LOCUS       sequence-208722-        8461 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 8461)
  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 8461)
  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: WARP-COMP-blac-flag-his
FEATURES             Location/Qualifiers
     source          1..8461
                     /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             1993..2778
                     /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             2785..2850
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             2857..2874
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    2912..2967
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(2966..2985)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(5203..5221)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5289..5393
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5394..5462,5463..6254)
                     /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      6425..7013
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6914..6933
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7167..7184
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    7301..7322
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7337..7342,7343..7360,7361..7367)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7375..7391
                     /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     7380..7402
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7399..7415
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7399..7415
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     enhancer        7686..8065
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        8066..8269
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     8220..8240
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     8266..8290
                     /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 atgaacgtgg tgtcccacaa cagcatgaag aaagtgggcg aggaaggcaa ggacgagaag
      481 atcatcagct gcgtggtgga atacgtgatc aagggcgacc tgagcatcga cgacggcgat
      541 ttctgcgaca gcaacagcat ctgcagcaac aaccccggcc tgtgcgacaa cggccaggtg
      601 gtggtgcccg gcgactactg ctccaactac tacgacatca ccctggtggt ggaagagtcc
      661 agcttcgtgc agaaggacta ctggctgaag ggcaccatcc cctttctgga atccatggtg
      721 ttcaacagca acgtggccaa ggacaaggcc cacatgagcg tggtgctgtt cgccggcaag
      781 cagcgggtgg tggtcccctt caccgacgag agcagccagg acaagaacat gctgctggaa
      841 aagatcaact ccatcgacga gggaatcgcc accagcagcg acaccctgta cgtgaacgcc
      901 ctgaagtacg ccatcgagaa cgtgatcttc ggcgacggca cccggaagga cgcccctaag
      961 gtggccgtgc tgttctacta cggcttcgac tacggcgcca acaagaatat gatccccgac
     1021 gtggtgcaga cctacaaaga acagaacgtg aagctgatca tcgtggggat cgccctgggc
     1081 atccgggaca acgcctacct gctggccgac tgcagcatcg gcgacgagaa ctgtgtgaat
     1141 gtggtgttta agccctggga cttcgtgatc cccgctgccg ccgacgtgaa ccagaagatc
     1201 tgcgataaga acaaggccgg cgcgccgtcg acctccatca cggcctataa gagtgagggg
     1261 gagtcagcgg agttctcctt cccactcaac cttggagagg aaagcctgca gggagagttg
     1321 agatggaagg cagagaaggc tccttcttcc cagtcctgga tcaccttctc cctaaagaac
     1381 caaaaggtgt ctgtgcagaa gtctactagc aaccccaagt tccagctgtc cgaaacgctc
     1441 ccactcaccc ttcagatacc ccaggtctcc cttcagtttg ctggttctgg caacctgacc
     1501 ctgactctgg acagagggat actgtatcag gaagtgaacc tggtggtgat gaaagtgact
     1561 cagcccgaca gcaacacttt gacctgtgag gtgatgggac ccacctcacc caagatgaga
     1621 ctgatcttga agcaggagaa tcaggaggcc agggtctcca ggcaggagaa agtgattcaa
     1681 gtgcaggccc ctgaagcagg ggtgtggcaa tgtctactga gtgaaggtga agaggtcaag
     1741 atggactcca agatccaggt tttatccaaa gggttgaatt ccggcggcgg ctccggaggc
     1801 ggtaccggtg gcggcgacct agccccacag atgcttcgag aactccagga gactaatgcg
     1861 gcgctgcaag acgtgagaga gctcttgcga cagcaggtca aggagatcac cttcctgaag
     1921 aatacggtga tggaatgtga cgcctgcggt ctagacagaa acctgcctcc actagcacct
     1981 ctgggcccac acccagaaac gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca
