LOCUS       sequence-224584-        8408 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 8408)
  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 8408)
  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..8408
                     /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    2859..2914
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(2913..2932)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(5150..5168)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5236..5340
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5341..5409,5410..6201)
                     /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      6372..6960
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6861..6880
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7114..7131
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    7248..7269
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7284..7289,7290..7307,7308..7314)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7322..7338
                     /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     7327..7349
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7346..7362
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7346..7362
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     enhancer        7633..8012
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        8013..8216
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     8167..8187
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     8213..8237
                     /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 cactaataat aaggatcccc cgacctcgac ctctggctaa taaaggaaat ttattttcat
     2881 tgcaatagtg tgttggaatt ttttgtgtct ctcactcgga aggacatatg ggagggcaaa
     2941 tcatttggtc gagatccctc ggagatctct agctagagga tcgatccccg ccccggacga
     3001 actaaacctg actacgacat ctctgcccct tcttcgcggg gcagtgcatg taatcccttc
     3061 agttggttgg tacaacttgc caactgggcc ctgttccaca tgtgacacgg ggggggacca
     3121 aacacaaagg ggttctctga ctgtagttga catccttata aatggatgtg cacatttgcc
     3181 aacactgagt ggctttcatc ctggagcaga ctttgcagtc tgtggactgc aacacaacat
     3241 tgcctttatg tgtaactctt ggctgaagct cttacaccaa tgctggggga catgtacctc
     3301 ccaggggccc aggaagacta cgggaggcta caccaacgtc aatcagaggg gcctgtgtag
     3361 ctaccgataa gcggaccctc aagagggcat tagcaatagt gtttataagg cccccttgtt
     3421 aaccctaaac gggtagcata tgcttcccgg gtagtagtat atactatcca gactaaccct
     3481 aattcaatag catatgttac ccaacgggaa gcatatgcta tcgaattagg gttagtaaaa
     3541 gggtcctaag gaacagcgat atctcccacc ccatgagctg tcacggtttt atttacatgg
     3601 ggtcaggatt ccacgagggt agtgaaccat tttagtcaca agggcagtgg ctgaagatca
     3661 aggagcgggc agtgaactct cctgaatctt cgcctgcttc ttcattctcc ttcgtttagc
     3721 taatagaata actgctgagt tgtgaacagt aaggtgtatg tgaggtgctc gaaaacaagg
     3781 tttcaggtga cgcccccaga ataaaatttg gacggggggt tcagtggtgg cattgtgcta
     3841 tgacaccaat ataaccctca caaacccctt gggcaataaa tactagtgta ggaatgaaac
     3901 attctgaata tctttaacaa tagaaatcca tggggtgggg acaagccgta aagactggat
     3961 gtccatctca cacgaattta tggctatggg caacacataa tcctagtgca atatgatact
     4021 ggggttatta agatgtgtcc caggcaggga ccaagacagg tgaaccatgt tgttacactc
     4081 tatttgtaac aaggggaaag agagtggacg ccgacagcag cggactccac tggttgtctc
     4141 taacaccccc gaaaattaaa cggggctcca cgccaatggg gcccataaac aaagacaagt
     4201 ggccactctt ttttttgaaa ttgtggagtg ggggcacgcg tcagccccca cacgccgccc
     4261 tgcggttttg gactgtaaaa taagggtgta ataacttggc tgattgtaac cccgctaacc
     4321 actgcggtca aaccacttgc ccacaaaacc actaatggca ccccggggaa tacctgcata
     4381 agtaggtggg cgggccaaga taggggcgcg attgctgcga tctggaggac aaattacaca
     4441 cacttgcgcc tgagcgccaa gcacagggtt gttggtcctc atattcacga ggtcgctgag
     4501 agcacggtgg gctaatgttg ccatgggtag catatactac ccaaatatct ggatagcata
     4561 tgctatccta atctatatct gggtagcata ggctatccta atctatatct gggtagcata
     4621 tgctatccta atctatatct gggtagtata tgctatccta atttatatct gggtagcata
     4681 ggctatccta atctatatct gggtagcata tgctatccta atctatatct gggtagtata
     4741 tgctatccta atctgtatcc gggtagcata tgctatccta atagagatta gggtagtata
     4801 tgctatccta atttatatct gggtagcata tactacccaa atatctggat agcatatgct
     4861 atcctaatct atatctgggt agcatatgct atcctaatct atatctgggt agcataggct
