga attcaagtgc gagaatgtgg acgacctgaa gtccaaagtg
      781 aagtgcaacc tgtgcgtgta caccgagatc gtgaacaaca agcacatccg gcagggccag
      841 atcatccaga tcttcatgaa cgagtgtatg tacggcttct actactacga caacatcaac
      901 cagtacaccc agaacaatga gtacaacaac aaggccctca aggactacaa cagcgcccag
      961 aagctggacg acatctactt caatatcgcc aaggacatca gcgagggcaa cgtgaacggc
     1021 atcaatttct tcaagcagca gagcagcaag taccaccccg tgtgctacgt gaccgaaggc
     1081 gcgccgtcga cctccatcac ggcctataag agtgaggggg agtcagcgga gttctccttc
     1141 ccactcaacc ttggagagga aagcctgcag ggagagttga gatggaaggc agagaaggct
     1201 ccttcttccc agtcctggat caccttctcc ctaaagaacc aaaaggtgtc tgtgcagaag
     1261 tctactagca accccaagtt ccagctgtcc gaaacgctcc cactcaccct tcagataccc
     1321 caggtctccc ttcagtttgc tggttctggc aacctgaccc tgactctgga cagagggata
     1381 ctgtatcagg aagtgaacct ggtggtgatg aaagtgactc agcccgacag caacactttg
     1441 acctgtgagg tgatgggacc cacctcaccc aagatgagac tgatcttgaa gcaggagaat
     1501 caggaggcca gggtctccag gcaggagaaa gtgattcaag tgcaggcccc tgaagcaggg
     1561 gtgtggcaat gtctactgag tgaaggtgaa gaggtcaaga tggactccaa gatccaggtt
     1621 ttatccaaag ggttgaattc cggcggcggc tccggaggcg gtaccggtgg cggcgaccta
     1681 gccccacaga tgcttcgaga actccaggag actaatgcgg cgctgcaaga cgtgagagag
     1741 ctcttgcgac agcaggtcaa ggagatcacc ttcctgaaga atacggtgat ggaatgtgac
     1801 gcctgcggtc tagacagaaa cctgcctcca ctagcacctc tgggcccaca cccagaaacg
     1861 ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac gagtgggtta catcgaactg
     1921 gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgttt tccaatgatg
     1981 agcactttta aagttctgct atgtggcgcg gtattatccc gtattgacgc cgggcaagag
     2041 caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtactc accagtcaca
     2101 gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc cataaccatg
     2161 agtgataaca ctgcggccaa cttacttctg acaacgatcg gaggaccgaa ggagctaacc
     2221 gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga accggagctg
     2281 aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg
     2341 ttgcgcaaac tattaactgg cgaactactt actctagctt cccggcaaca attaatagac
     2401 tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc ggctggctgg
     2461 tttattgctg ataaatctgg agccggtgag cgtgggtctc gcggtatcat tgcagcactg
     2521 gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag tcaggcaact
     2581 atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggggc
     2641 cctgactaca aagaccatga cggtgattat aaagatcatg acatcgatta caaggatgac
     2701 gatgacaagg gccctcacca ccaccatcac cactaattaa ttaaggatcc cccgacctcg
     2761 acctctggct aataaaggaa atttattttc attgcaatag tgtgttggaa ttttttgtgt
     2821 ctctcactcg gaaggacata tgggagggca aatcatttgg tcgagatccc tcggagatct
     2881 ctagctagag gatcgatccc cgccccggac gaactaaacc tgactacgac atctctgccc
     2941 cttcttcgcg gggcagtgca tgtaatccct tcagttggtt ggtacaactt gccaactggg
     3001 ccctgttcca catgtgacac ggggggggac caaacacaaa ggggttctct gactgtagtt
     3061 gacatcctta taaatggatg tgcacatttg ccaacactga gtggctttca tcctggagca
     3121 gactttgcag tctgtggact gcaacacaac attgccttta tgtgtaactc ttggctgaag
     3181 ctcttacacc aatgctgggg gacatgtacc tcccaggggc ccaggaagac tacgggaggc
     3241 tacaccaacg tcaatcagag gggcctgtgt agctaccgat aagcggaccc tcaagagggc
     3301 attagcaata gtgtttataa ggcccccttg ttaaccctaa acgggtagca tatgcttccc
     3361 gggtagtagt atatactatc cagactaacc ctaattcaat agcatatgtt acccaacggg
     3421 aagcatatgc tatcgaatta gggttagtaa aagggtccta aggaacagcg atatctccca
     3481 ccccatgagc tgtcacggtt ttatttacat ggggtcagga ttccacgagg gtagtgaacc
     3541 attttagtca caagggcagt ggctgaagat caaggagcgg gcagtgaact ctcctgaatc
     3601 ttcgcctgct tcttcattct ccttcgttta gctaatagaa taactgctga gttgtgaaca
     3661 gtaaggtgta tgtgaggtgc tcgaaaacaa ggtttcaggt gacgccccca gaataaaatt
     3721 tggacggggg gttcagtggt ggcattgtgc tatgacacca atataaccct cacaaacccc
     3781 ttgggcaata aatactagtg taggaatgaa acattctgaa tatctttaac aatagaaatc
     3841 catggggtgg ggacaagccg taaagactgg atgtccatct cacacgaatt tatggctatg
     3901 ggcaacacat aatcctagtg caatatgata ctggggttat taagatgtgt cccaggcagg
     3961 gaccaagaca ggtgaaccat gttgttacac tctatttgta acaaggggaa agagagtgga
     4021 cgccgacagc agcggactcc actggttgtc tctaacaccc ccgaaaatta aacggggctc
     4081 cacgccaatg gggcccataa acaaagacaa gtggccactc ttttttttga aattgtggag
     4141 tgggggcacg cgtcagcccc cacacgccgc cctgcggttt tggactgtaa aataagggtg
     4201 taataacttg gctgattgta accccgctaa ccactgcggt caaaccactt gcccacaaaa
     4261 ccactaatgg caccccgggg aatacctgca taagtaggtg ggcgggccaa gataggggcg
     4321 cgattgctgc gatctggagg acaaattaca cacacttgcg cctgagcgcc aagcacaggg
     4381 ttgttggtcc tcatattcac gaggtcgctg agagcacggt gggctaatgt tgccatgggt
     4441 agcatatact acccaaatat ctggatagca tatgctatcc taatctatat ctgggtagca
