LOCUS       sequence-224268-        8141 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 8141)
  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 8141)
  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_1333300-COMP-blac-flag-his
FEATURES             Location/Qualifiers
     source          1..8141
                     /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             1675..2460
                     /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             2467..2532
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             2539..2556
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    2592..2647
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(2646..2665)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(4883..4901)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4969..5073
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5074..5142,5143..5934)
                     /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      6105..6693
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6594..6613
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6847..6864
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    6981..7002
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7017..7022,7023..7040,7041..7047)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7055..7071
                     /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     7060..7082
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7079..7095
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7079..7095
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     enhancer        7366..7745
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        7746..7949
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     7900..7920
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     7946..7970
                     /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 ggccgcctac
      421 aacggcctgg aaatctacat caagatgaag ggcaagaaac tgaagtgcat caaagagcgg
      481 atcaacaagg acaccctggt catcagcaag ttcaagacca accacaagaa cgccctggtg
      541 tccctgtaca tctacgacgt ggacgtgaac gagaagaact tccacaccca gcccaagctg
      601 cccatcttcg agacagtgaa caagcacgac atcaagaccg ccttcaccac cttctacagc
      661 agcagctaca gcttctgtgc ctacaacaac gccgacaagc tgatcgaggt gtacttcgag
      721 attaagtacg gcaccgaggc ccgggactac gcccaggtgg ccaagaccga gcacctgaac
      781 cagggcaccc tgtacctgaa gcagatcctg gaccagatga acatcttcca cgtgaacctg
      841 aagaggatcc gcagcgacga ggacaaagag aagaaggccc acgataagct gaacgacgcc
      901 ggcgcgccgt cgacctccat cacggcctat aagagtgagg gggagtcagc ggagttctcc
      961 ttcccactca accttggaga ggaaagcctg cagggagagt tgagatggaa ggcagagaag
     1021 gctccttctt cccagtcctg gatcaccttc tccctaaaga accaaaaggt gtctgtgcag
     1081 aagtctacta gcaaccccaa gttccagctg tccgaaacgc tcccactcac ccttcagata
     1141 ccccaggtct cccttcagtt tgctggttct ggcaacctga ccctgactct ggacagaggg
     1201 atactgtatc aggaagtgaa cctggtggtg atgaaagtga ctcagcccga cagcaacact
     1261 ttgacctgtg aggtgatggg acccacctca cccaagatga gactgatctt gaagcaggag
     1321 aatcaggagg ccagggtctc caggcaggag aaagtgattc aagtgcaggc ccctgaagca
     1381 ggggtgtggc aatgtctact gagtgaaggt gaagaggtca agatggactc caagatccag
     1441 gttttatcca aagggttgaa ttccggcggc ggctccggag gcggtaccgg tggcggcgac
     1501 ctagccccac agatgcttcg agaactccag gagactaatg cggcgctgca agacgtgaga
     1561 gagctcttgc gacagcaggt caaggagatc accttcctga agaatacggt gatggaatgt
     1621 gacgcctgcg gtctagacag aaacctgcct ccactagcac ctctgggccc acacccagaa
     1681 acgctggtga aagtaaaaga tgctgaagat cagttgggtg cacgagtggg ttacatcgaa
     1741 ctggatctca acagcggtaa gatccttgag agttttcgcc ccgaagaacg ttttccaatg
     1801 atgagcactt ttaaagttct gctatgtggc gcggtattat cccgtattga cgccgggcaa
     1861 gagcaactcg gtcgccgcat acactattct cagaatgact tggttgagta ctcaccagtc