     4921 atcctaatct atatctgggt agcatatgct atcctaatct atatctgggt agtatatgct
     4981 atcctaattt atatctgggt agcataggct atcctaatct atatctgggt agcatatgct
     5041 atcctaatct atatctgggt agtatatgct atcctaatct gtatccgggt agcatatgct
     5101 atcctcatga taagctgtca aacatgagaa ttttcttgaa gacgaaaggg cctcgtgata
     5161 cgcctatttt tataggttaa tgtcatgata ataatggttt cttagacgtc aggtggcact
     5221 tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg
     5281 tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt
     5341 atgagtattc aacatttccg tgtcgccctt attccctttt ttgcggcatt ttgccttcct
     5401 gtttttgctc acccagaaac gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca
     5461 cgagtgggtt acatcgaact ggatctcaac agcggtaaga tccttgagag ttttcgcccc
     5521 gaagaacgtt ttccaatgat gagcactttt aaagttctgc tatgtggcgc ggtattatcc
     5581 cgtgttgacg ccgggcaaga gcaactcggt cgccgcatac actattctca gaatgacttg
     5641 gttgagtact caccagtcac agaaaagcat cttacggatg gcatgacagt aagagaatta
     5701 tgcagtgctg ccataaccat gagtgataac actgcggcca acttacttct gacaacgatc
     5761 ggaggaccga aggagctaac cgcttttttg cacaacatgg gggatcatgt aactcgcctt
     5821 gatcgttggg aaccggagct gaatgaagcc ataccaaacg acgagcgtga caccacgatg
     5881 cctgcagcaa tggcaacaac gttgcgcaaa ctattaactg gcgaactact tactctagct
     5941 tcccggcaac aattaataga ctggatggag gcggataaag ttgcaggacc acttctgcgc
     6001 tcggcccttc cggctggctg gtttattgct gataaatctg gagccggtga gcgtgggtct
     6061 cgcggtatca ttgcagcact ggggccagat ggtaagccct cccgtatcgt agttatctac
     6121 acgacgggga gtcaggcaac tatggatgaa cgaaatagac agatcgctga gataggtgcc
     6181 tcactgatta agcattggta actgtcagac caagtttact catatatact ttagattgat
     6241 ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga taatctcatg
     6301 accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc
     6361 aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa
     6421 ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag
     6481 gtaactggct tcagcagagc gcagatacca aatactgttc ttctagtgta gccgtagtta
     6541 ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta
     6601 ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag
     6661 ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg
     6721 gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg
     6781 cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag
     6841 cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc
     6901 cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa
     6961 aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg
     7021 ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgcctt tgagtgagct
     7081 gataccgctc gccgcagccg aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa
     7141 gagcgcccaa tacgcaaacc gcctctcccc gcgcgttggc cgattcatta atgcagctgg
     7201 cacgacaggt ttcccgactg gaaagcgggc agtgagcgca acgcaattaa tgtgagttag
     7261 ctcactcatt aggcacccca ggctttacac tttatgcttc cggctcgtat gttgtgtgga
     7321 attgtgagcg gataacaatt tcacacagga aacagctatg accatgatta cgccaagctc
     7381 tagctagagg tcgaccaatt ctcatgtttg acagcttatc atcgcagatc cgggcaacgt
     7441 tgttgccatt gctgcaggcg cagaactggt aggtatggaa gatctataca ttgaatcaat
     7501 attggcaatt agccatatta gtcattggtt atatagcata aatcaatatt ggctattggc
     7561 cattgcatac gttgtatcta tatcataata tgtacattta tattggctca tgtccaatat
     7621 gaccgccatg ttgacattga ttattgacta gttattaata gtaatcaatt acggggtcat
     7681 tagttcatag cccatatatg gagttccgcg ttacataact tacggtaaat ggcccgcctg
     7741 gctgaccgcc caacgacccc cgcccattga cgtcaataat gacgtatgtt cccatagtaa
     7801 cgccaatagg gactttccat tgacgtcaat gggtggagta tttacggtaa actgcccact
     7861 tggcagtaca tcaagtgtat catatgccaa gtccgccccc tattgacgtc aatgacggta
     7921 aatggcccgc ctggcattat gcccagtaca tgaccttacg ggactttcct acttggcagt
     7981 acatctacgt attagtcatc gctattacca tggtgatgcg gttttggcag tacaccaatg
     8041 ggcgtggata gcggtttgac tcacggggat ttccaagtct ccaccccatt gacgtcaatg
     8101 ggagtttgtt ttggcaccaa aatcaacggg actttccaaa atgtcgtaat aaccccgccc
     8161 cgttgacgca aatgggcggt aggcgtgtac ggtgggaggt ctatataagc agagctcgtt
     8221 tagtgaaccg tcagatcctc actctcttcc gcatcgctgt ctgcgagggc cagctgttgg
     8281 gctcgcggtt gaggacaaac tcttcgcggt ctttccagta ctcttggatc ggaaacccgt
     8341 cggcctccga acggtactcc gccaccgagg gacctgagcg agtccgcatc gaccggatcg
     8401 gaaaacct
//
