LOCUS       sequence-208700-        8578 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 8578)
  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 8578)
  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_0511400-COMP-blac-flag-his
FEATURES             Location/Qualifiers
     source          1..8578
                     /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             2110..2895
                     /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             2902..2967
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             2974..2991
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    3029..3084
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(3083..3102)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(5320..5338)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5406..5510
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5511..5579,5580..6371)
                     /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      6542..7130
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7031..7050
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7284..7301
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    7418..7439
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7454..7459,7460..7477,7478..7484)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7492..7508
                     /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     7497..7519
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7516..7532
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7516..7532
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     enhancer        7803..8182
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        8183..8386
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     8337..8357
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     8383..8407
                     /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 ggccgccagc
      421 gacaccttcc gggtgttcga ggacttcgac gtgcggcgga agaacaagat ctggtgctgc
      481 aacctgaacc acgagaagtc ctacttccac aacaaggcca tcaacaccgc ctacaagaac
      541 cgggtgctga aagagagcat cctgctgaac ctggacctgg acatgaagaa gtacaaggac
      601 aacatcatca cccggaaaaa gacccctgag aacatctaca aagagatcta cgagaacaac
      661 tacgagatga agtacgacga ggacatcccc aacaacatgg ccgaggaaaa gcgcgacgag
      721 aaagaagtga tcgagcacct ggaaatcgac gagaagaacg gcaagcagta caagagcgac
      781 atcaacaagc ccgtgtccct gagccacctg aagcagtata agaatatcta tgtgaacaac
      841 aacaataaga tcaacaagaa gaagtccatc gacaagcacc tccccagcta caacctggaa
      901 cgcaagaaca acaagtacct gaacttcctg ctggtggaca accggaacga ggcctacacc
      961 ttcatggtgc ctatgaagtt ctacatcaat cacgagatgt acaatatcgc cgacgaagag
     1021 tacaacaagc tgatggaaga taacagcgtg gacgtctacc tgaacaacat cctggtcgag
     1081 tataagtacg agaacttcga gatcaaagag ggcgaggtcg acggggaggt ggaaggcgag
     1141 ggcgaagtgg aaggtgaagt cgagggagag gtcaaaggca aggtggtgga agggatcgag
     1201 aacaatatga acgaggaaga gaagtataac aaggataaca aagacaaaga gaaccagatc
     1261 aacagcaacg gccaggacga gaacaccgag ttccaggaaa acgacaacaa cgacgccgtg
     1321 atcatgaagt acaccggcgc gccgtcgacc tccatcacgg cctataagag tgagggggag
     1381 tcagcggagt tctccttccc actcaacctt ggagaggaaa gcctgcaggg agagttgaga
     1441 tggaaggcag agaaggctcc ttcttcccag tcctggatca ccttctccct aaagaaccaa
     1501 aaggtgtctg tgcagaagtc tactagcaac cccaagttcc agctgtccga aacgctccca
     1561 ctcacccttc agatacccca ggtctccctt cagtttgctg gttctggcaa cctgaccctg
     1621 actctggaca gagggatact gtatcaggaa gtgaacctgg tggtgatgaa agtgactcag
     1681 cccgacagca acactttgac ctgtgaggtg atgggaccca cctcacccaa gatgagactg
     1741 atcttgaagc aggagaatca ggaggccagg gtctccaggc aggagaaagt gattcaagtg
     1801 caggcccctg aagcaggggt gtggcaatgt ctactgagtg aaggtgaaga ggtcaagatg
     1861 gactccaaga tccaggtttt atccaaaggg ttgaattccg gcggcggctc cggaggcggt
     1921 accggtggcg gcgacctagc cccacagatg cttcgagaac tccaggagac taatgcggcg
     1981 ctgcaagacg tgagagagct cttgcgacag caggtcaagg agatcacctt cctgaagaat
