LOCUS       sequence-208690-        8938 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 8938)
  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 8938)
  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_1463900-COMP-blac-flag-his
FEATURES             Location/Qualifiers
     source          1..8938
                     /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             2470..3255
                     /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             3262..3327
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             3334..3351
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    3389..3444
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(3443..3462)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(5680..5698)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5766..5870
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5871..5939,5940..6731)
                     /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      6902..7490
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7391..7410
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7644..7661
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    7778..7799
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7814..7819,7820..7837,7838..7844)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7852..7868
                     /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     7857..7879
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7876..7892
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7876..7892
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     enhancer        8163..8542
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        8543..8746
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     8697..8717
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     8743..8767
                     /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 ggccgccaac
      421 gagaacaact tcttcatcaa gatcctgcgg agccaggaaa agaagatcga gaacgaagtc
      481 tccctgtacg agaagctgaa caaccggatc ctgttcgagg gcagcgacga cttcagcaac
      541 ctggacaagc tgaagtccct gctgagcttc cagaagaaga ttgacaacac catccgcaag
      601 aagcagaaag aggaaaacaa gctcaagaag ttccagcagg ccatcaagaa cggcaacaac
      661 atctaccccg tgggcaacaa ccctcagggc ttcgagaaga aagagcgcga ggaaaaagag
      721 caggaaaaaa cctacatgaa caacgtgatc aacaagatca acgagcagaa gggcaactac
      781 tacgagccca acaagaacat caacaacaag tacaccgagg acctgaccag caacggcctg
      841 ctgtacaaca aggataagat caagaaccag ttcaacaatc cccaccagga aaaattcatc
      901 cggaagcacg agcacgagag agagcacgag ggcaagaacg aaggcaaaaa cgagcacaag
      961 aatgagcata agaacgaaca caaaaatgaa cacgtccacg tgcacgagca taacagcaag
     1021 aactatgagg gcaaccctcc caagaacgag taccccaaca tcaacttcgc caacatcctg
     1081 aacagcatcg acttctacaa gaaaatcgac ctggtcaaga actacttcgg ctacaacaac
     1141 aacgaggccc tgaaccagaa catgaaggaa cagaaaaaca tgcagaacca ccacgtggaa
     1201 aacgagaaga acaacatgat ccaggccaac aaccaggtgg acgagaatca gaacaatcac
     1261 cagtataaca acaacttcca caaccagaag aacgacaacc tgaagcagga caccaaggac
     1321 aacagcgccg agcagaccta caacaatgcc aacaacaata tcgaggacaa caatcacatc
     1381 aagttcttca tctccaagga catcgagaaa aaaaagaaca agcagaacga catcaatgag
     1441 gccgtgtaca acgccatcaa aaacaagaac ctgcacctcc ggcggaacaa catcaaagaa
     1501 atcttcgacg tggacgacct cgtgaagaat atcaagagct tcctgggcat caagagcaac
     1561 atccacgagg ccttcgagaa ccagaagctg atcatcaaga attgcaacta cgagagcttc
     1621 ggccccgagc tgtgctccgt ggatgagaag gccaaagaaa tgctgtggaa gtatgagaag
     1681 aagaaaaact ccgccggcgc gccgtcgacc tccatcacgg cctataagag tgagggggag
     1741 tcagcggagt tctccttccc actcaacctt ggagaggaaa gcctgcaggg agagttgaga
     1801 tggaaggcag agaaggctcc ttcttcccag tcctggatca ccttctccct aaagaaccaa
     1861 aaggtgtctg tgcagaagtc tactagcaac cccaagttcc agctgtccga aacgctccca
     1921 ctcacccttc agatacccca ggtctccctt cagtttgctg gttctggcaa cctgaccctg
     1981 actctggaca gagggatact gtatcaggaa gtgaacctgg tggtgatgaa agtgactcag
     2041 cccgacagca acactttgac ctgtgaggtg atgggaccca cctcacccaa gatgagactg
     2101 atcttgaagc aggagaatca ggaggccagg gtctccaggc aggagaaagt gattcaagtg
