LOCUS       sequence-208817-        8410 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 8410)
  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 8410)
  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_1012200-COMP-blac-flag-his
FEATURES             Location/Qualifiers
     source          1..8410
                     /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             1942..2727
                     /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             2734..2799
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             2806..2823
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    2861..2916
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(2915..2934)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(5152..5170)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5238..5342
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5343..5411,5412..6203)
                     /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      6374..6962
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6863..6882
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7116..7133
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    7250..7271
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7286..7291,7292..7309,7310..7316)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7324..7340
                     /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     7329..7351
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7348..7364
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7348..7364
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     enhancer        7635..8014
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        8015..8218
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     8169..8189
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     8215..8239
                     /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 ggccgccgag
      421 aacgtggaca ccttcaagtg caactacgcc aagaagaaga aaaacggcca ccacatcaag
      481 cggcacatca ccaacgacga ggaaaagaag aaagaatact ccttcctgat gctgggcaaa
      541 gagaacgagg aagaaaacaa ggaaaacaaa gagaataacc agaacgtcaa caaagacaac
      601 aacgataaca acaacaatga gaaaaagaat gaggaacaga accataacaa cgagaagaag
      661 caggaagaag tcatcaacaa taacaataat aacgtcgaga acaagaaaga agaggaaaac
      721 cacaataacg acaagaaaac cgacgagcag aaccacgtga acgaaaaaaa acaggaaggc
      781 gagaacaaca agcacaagaa ggacgacacc atccctccag agaagaagat taacaaagaa
      841 aacctgctcg agtacggcac ccacgacaaa gagggccact tcatccccag ctacaagacc
      901 ctgaccgacg agatcctgag caccaacaac gccatggaaa aggccagcag cttcctgaag
      961 atcgcctgca gccacgtgat gaagctgatc gagttcatcc ccgagagcaa gctgagcagc
     1021 cagtacatca aggtggacaa caagaacatc tacctgaagg atatcgccgt cgagtgccag
     1081 aatatctact tcaacctcga gaagttcagc atgaccctga tcgtgttcaa cagcaagatc
     1141 aacaagttca tctacagcca ggaaaaaggc gcgccgtcga cctccatcac ggcctataag
     1201 agtgaggggg agtcagcgga gttctccttc ccactcaacc ttggagagga aagcctgcag
     1261 ggagagttga gatggaaggc agagaaggct ccttcttccc agtcctggat caccttctcc
     1321 ctaaagaacc aaaaggtgtc tgtgcagaag tctactagca accccaagtt ccagctgtcc
     1381 gaaacgctcc cactcaccct tcagataccc caggtctccc ttcagtttgc tggttctggc
     1441 aacctgaccc tgactctgga cagagggata ctgtatcagg aagtgaacct ggtggtgatg
     1501 aaagtgactc agcccgacag caacactttg acctgtgagg tgatgggacc cacctcaccc
     1561 aagatgagac tgatcttgaa gcaggagaat caggaggcca gggtctccag gcaggagaaa
     1621 gtgattcaag tgcaggcccc tgaagcaggg gtgtggcaat gtctactgag tgaaggtgaa
     1681 gaggtcaaga tggactccaa gatccaggtt ttatccaaag ggttgaattc cggcggcggc
     1741 tccggaggcg gtaccggtgg cggcgaccta gccccacaga tgcttcgaga actccaggag
     1801 actaatgcgg cgctgcaaga cgtgagagag ctcttgcgac agcaggtcaa ggagatcacc
     1861 ttcctgaaga atacggtgat ggaatgtgac gcctgcggtc tagacagaaa cctgcctcca
     1921 ctagcacctc tgggcccaca cccagaaacg ctggtgaaag taaaagatgc tgaagatcag
     1981 ttgggtgcac gagtgggtta catcgaactg gatctcaaca gcggtaagat ccttgagagt