     2041 acggtgatgg aatgtgacgc ctgcggtcta gacagaaacc tgcctccact agcacctctg
     2101 ggcccacacc cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcacga
     2161 gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa
     2221 gaacgttttc caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt
     2281 attgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt
     2341 gagtactcac cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc
     2401 agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac aacgatcgga
     2461 ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat
     2521 cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct
     2581 gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc
     2641 cggcaacaat taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg
     2701 gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc
     2761 ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg
     2821 acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca
     2881 ctgattaagc attggggccc tgactacaaa gaccatgacg gtgattataa agatcatgac
     2941 atcgattaca aggatgacga tgacaagggc cctcaccacc accatcacca ctaattaatt
     3001 aaggatcccc cgacctcgac ctctggctaa taaaggaaat ttattttcat tgcaatagtg
     3061 tgttggaatt ttttgtgtct ctcactcgga aggacatatg ggagggcaaa tcatttggtc
     3121 gagatccctc ggagatctct agctagagga tcgatccccg ccccggacga actaaacctg
     3181 actacgacat ctctgcccct tcttcgcggg gcagtgcatg taatcccttc agttggttgg
     3241 tacaacttgc caactgggcc ctgttccaca tgtgacacgg ggggggacca aacacaaagg
     3301 ggttctctga ctgtagttga catccttata aatggatgtg cacatttgcc aacactgagt
     3361 ggctttcatc ctggagcaga ctttgcagtc tgtggactgc aacacaacat tgcctttatg
     3421 tgtaactctt ggctgaagct cttacaccaa tgctggggga catgtacctc ccaggggccc
     3481 aggaagacta cgggaggcta caccaacgtc aatcagaggg gcctgtgtag ctaccgataa
     3541 gcggaccctc aagagggcat tagcaatagt gtttataagg cccccttgtt aaccctaaac
     3601 gggtagcata tgcttcccgg gtagtagtat atactatcca gactaaccct aattcaatag
     3661 catatgttac ccaacgggaa gcatatgcta tcgaattagg gttagtaaaa gggtcctaag
     3721 gaacagcgat atctcccacc ccatgagctg tcacggtttt atttacatgg ggtcaggatt
     3781 ccacgagggt agtgaaccat tttagtcaca agggcagtgg ctgaagatca aggagcgggc
     3841 agtgaactct cctgaatctt cgcctgcttc ttcattctcc ttcgtttagc taatagaata
     3901 actgctgagt tgtgaacagt aaggtgtatg tgaggtgctc gaaaacaagg tttcaggtga
     3961 cgcccccaga ataaaatttg gacggggggt tcagtggtgg cattgtgcta tgacaccaat
     4021 ataaccctca caaacccctt gggcaataaa tactagtgta ggaatgaaac attctgaata
     4081 tctttaacaa tagaaatcca tggggtgggg acaagccgta aagactggat gtccatctca
     4141 cacgaattta tggctatggg caacacataa tcctagtgca atatgatact ggggttatta
     4201 agatgtgtcc caggcaggga ccaagacagg tgaaccatgt tgttacactc tatttgtaac
     4261 aaggggaaag agagtggacg ccgacagcag cggactccac tggttgtctc taacaccccc
     4321 gaaaattaaa cggggctcca cgccaatggg gcccataaac aaagacaagt ggccactctt
     4381 ttttttgaaa ttgtggagtg ggggcacgcg tcagccccca cacgccgccc tgcggttttg
     4441 gactgtaaaa taagggtgta ataacttggc tgattgtaac cccgctaacc actgcggtca
     4501 aaccacttgc ccacaaaacc actaatggca ccccggggaa tacctgcata agtaggtggg
     4561 cgggccaaga taggggcgcg attgctgcga tctggaggac aaattacaca cacttgcgcc
     4621 tgagcgccaa gcacagggtt gttggtcctc atattcacga ggtcgctgag agcacggtgg
     4681 gctaatgttg ccatgggtag catatactac ccaaatatct ggatagcata tgctatccta
     4741 atctatatct gggtagcata ggctatccta atctatatct gggtagcata tgctatccta
     4801 atctatatct gggtagtata tgctatccta atttatatct gggtagcata ggctatccta
     4861 atctatatct gggtagcata tgctatccta atctatatct gggtagtata tgctatccta
     4921 atctgtatcc gggtagcata tgctatccta atagagatta gggtagtata tgctatccta
     4981 atttatatct gggtagcata tactacccaa atatctggat agcatatgct atcctaatct
     5041 atatctgggt agcatatgct atcctaatct atatctgggt agcataggct atcctaatct
     5101 atatctgggt agcatatgct atcctaatct atatctgggt agtatatgct atcctaattt
     5161 atatctgggt agcataggct atcctaatct atatctgggt agcatatgct atcctaatct
     5221 atatctgggt agtatatgct atcctaatct gtatccgggt agcatatgct atcctcatga
     5281 taagctgtca aacatgagaa ttttcttgaa gacgaaaggg cctcgtgata cgcctatttt