     2161 caggcccctg aagcaggggt gtggcaatgt ctactgagtg aaggtgaaga ggtcaagatg
     2221 gactccaaga tccaggtttt atccaaaggg ttgaattccg gcggcggctc cggaggcggt
     2281 accggtggcg gcgacctagc cccacagatg cttcgagaac tccaggagac taatgcggcg
     2341 ctgcaagacg tgagagagct cttgcgacag caggtcaagg agatcacctt cctgaagaat
     2401 acggtgatgg aatgtgacgc ctgcggtcta gacagaaacc tgcctccact agcacctctg
     2461 ggcccacacc cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcacga
     2521 gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa
     2581 gaacgttttc caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt
     2641 attgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt
     2701 gagtactcac cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc
     2761 agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac aacgatcgga
     2821 ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat
     2881 cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct
     2941 gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc
     3001 cggcaacaat taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg
     3061 gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc
     3121 ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg
     3181 acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca
     3241 ctgattaagc attggggccc tgactacaaa gaccatgacg gtgattataa agatcatgac
     3301 atcgattaca aggatgacga tgacaagggc cctcaccacc accatcacca ctaattaatt
     3361 aaggatcccc cgacctcgac ctctggctaa taaaggaaat ttattttcat tgcaatagtg
     3421 tgttggaatt ttttgtgtct ctcactcgga aggacatatg ggagggcaaa tcatttggtc
     3481 gagatccctc ggagatctct agctagagga tcgatccccg ccccggacga actaaacctg
     3541 actacgacat ctctgcccct tcttcgcggg gcagtgcatg taatcccttc agttggttgg
     3601 tacaacttgc caactgggcc ctgttccaca tgtgacacgg ggggggacca aacacaaagg
     3661 ggttctctga ctgtagttga catccttata aatggatgtg cacatttgcc aacactgagt
     3721 ggctttcatc ctggagcaga ctttgcagtc tgtggactgc aacacaacat tgcctttatg
     3781 tgtaactctt ggctgaagct cttacaccaa tgctggggga catgtacctc ccaggggccc
     3841 aggaagacta cgggaggcta caccaacgtc aatcagaggg gcctgtgtag ctaccgataa
     3901 gcggaccctc aagagggcat tagcaatagt gtttataagg cccccttgtt aaccctaaac
     3961 gggtagcata tgcttcccgg gtagtagtat atactatcca gactaaccct aattcaatag
     4021 catatgttac ccaacgggaa gcatatgcta tcgaattagg gttagtaaaa gggtcctaag
     4081 gaacagcgat atctcccacc ccatgagctg tcacggtttt atttacatgg ggtcaggatt
     4141 ccacgagggt agtgaaccat tttagtcaca agggcagtgg ctgaagatca aggagcgggc
     4201 agtgaactct cctgaatctt cgcctgcttc ttcattctcc ttcgtttagc taatagaata
     4261 actgctgagt tgtgaacagt aaggtgtatg tgaggtgctc gaaaacaagg tttcaggtga
     4321 cgcccccaga ataaaatttg gacggggggt tcagtggtgg cattgtgcta tgacaccaat
     4381 ataaccctca caaacccctt gggcaataaa tactagtgta ggaatgaaac attctgaata
     4441 tctttaacaa tagaaatcca tggggtgggg acaagccgta aagactggat gtccatctca
     4501 cacgaattta tggctatggg caacacataa tcctagtgca atatgatact ggggttatta
     4561 agatgtgtcc caggcaggga ccaagacagg tgaaccatgt tgttacactc tatttgtaac
     4621 aaggggaaag agagtggacg ccgacagcag cggactccac tggttgtctc taacaccccc
     4681 gaaaattaaa cggggctcca cgccaatggg gcccataaac aaagacaagt ggccactctt
     4741 ttttttgaaa ttgtggagtg ggggcacgcg tcagccccca cacgccgccc tgcggttttg
     4801 gactgtaaaa taagggtgta ataacttggc tgattgtaac cccgctaacc actgcggtca
     4861 aaccacttgc ccacaaaacc actaatggca ccccggggaa tacctgcata agtaggtggg
     4921 cgggccaaga taggggcgcg attgctgcga tctggaggac aaattacaca cacttgcgcc
     4981 tgagcgccaa gcacagggtt gttggtcctc atattcacga ggtcgctgag agcacggtgg
     5041 gctaatgttg ccatgggtag catatactac ccaaatatct ggatagcata tgctatccta
     5101 atctatatct gggtagcata ggctatccta atctatatct gggtagcata tgctatccta
     5161 atctatatct gggtagtata tgctatccta atttatatct gggtagcata ggctatccta
     5221 atctatatct gggtagcata tgctatccta atctatatct gggtagtata tgctatccta
     5281 atctgtatcc gggtagcata tgctatccta atagagatta gggtagtata tgctatccta
     5341 atttatatct gggtagcata tactacccaa atatctggat agcatatgct atcctaatct
     5401 atatctgggt agcatatgct atcctaatct atatctgggt agcataggct atcctaatct
     5461 atatctgggt agcatatgct atcctaatct atatctgggt agtatatgct atcctaattt
     5521 atatctgggt agcataggct atcctaatct atatctgggt agcatatgct atcctaatct