     1921 acagaaaagc atcttacgga tggcatgaca gtaagagaat tatgcagtgc tgccataacc
     1981 atgagtgata acactgcggc caacttactt ctgacaacga tcggaggacc gaaggagcta
     2041 accgcttttt tgcacaacat gggggatcat gtaactcgcc ttgatcgttg ggaaccggag
     2101 ctgaatgaag ccataccaaa cgacgagcgt gacaccacga tgcctgtagc aatggcaaca
     2161 acgttgcgca aactattaac tggcgaacta cttactctag cttcccggca acaattaata
     2221 gactggatgg aggcggataa agttgcagga ccacttctgc gctcggccct tccggctggc
     2281 tggtttattg ctgataaatc tggagccggt gagcgtgggt ctcgcggtat cattgcagca
     2341 ctggggccag atggtaagcc ctcccgtatc gtagttatct acacgacggg gagtcaggca
     2401 actatggatg aacgaaatag acagatcgct gagataggtg cctcactgat taagcattgg
     2461 ggccctgact acaaagacca tgacggtgat tataaagatc atgacatcga ttacaaggat
     2521 gacgatgaca agggccctca ccaccaccat caccactaat aataaggatc ccccgacctc
     2581 gacctctggc taataaagga aatttatttt cattgcaata gtgtgttgga attttttgtg
     2641 tctctcactc ggaaggacat atgggagggc aaatcatttg gtcgagatcc ctcggagatc
     2701 tctagctaga ggatcgatcc ccgccccgga cgaactaaac ctgactacga catctctgcc
     2761 ccttcttcgc ggggcagtgc atgtaatccc ttcagttggt tggtacaact tgccaactgg
     2821 gccctgttcc acatgtgaca cgggggggga ccaaacacaa aggggttctc tgactgtagt
     2881 tgacatcctt ataaatggat gtgcacattt gccaacactg agtggctttc atcctggagc
     2941 agactttgca gtctgtggac tgcaacacaa cattgccttt atgtgtaact cttggctgaa
     3001 gctcttacac caatgctggg ggacatgtac ctcccagggg cccaggaaga ctacgggagg
     3061 ctacaccaac gtcaatcaga ggggcctgtg tagctaccga taagcggacc ctcaagaggg
     3121 cattagcaat agtgtttata aggccccctt gttaacccta aacgggtagc atatgcttcc
     3181 cgggtagtag tatatactat ccagactaac cctaattcaa tagcatatgt tacccaacgg
     3241 gaagcatatg ctatcgaatt agggttagta aaagggtcct aaggaacagc gatatctccc
     3301 accccatgag ctgtcacggt tttatttaca tggggtcagg attccacgag ggtagtgaac
     3361 cattttagtc acaagggcag tggctgaaga tcaaggagcg ggcagtgaac tctcctgaat
     3421 cttcgcctgc ttcttcattc tccttcgttt agctaataga ataactgctg agttgtgaac
     3481 agtaaggtgt atgtgaggtg ctcgaaaaca aggtttcagg tgacgccccc agaataaaat
     3541 ttggacgggg ggttcagtgg tggcattgtg ctatgacacc aatataaccc tcacaaaccc
     3601 cttgggcaat aaatactagt gtaggaatga aacattctga atatctttaa caatagaaat
     3661 ccatggggtg gggacaagcc gtaaagactg gatgtccatc tcacacgaat ttatggctat
     3721 gggcaacaca taatcctagt gcaatatgat actggggtta ttaagatgtg tcccaggcag
     3781 ggaccaagac aggtgaacca tgttgttaca ctctatttgt aacaagggga aagagagtgg
     3841 acgccgacag cagcggactc cactggttgt ctctaacacc cccgaaaatt aaacggggct
     3901 ccacgccaat ggggcccata aacaaagaca agtggccact cttttttttg aaattgtgga
     3961 gtgggggcac gcgtcagccc ccacacgccg ccctgcggtt ttggactgta aaataagggt
     4021 gtaataactt ggctgattgt aaccccgcta accactgcgg tcaaaccact tgcccacaaa
     4081 accactaatg gcaccccggg gaatacctgc ataagtaggt gggcgggcca agataggggc
     4141 gcgattgctg cgatctggag gacaaattac acacacttgc gcctgagcgc caagcacagg
     4201 gttgttggtc ctcatattca cgaggtcgct gagagcacgg tgggctaatg ttgccatggg
     4261 tagcatatac tacccaaata tctggatagc atatgctatc ctaatctata tctgggtagc
     4321 ataggctatc ctaatctata tctgggtagc atatgctatc ctaatctata tctgggtagt
     4381 atatgctatc ctaatttata tctgggtagc ataggctatc ctaatctata tctgggtagc
     4441 atatgctatc ctaatctata tctgggtagt atatgctatc ctaatctgta tccgggtagc
     4501 atatgctatc ctaatagaga ttagggtagt atatgctatc ctaatttata tctgggtagc
     4561 atatactacc caaatatctg gatagcatat gctatcctaa tctatatctg ggtagcatat
     4621 gctatcctaa tctatatctg ggtagcatag gctatcctaa tctatatctg ggtagcatat
     4681 gctatcctaa tctatatctg ggtagtatat gctatcctaa tttatatctg ggtagcatag
     4741 gctatcctaa tctatatctg ggtagcatat gctatcctaa tctatatctg ggtagtatat
     4801 gctatcctaa tctgtatccg ggtagcatat gctatcctca tgataagctg tcaaacatga
     4861 gaattttctt gaagacgaaa gggcctcgtg atacgcctat ttttataggt taatgtcatg
     4921 ataataatgg tttcttagac gtcaggtggc acttttcggg gaaatgtgcg cggaacccct