     5341 tataggttaa tgtcatgata ataatggttt cttagacgtc aggtggcact tttcggggaa
     5401 atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca
     5461 tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt atgagtattc
     5521 aacatttccg tgtcgccctt attccctttt ttgcggcatt ttgccttcct gtttttgctc
     5581 acccagaaac gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt
     5641 acatcgaact ggatctcaac agcggtaaga tccttgagag ttttcgcccc gaagaacgtt
     5701 ttccaatgat gagcactttt aaagttctgc tatgtggcgc ggtattatcc cgtgttgacg
     5761 ccgggcaaga gcaactcggt cgccgcatac actattctca gaatgacttg gttgagtact
     5821 caccagtcac agaaaagcat cttacggatg gcatgacagt aagagaatta tgcagtgctg
     5881 ccataaccat gagtgataac actgcggcca acttacttct gacaacgatc ggaggaccga
     5941 aggagctaac cgcttttttg cacaacatgg gggatcatgt aactcgcctt gatcgttggg
     6001 aaccggagct gaatgaagcc ataccaaacg acgagcgtga caccacgatg cctgcagcaa
     6061 tggcaacaac gttgcgcaaa ctattaactg gcgaactact tactctagct tcccggcaac
     6121 aattaataga ctggatggag gcggataaag ttgcaggacc acttctgcgc tcggcccttc
     6181 cggctggctg gtttattgct gataaatctg gagccggtga gcgtgggtct cgcggtatca
     6241 ttgcagcact ggggccagat ggtaagccct cccgtatcgt agttatctac acgacgggga
     6301 gtcaggcaac tatggatgaa cgaaatagac agatcgctga gataggtgcc tcactgatta
     6361 agcattggta actgtcagac caagtttact catatatact ttagattgat ttaaaacttc
     6421 atttttaatt taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc
     6481 cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt
     6541 cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac
     6601 cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct
     6661 tcagcagagc gcagatacca aatactgttc ttctagtgta gccgtagtta ggccaccact
     6721 tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg
     6781 ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata
     6841 aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga
     6901 cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag
     6961 ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg
     7021 agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac
     7081 ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca
     7141 acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg ttctttcctg
     7201 cgttatcccc tgattctgtg gataaccgta ttaccgcctt tgagtgagct gataccgctc
     7261 gccgcagccg aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa gagcgcccaa
     7321 tacgcaaacc gcctctcccc gcgcgttggc cgattcatta atgcagctgg cacgacaggt
     7381 ttcccgactg gaaagcgggc agtgagcgca acgcaattaa tgtgagttag ctcactcatt
     7441 aggcacccca ggctttacac tttatgcttc cggctcgtat gttgtgtgga attgtgagcg
     7501 gataacaatt tcacacagga aacagctatg accatgatta cgccaagctc tagctagagg
     7561 tcgaccaatt ctcatgtttg acagcttatc atcgcagatc cgggcaacgt tgttgccatt
     7621 gctgcaggcg cagaactggt aggtatggaa gatctataca ttgaatcaat attggcaatt
     7681 agccatatta gtcattggtt atatagcata aatcaatatt ggctattggc cattgcatac
     7741 gttgtatcta tatcataata tgtacattta tattggctca tgtccaatat gaccgccatg
     7801 ttgacattga ttattgacta gttattaata gtaatcaatt acggggtcat tagttcatag
     7861 cccatatatg gagttccgcg ttacataact tacggtaaat ggcccgcctg gctgaccgcc
     7921 caacgacccc cgcccattga cgtcaataat gacgtatgtt cccatagtaa cgccaatagg
     7981 gactttccat tgacgtcaat gggtggagta tttacggtaa actgcccact tggcagtaca
     8041 tcaagtgtat catatgccaa gtccgccccc tattgacgtc aatgacggta aatggcccgc
     8101 ctggcattat gcccagtaca tgaccttacg ggactttcct acttggcagt acatctacgt
     8161 attagtcatc gctattacca tggtgatgcg gttttggcag tacaccaatg ggcgtggata
     8221 gcggtttgac tcacggggat ttccaagtct ccaccccatt gacgtcaatg ggagtttgtt
     8281 ttggcaccaa aatcaacggg actttccaaa atgtcgtaat aaccccgccc cgttgacgca
     8341 aatgggcggt aggcgtgtac ggtgggaggt ctatataagc agagctcgtt tagtgaaccg
     8401 tcagatcctc actctcttcc gcatcgctgt ctgcgagggc cagctgttgg gctcgcggtt
     8461 gaggacaaac tcttcgcggt ctttccagta ctcttggatc ggaaacccgt cggcctccga
     8521 acggtactcc gccaccgagg gacctgagcg agtccgcatc gaccggatcg gaaaacct
//