     2041 cgagtgggtt acatcgaact ggatctcaac agcggtaaga tccttgagag ttttcgcccc
     2101 gaagaacgtt ttccaatgat gagcactttt aaagttctgc tatgtggcgc ggtattatcc
     2161 cgtattgacg ccgggcaaga gcaactcggt cgccgcatac actattctca gaatgacttg
     2221 gttgagtact caccagtcac agaaaagcat cttacggatg gcatgacagt aagagaatta
     2281 tgcagtgctg ccataaccat gagtgataac actgcggcca acttacttct gacaacgatc
     2341 ggaggaccga aggagctaac cgcttttttg cacaacatgg gggatcatgt aactcgcctt
     2401 gatcgttggg aaccggagct gaatgaagcc ataccaaacg acgagcgtga caccacgatg
     2461 cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg gcgaactact tactctagct
     2521 tcccggcaac aattaataga ctggatggag gcggataaag ttgcaggacc acttctgcgc
     2581 tcggcccttc cggctggctg gtttattgct gataaatctg gagccggtga gcgtgggtct
     2641 cgcggtatca ttgcagcact ggggccagat ggtaagccct cccgtatcgt agttatctac
     2701 acgacgggga gtcaggcaac tatggatgaa cgaaatagac agatcgctga gataggtgcc
     2761 tcactgatta agcattgggg ccctgactac aaagaccatg acggtgatta taaagatcat
     2821 gacatcgatt acaaggatga cgatgacaag ggccctcacc accaccatca ccactaatta
     2881 attaaggatc ccccgacctc gacctctggc taataaagga aatttatttt cattgcaata
     2941 gtgtgttgga attttttgtg tctctcactc ggaaggacat atgggagggc aaatcatttg
     3001 gtcgagatcc ctcggagatc tctagctaga ggatcgatcc ccgccccgga cgaactaaac
     3061 ctgactacga catctctgcc ccttcttcgc ggggcagtgc atgtaatccc ttcagttggt
     3121 tggtacaact tgccaactgg gccctgttcc acatgtgaca cgggggggga ccaaacacaa
     3181 aggggttctc tgactgtagt tgacatcctt ataaatggat gtgcacattt gccaacactg
     3241 agtggctttc atcctggagc agactttgca gtctgtggac tgcaacacaa cattgccttt
     3301 atgtgtaact cttggctgaa gctcttacac caatgctggg ggacatgtac ctcccagggg
     3361 cccaggaaga ctacgggagg ctacaccaac gtcaatcaga ggggcctgtg tagctaccga
     3421 taagcggacc ctcaagaggg cattagcaat agtgtttata aggccccctt gttaacccta
     3481 aacgggtagc atatgcttcc cgggtagtag tatatactat ccagactaac cctaattcaa
     3541 tagcatatgt tacccaacgg gaagcatatg ctatcgaatt agggttagta aaagggtcct
     3601 aaggaacagc gatatctccc accccatgag ctgtcacggt tttatttaca tggggtcagg
     3661 attccacgag ggtagtgaac cattttagtc acaagggcag tggctgaaga tcaaggagcg
     3721 ggcagtgaac tctcctgaat cttcgcctgc ttcttcattc tccttcgttt agctaataga
     3781 ataactgctg agttgtgaac agtaaggtgt atgtgaggtg ctcgaaaaca aggtttcagg
     3841 tgacgccccc agaataaaat ttggacgggg ggttcagtgg tggcattgtg ctatgacacc
     3901 aatataaccc tcacaaaccc cttgggcaat aaatactagt gtaggaatga aacattctga
     3961 atatctttaa caatagaaat ccatggggtg gggacaagcc gtaaagactg gatgtccatc
     4021 tcacacgaat ttatggctat gggcaacaca taatcctagt gcaatatgat actggggtta
     4081 ttaagatgtg tcccaggcag ggaccaagac aggtgaacca tgttgttaca ctctatttgt
     4141 aacaagggga aagagagtgg acgccgacag cagcggactc cactggttgt ctctaacacc
     4201 cccgaaaatt aaacggggct ccacgccaat ggggcccata aacaaagaca agtggccact
     4261 cttttttttg aaattgtgga gtgggggcac gcgtcagccc ccacacgccg ccctgcggtt
     4321 ttggactgta aaataagggt gtaataactt ggctgattgt aaccccgcta accactgcgg
     4381 tcaaaccact tgcccacaaa accactaatg gcaccccggg gaatacctgc ataagtaggt
     4441 gggcgggcca agataggggc gcgattgctg cgatctggag gacaaattac acacacttgc
     4501 gcctgagcgc caagcacagg gttgttggtc ctcatattca cgaggtcgct gagagcacgg
     4561 tgggctaatg ttgccatggg tagcatatac tacccaaata tctggatagc atatgctatc
     4621 ctaatctata tctgggtagc ataggctatc ctaatctata tctgggtagc atatgctatc
     4681 ctaatctata tctgggtagt atatgctatc ctaatttata tctgggtagc ataggctatc
     4741 ctaatctata tctgggtagc atatgctatc ctaatctata tctgggtagt atatgctatc
     4801 ctaatctgta tccgggtagc atatgctatc ctaatagaga ttagggtagt atatgctatc
     4861 ctaatttata tctgggtagc atatactacc caaatatctg gatagcatat gctatcctaa
     4921 tctatatctg ggtagcatat gctatcctaa tctatatctg ggtagcatag gctatcctaa
     4981 tctatatctg ggtagcatat gctatcctaa tctatatctg ggtagtatat gctatcctaa
     5041 tttatatctg ggtagcatag gctatcctaa tctatatctg ggtagcatat gctatcctaa
     5101 tctatatctg ggtagtatat gctatcctaa tctgtatccg ggtagcatat gctatcctca