     2041 tttcgccccg aagaacgttt tccaatgatg agcactttta aagttctgct atgtggcgcg
     2101 gtattatccc gtattgacgc cgggcaagag caactcggtc gccgcataca ctattctcag
     2161 aatgacttgg ttgagtactc accagtcaca gaaaagcatc ttacggatgg catgacagta
     2221 agagaattat gcagtgctgc cataaccatg agtgataaca ctgcggccaa cttacttctg
     2281 acaacgatcg gaggaccgaa ggagctaacc gcttttttgc acaacatggg ggatcatgta
     2341 actcgccttg atcgttggga accggagctg aatgaagcca taccaaacga cgagcgtgac
     2401 accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg cgaactactt
     2461 actctagctt cccggcaaca attaatagac tggatggagg cggataaagt tgcaggacca
     2521 cttctgcgct cggcccttcc ggctggctgg tttattgctg ataaatctgg agccggtgag
     2581 cgtgggtctc gcggtatcat tgcagcactg gggccagatg gtaagccctc ccgtatcgta
     2641 gttatctaca cgacggggag tcaggcaact atggatgaac gaaatagaca gatcgctgag
     2701 ataggtgcct cactgattaa gcattggggc cctgactaca aagaccatga cggtgattat
     2761 aaagatcatg acatcgatta caaggatgac gatgacaagg gccctcacca ccaccatcac
     2821 cactaattaa ttaaggatcc cccgacctcg acctctggct aataaaggaa atttattttc
     2881 attgcaatag tgtgttggaa ttttttgtgt ctctcactcg gaaggacata tgggagggca
     2941 aatcatttgg tcgagatccc tcggagatct ctagctagag gatcgatccc cgccccggac
     3001 gaactaaacc tgactacgac atctctgccc cttcttcgcg gggcagtgca tgtaatccct
     3061 tcagttggtt ggtacaactt gccaactggg ccctgttcca catgtgacac ggggggggac
     3121 caaacacaaa ggggttctct gactgtagtt gacatcctta taaatggatg tgcacatttg
     3181 ccaacactga gtggctttca tcctggagca gactttgcag tctgtggact gcaacacaac
     3241 attgccttta tgtgtaactc ttggctgaag ctcttacacc aatgctgggg gacatgtacc
     3301 tcccaggggc ccaggaagac tacgggaggc tacaccaacg tcaatcagag gggcctgtgt
     3361 agctaccgat aagcggaccc tcaagagggc attagcaata gtgtttataa ggcccccttg
     3421 ttaaccctaa acgggtagca tatgcttccc gggtagtagt atatactatc cagactaacc
     3481 ctaattcaat agcatatgtt acccaacggg aagcatatgc tatcgaatta gggttagtaa
     3541 aagggtccta aggaacagcg atatctccca ccccatgagc tgtcacggtt ttatttacat
     3601 ggggtcagga ttccacgagg gtagtgaacc attttagtca caagggcagt ggctgaagat
     3661 caaggagcgg gcagtgaact ctcctgaatc ttcgcctgct tcttcattct ccttcgttta
     3721 gctaatagaa taactgctga gttgtgaaca gtaaggtgta tgtgaggtgc tcgaaaacaa
     3781 ggtttcaggt gacgccccca gaataaaatt tggacggggg gttcagtggt ggcattgtgc
     3841 tatgacacca atataaccct cacaaacccc ttgggcaata aatactagtg taggaatgaa
     3901 acattctgaa tatctttaac aatagaaatc catggggtgg ggacaagccg taaagactgg
     3961 atgtccatct cacacgaatt tatggctatg ggcaacacat aatcctagtg caatatgata
     4021 ctggggttat taagatgtgt cccaggcagg gaccaagaca ggtgaaccat gttgttacac
     4081 tctatttgta acaaggggaa agagagtgga cgccgacagc agcggactcc actggttgtc
     4141 tctaacaccc ccgaaaatta aacggggctc cacgccaatg gggcccataa acaaagacaa
     4201 gtggccactc ttttttttga aattgtggag tgggggcacg cgtcagcccc cacacgccgc
     4261 cctgcggttt tggactgtaa aataagggtg taataacttg gctgattgta accccgctaa
     4321 ccactgcggt caaaccactt gcccacaaaa ccactaatgg caccccgggg aatacctgca
     4381 taagtaggtg ggcgggccaa gataggggcg cgattgctgc gatctggagg acaaattaca
     4441 cacacttgcg cctgagcgcc aagcacaggg ttgttggtcc tcatattcac gaggtcgctg
     4501 agagcacggt gggctaatgt tgccatgggt agcatatact acccaaatat ctggatagca
     4561 tatgctatcc taatctatat ctgggtagca taggctatcc taatctatat ctgggtagca
     4621 tatgctatcc taatctatat ctgggtagta tatgctatcc taatttatat ctgggtagca
     4681 taggctatcc taatctatat ctgggtagca tatgctatcc taatctatat ctgggtagta
     4741 tatgctatcc taatctgtat ccgggtagca tatgctatcc taatagagat tagggtagta
     4801 tatgctatcc taatttatat ctgggtagca tatactaccc aaatatctgg atagcatatg
     4861 ctatcctaat ctatatctgg gtagcatatg ctatcctaat ctatatctgg gtagcatagg
     4921 ctatcctaat ctatatctgg gtagcatatg ctatcctaat ctatatctgg gtagtatatg
     4981 ctatcctaat ttatatctgg gtagcatagg ctatcctaat ctatatctgg gtagcatatg
     5041 ctatcctaat ctatatctgg gtagtatatg ctatcctaat ctgtatccgg gtagcatatg
     5101 ctatcctcat gataagctgt caaacatgag aattttcttg aagacgaaag ggcctcgtga