     4981 atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga
     5041 taaatgcttc aataatattg aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc
     5101 cttattccct tttttgcggc attttgcctt cctgtttttg ctcacccaga aacgctggtg
     5161 aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg gttacatcga actggatctc
     5221 aacagcggta agatccttga gagttttcgc cccgaagaac gttttccaat gatgagcact
     5281 tttaaagttc tgctatgtgg cgcggtatta tcccgtgttg acgccgggca agagcaactc
     5341 ggtcgccgca tacactattc tcagaatgac ttggttgagt actcaccagt cacagaaaag
     5401 catcttacgg atggcatgac agtaagagaa ttatgcagtg ctgccataac catgagtgat
     5461 aacactgcgg ccaacttact tctgacaacg atcggaggac cgaaggagct aaccgctttt
     5521 ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa
     5581 gccataccaa acgacgagcg tgacaccacg atgcctgcag caatggcaac aacgttgcgc
     5641 aaactattaa ctggcgaact acttactcta gcttcccggc aacaattaat agactggatg
     5701 gaggcggata aagttgcagg accacttctg cgctcggccc ttccggctgg ctggtttatt
     5761 gctgataaat ctggagccgg tgagcgtggg tctcgcggta tcattgcagc actggggcca
     5821 gatggtaagc cctcccgtat cgtagttatc tacacgacgg ggagtcaggc aactatggat
     5881 gaacgaaata gacagatcgc tgagataggt gcctcactga ttaagcattg gtaactgtca
     5941 gaccaagttt actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg
     6001 atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg
     6061 ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt
     6121 ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg
     6181 ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata
     6241 ccaaatactg ttcttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca
     6301 ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag
     6361 tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc
     6421 tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga
     6481 tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg
     6541 tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac
     6601 gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg
     6661 tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg
     6721 ttcctggcct tttgctggcc ttttgctcac atgttctttc ctgcgttatc ccctgattct
     6781 gtggataacc gtattaccgc ctttgagtga gctgataccg ctcgccgcag ccgaacgacc
     6841 gagcgcagcg agtcagtgag cgaggaagcg gaagagcgcc caatacgcaa accgcctctc
     6901 cccgcgcgtt ggccgattca ttaatgcagc tggcacgaca ggtttcccga ctggaaagcg
     6961 ggcagtgagc gcaacgcaat taatgtgagt tagctcactc attaggcacc ccaggcttta
     7021 cactttatgc ttccggctcg tatgttgtgt ggaattgtga gcggataaca atttcacaca
     7081 ggaaacagct atgaccatga ttacgccaag ctctagctag aggtcgacca attctcatgt
     7141 ttgacagctt atcatcgcag atccgggcaa cgttgttgcc attgctgcag gcgcagaact
     7201 ggtaggtatg gaagatctat acattgaatc aatattggca attagccata ttagtcattg
     7261 gttatatagc ataaatcaat attggctatt ggccattgca tacgttgtat ctatatcata
     7321 atatgtacat ttatattggc tcatgtccaa tatgaccgcc atgttgacat tgattattga
     7381 ctagttatta atagtaatca attacggggt cattagttca tagcccatat atggagttcc
     7441 gcgttacata acttacggta aatggcccgc ctggctgacc gcccaacgac ccccgcccat
     7501 tgacgtcaat aatgacgtat gttcccatag taacgccaat agggactttc cattgacgtc
     7561 aatgggtgga gtatttacgg taaactgccc acttggcagt acatcaagtg tatcatatgc
     7621 caagtccgcc ccctattgac gtcaatgacg gtaaatggcc cgcctggcat tatgcccagt
     7681 acatgacctt acgggacttt cctacttggc agtacatcta cgtattagtc atcgctatta
     7741 ccatggtgat gcggttttgg cagtacacca atgggcgtgg atagcggttt gactcacggg
     7801 gatttccaag tctccacccc attgacgtca atgggagttt gttttggcac caaaatcaac
     7861 gggactttcc aaaatgtcgt aataaccccg ccccgttgac gcaaatgggc ggtaggcgtg
     7921 tacggtggga ggtctatata agcagagctc gtttagtgaa ccgtcagatc ctcactctct
     7981 tccgcatcgc tgtctgcgag ggccagctgt tgggctcgcg gttgaggaca aactcttcgc
     8041 ggtctttcca gtactcttgg atcggaaacc cgtcggcctc cgaacggtac tccgccaccg
     8101 agggacctga gcgagtccgc atcgaccgga tcggaaaacc t
//