     4501 taggctatcc taatctatat ctgggtagca tatgctatcc taatctatat ctgggtagta
     4561 tatgctatcc taatttatat ctgggtagca taggctatcc taatctatat ctgggtagca
     4621 tatgctatcc taatctatat ctgggtagta tatgctatcc taatctgtat ccgggtagca
     4681 tatgctatcc taatagagat tagggtagta tatgctatcc taatttatat ctgggtagca
     4741 tatactaccc aaatatctgg atagcatatg ctatcctaat ctatatctgg gtagcatatg
     4801 ctatcctaat ctatatctgg gtagcatagg ctatcctaat ctatatctgg gtagcatatg
     4861 ctatcctaat ctatatctgg gtagtatatg ctatcctaat ttatatctgg gtagcatagg
     4921 ctatcctaat ctatatctgg gtagcatatg ctatcctaat ctatatctgg gtagtatatg
     4981 ctatcctaat ctgtatccgg gtagcatatg ctatcctcat gataagctgt caaacatgag
     5041 aattttcttg aagacgaaag ggcctcgtga tacgcctatt tttataggtt aatgtcatga
     5101 taataatggt ttcttagacg tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta
     5161 tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat
     5221 aaatgcttca ataatattga aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc
     5281 ttattccctt ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctggtga
     5341 aagtaaaaga tgctgaagat cagttgggtg cacgagtggg ttacatcgaa ctggatctca
     5401 acagcggtaa gatccttgag agttttcgcc ccgaagaacg ttttccaatg atgagcactt
     5461 ttaaagttct gctatgtggc gcggtattat cccgtgttga cgccgggcaa gagcaactcg
     5521 gtcgccgcat acactattct cagaatgact tggttgagta ctcaccagtc acagaaaagc
     5581 atcttacgga tggcatgaca gtaagagaat tatgcagtgc tgccataacc atgagtgata
     5641 acactgcggc caacttactt ctgacaacga tcggaggacc gaaggagcta accgcttttt
     5701 tgcacaacat gggggatcat gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag
     5761 ccataccaaa cgacgagcgt gacaccacga tgcctgcagc aatggcaaca acgttgcgca
     5821 aactattaac tggcgaacta cttactctag cttcccggca acaattaata gactggatgg
     5881 aggcggataa agttgcagga ccacttctgc gctcggccct tccggctggc tggtttattg
     5941 ctgataaatc tggagccggt gagcgtgggt ctcgcggtat cattgcagca ctggggccag
     6001 atggtaagcc ctcccgtatc gtagttatct acacgacggg gagtcaggca actatggatg
     6061 aacgaaatag acagatcgct gagataggtg cctcactgat taagcattgg taactgtcag
     6121 accaagttta ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga
     6181 tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt
     6241 tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc
     6301 tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc
     6361 cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac
     6421 caaatactgt tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac
     6481 cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt
     6541 cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct
     6601 gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat
     6661 acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt
     6721 atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg
     6781 cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt
     6841 gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt
     6901 tcctggcctt ttgctggcct tttgctcaca tgttctttcc tgcgttatcc cctgattctg
     6961 tggataaccg tattaccgcc tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg
     7021 agcgcagcga gtcagtgagc gaggaagcgg aagagcgccc aatacgcaaa ccgcctctcc
     7081 ccgcgcgttg gccgattcat taatgcagct ggcacgacag gtttcccgac tggaaagcgg
     7141 gcagtgagcg caacgcaatt aatgtgagtt agctcactca ttaggcaccc caggctttac
     7201 actttatgct tccggctcgt atgttgtgtg gaattgtgag cggataacaa tttcacacag
     7261 gaaacagcta tgaccatgat tacgccaagc tctagctaga ggtcgaccaa ttctcatgtt
     7321 tgacagctta tcatcgcaga tccgggcaac gttgttgcca ttgctgcagg cgcagaactg
     7381 gtaggtatgg aagatctata cattgaatca atattggcaa ttagccatat tagtcattgg
     7441 ttatatagca taaatcaata ttggctattg gccattgcat acgttgtatc tatatcataa
     7501 tatgtacatt tatattggct catgtccaat atgaccgcca tgttgacatt gattattgac
     7561 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg
     7621 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
     7681 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
     7741 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
     7801 aagtccgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
     7861 catgacctta cgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
     7921 catggtgatg cggttttggc agtacaccaa tgggcgtgga tagcggtttg actcacgggg
     7981 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
     8041 ggactttcca aaatgtcgta ataaccccgc cccgttgacg caaatgggcg gtaggcgtgt
     8101 acggtgggag gtctatataa gcagagctcg tttagtgaac cgtcagatcc tcactctctt
     8161 ccgcatcgct gtctgcgagg gccagctgtt gggctcgcgg ttgaggacaa actcttcgcg
     8221 gtctttccag tactcttgga tcggaaaccc gtcggcctcc gaacggtact ccgccaccga
     8281 gggacctgag cgagtccgca tcgaccggat cggaaaacct
//