     5581 atatctgggt agtatatgct atcctaatct gtatccgggt agcatatgct atcctcatga
     5641 taagctgtca aacatgagaa ttttcttgaa gacgaaaggg cctcgtgata cgcctatttt
     5701 tataggttaa tgtcatgata ataatggttt cttagacgtc aggtggcact tttcggggaa
     5761 atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca
     5821 tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt atgagtattc
     5881 aacatttccg tgtcgccctt attccctttt ttgcggcatt ttgccttcct gtttttgctc
     5941 acccagaaac gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt
     6001 acatcgaact ggatctcaac agcggtaaga tccttgagag ttttcgcccc gaagaacgtt
     6061 ttccaatgat gagcactttt aaagttctgc tatgtggcgc ggtattatcc cgtgttgacg
     6121 ccgggcaaga gcaactcggt cgccgcatac actattctca gaatgacttg gttgagtact
     6181 caccagtcac agaaaagcat cttacggatg gcatgacagt aagagaatta tgcagtgctg
     6241 ccataaccat gagtgataac actgcggcca acttacttct gacaacgatc ggaggaccga
     6301 aggagctaac cgcttttttg cacaacatgg gggatcatgt aactcgcctt gatcgttggg
     6361 aaccggagct gaatgaagcc ataccaaacg acgagcgtga caccacgatg cctgcagcaa
     6421 tggcaacaac gttgcgcaaa ctattaactg gcgaactact tactctagct tcccggcaac
     6481 aattaataga ctggatggag gcggataaag ttgcaggacc acttctgcgc tcggcccttc
     6541 cggctggctg gtttattgct gataaatctg gagccggtga gcgtgggtct cgcggtatca
     6601 ttgcagcact ggggccagat ggtaagccct cccgtatcgt agttatctac acgacgggga
     6661 gtcaggcaac tatggatgaa cgaaatagac agatcgctga gataggtgcc tcactgatta
     6721 agcattggta actgtcagac caagtttact catatatact ttagattgat ttaaaacttc
     6781 atttttaatt taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc
     6841 cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt
     6901 cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac
     6961 cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct
     7021 tcagcagagc gcagatacca aatactgttc ttctagtgta gccgtagtta ggccaccact
     7081 tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg
     7141 ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata
     7201 aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga
     7261 cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag
     7321 ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg
     7381 agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac
     7441 ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca
     7501 acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg ttctttcctg
     7561 cgttatcccc tgattctgtg gataaccgta ttaccgcctt tgagtgagct gataccgctc
     7621 gccgcagccg aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa gagcgcccaa
     7681 tacgcaaacc gcctctcccc gcgcgttggc cgattcatta atgcagctgg cacgacaggt
     7741 ttcccgactg gaaagcgggc agtgagcgca acgcaattaa tgtgagttag ctcactcatt
     7801 aggcacccca ggctttacac tttatgcttc cggctcgtat gttgtgtgga attgtgagcg
     7861 gataacaatt tcacacagga aacagctatg accatgatta cgccaagctc tagctagagg
     7921 tcgaccaatt ctcatgtttg acagcttatc atcgcagatc cgggcaacgt tgttgccatt
     7981 gctgcaggcg cagaactggt aggtatggaa gatctataca ttgaatcaat attggcaatt
     8041 agccatatta gtcattggtt atatagcata aatcaatatt ggctattggc cattgcatac
     8101 gttgtatcta tatcataata tgtacattta tattggctca tgtccaatat gaccgccatg
     8161 ttgacattga ttattgacta gttattaata gtaatcaatt acggggtcat tagttcatag
     8221 cccatatatg gagttccgcg ttacataact tacggtaaat ggcccgcctg gctgaccgcc
     8281 caacgacccc cgcccattga cgtcaataat gacgtatgtt cccatagtaa cgccaatagg
     8341 gactttccat tgacgtcaat gggtggagta tttacggtaa actgcccact tggcagtaca
     8401 tcaagtgtat catatgccaa gtccgccccc tattgacgtc aatgacggta aatggcccgc
     8461 ctggcattat gcccagtaca tgaccttacg ggactttcct acttggcagt acatctacgt
     8521 attagtcatc gctattacca tggtgatgcg gttttggcag tacaccaatg ggcgtggata
     8581 gcggtttgac tcacggggat ttccaagtct ccaccccatt gacgtcaatg ggagtttgtt
     8641 ttggcaccaa aatcaacggg actttccaaa atgtcgtaat aaccccgccc cgttgacgca
     8701 aatgggcggt aggcgtgtac ggtgggaggt ctatataagc agagctcgtt tagtgaaccg
     8761 tcagatcctc actctcttcc gcatcgctgt ctgcgagggc cagctgttgg gctcgcggtt
     8821 gaggacaaac tcttcgcggt ctttccagta ctcttggatc ggaaacccgt cggcctccga
     8881 acggtactcc gccaccgagg gacctgagcg agtccgcatc gaccggatcg gaaaacct
//