     5161 tacgcctatt tttataggtt aatgtcatga taataatggt ttcttagacg tcaggtggca
     5221 cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata
     5281 tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga
     5341 gtatgagtat tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc
     5401 ctgtttttgc tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg
     5461 cacgagtggg ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc
     5521 ccgaagaacg ttttccaatg atgagcactt ttaaagttct gctatgtggc gcggtattat
     5581 cccgtgttga cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact
     5641 tggttgagta ctcaccagtc acagaaaagc atcttacgga tggcatgaca gtaagagaat
     5701 tatgcagtgc tgccataacc atgagtgata acactgcggc caacttactt ctgacaacga
     5761 tcggaggacc gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc
     5821 ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt gacaccacga
     5881 tgcctgcagc aatggcaaca acgttgcgca aactattaac tggcgaacta cttactctag
     5941 cttcccggca acaattaata gactggatgg aggcggataa agttgcagga ccacttctgc
     6001 gctcggccct tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtgggt
     6061 ctcgcggtat cattgcagca ctggggccag atggtaagcc ctcccgtatc gtagttatct
     6121 acacgacggg gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg
     6181 cctcactgat taagcattgg taactgtcag accaagttta ctcatatata ctttagattg
     6241 atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca
     6301 tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga
     6361 tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa
     6421 aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga
     6481 aggtaactgg cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt
     6541 taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt
     6601 taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat
     6661 agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct
     6721 tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca
     6781 cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag
     6841 agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc
     6901 gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga
     6961 aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca
     7021 tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc tttgagtgag
     7081 ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg
     7141 aagagcgccc aatacgcaaa ccgcctctcc ccgcgcgttg gccgattcat taatgcagct
     7201 ggcacgacag gtttcccgac tggaaagcgg gcagtgagcg caacgcaatt aatgtgagtt
     7261 agctcactca ttaggcaccc caggctttac actttatgct tccggctcgt atgttgtgtg
     7321 gaattgtgag cggataacaa tttcacacag gaaacagcta tgaccatgat tacgccaagc
     7381 tctagctaga ggtcgaccaa ttctcatgtt tgacagctta tcatcgcaga tccgggcaac
     7441 gttgttgcca ttgctgcagg cgcagaactg gtaggtatgg aagatctata cattgaatca
     7501 atattggcaa ttagccatat tagtcattgg ttatatagca taaatcaata ttggctattg
     7561 gccattgcat acgttgtatc tatatcataa tatgtacatt tatattggct catgtccaat
     7621 atgaccgcca tgttgacatt gattattgac tagttattaa tagtaatcaa ttacggggtc
     7681 attagttcat agcccatata tggagttccg cgttacataa cttacggtaa atggcccgcc
     7741 tggctgaccg cccaacgacc cccgcccatt gacgtcaata atgacgtatg ttcccatagt
     7801 aacgccaata gggactttcc attgacgtca atgggtggag tatttacggt aaactgccca
     7861 cttggcagta catcaagtgt atcatatgcc aagtccgccc cctattgacg tcaatgacgg
     7921 taaatggccc gcctggcatt atgcccagta catgacctta cgggactttc ctacttggca
     7981 gtacatctac gtattagtca tcgctattac catggtgatg cggttttggc agtacaccaa
     8041 tgggcgtgga tagcggtttg actcacgggg atttccaagt ctccacccca ttgacgtcaa
     8101 tgggagtttg ttttggcacc aaaatcaacg ggactttcca aaatgtcgta ataaccccgc
     8161 cccgttgacg caaatgggcg gtaggcgtgt acggtgggag gtctatataa gcagagctcg
     8221 tttagtgaac cgtcagatcc tcactctctt ccgcatcgct gtctgcgagg gccagctgtt
     8281 gggctcgcgg ttgaggacaa actcttcgcg gtctttccag tactcttgga tcggaaaccc
     8341 gtcggcctcc gaacggtact ccgccaccga gggacctgag cgagtccgca tcgaccggat
     8401 cggaaaacct
//