     5161 tgataagctg tcaaacatga gaattttctt gaagacgaaa gggcctcgtg atacgcctat
     5221 ttttataggt taatgtcatg ataataatgg tttcttagac gtcaggtggc acttttcggg
     5281 gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc
     5341 tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag agtatgagta
     5401 ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg
     5461 ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg
     5521 gttacatcga actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac
     5581 gttttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtgttg
     5641 acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt
     5701 actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg
     5761 ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg atcggaggac
     5821 cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt
     5881 gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgcag
     5941 caatggcaac aacgttgcgc aaactattaa ctggcgaact acttactcta gcttcccggc
     6001 aacaattaat agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc
     6061 ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggg tctcgcggta
     6121 tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg
     6181 ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga
     6241 ttaagcattg gtaactgtca gaccaagttt actcatatat actttagatt gatttaaaac
     6301 ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa
     6361 tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat
     6421 cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc
     6481 taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg
     6541 gcttcagcag agcgcagata ccaaatactg ttcttctagt gtagccgtag ttaggccacc
     6601 acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg
     6661 ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg
     6721 ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa
     6781 cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg
     6841 aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga
     6901 gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct
     6961 gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca
     7021 gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgttctttc
     7081 ctgcgttatc ccctgattct gtggataacc gtattaccgc ctttgagtga gctgataccg
     7141 ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg gaagagcgcc
     7201 caatacgcaa accgcctctc cccgcgcgtt ggccgattca ttaatgcagc tggcacgaca
     7261 ggtttcccga ctggaaagcg ggcagtgagc gcaacgcaat taatgtgagt tagctcactc
     7321 attaggcacc ccaggcttta cactttatgc ttccggctcg tatgttgtgt ggaattgtga
     7381 gcggataaca atttcacaca ggaaacagct atgaccatga ttacgccaag ctctagctag
     7441 aggtcgacca attctcatgt ttgacagctt atcatcgcag atccgggcaa cgttgttgcc
     7501 attgctgcag gcgcagaact ggtaggtatg gaagatctat acattgaatc aatattggca
     7561 attagccata ttagtcattg gttatatagc ataaatcaat attggctatt ggccattgca
     7621 tacgttgtat ctatatcata atatgtacat ttatattggc tcatgtccaa tatgaccgcc
     7681 atgttgacat tgattattga ctagttatta atagtaatca attacggggt cattagttca
     7741 tagcccatat atggagttcc gcgttacata acttacggta aatggcccgc ctggctgacc
     7801 gcccaacgac ccccgcccat tgacgtcaat aatgacgtat gttcccatag taacgccaat
     7861 agggactttc cattgacgtc aatgggtgga gtatttacgg taaactgccc acttggcagt
     7921 acatcaagtg tatcatatgc caagtccgcc ccctattgac gtcaatgacg gtaaatggcc
     7981 cgcctggcat tatgcccagt acatgacctt acgggacttt cctacttggc agtacatcta
     8041 cgtattagtc atcgctatta ccatggtgat gcggttttgg cagtacacca atgggcgtgg
     8101 atagcggttt gactcacggg gatttccaag tctccacccc attgacgtca atgggagttt
     8161 gttttggcac caaaatcaac gggactttcc aaaatgtcgt aataaccccg ccccgttgac
     8221 gcaaatgggc ggtaggcgtg tacggtggga ggtctatata agcagagctc gtttagtgaa
     8281 ccgtcagatc ctcactctct tccgcatcgc tgtctgcgag ggccagctgt tgggctcgcg
     8341 gttgaggaca aactcttcgc ggtctttcca gtactcttgg atcggaaacc cgtcggcctc
     8401 cgaacggtac tccgccaccg agggacctga gcgagtccgc atcgaccgga tcggaaaacc
     8461 t
//
