LOCUS       sequence-224273-        8576 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 8576)
  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 8576)
  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..8576
                     /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    3027..3082
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(3081..3100)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(5318..5336)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5404..5508
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5509..5577,5578..6369)
                     /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      6540..7128
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7029..7048
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7282..7299
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    7416..7437
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7452..7457,7458..7475,7476..7482)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7490..7506
                     /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     7495..7517
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7514..7530
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7514..7530
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     enhancer        7801..8180
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        8181..8384
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     8335..8355
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     8381..8405
                     /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 ctaataataa
     3001 ggatcccccg acctcgacct ctggctaata aaggaaattt attttcattg caatagtgtg
     3061 ttggaatttt ttgtgtctct cactcggaag gacatatggg agggcaaatc atttggtcga
     3121 gatccctcgg agatctctag ctagaggatc gatccccgcc ccggacgaac taaacctgac
     3181 tacgacatct ctgccccttc ttcgcggggc agtgcatgta atcccttcag ttggttggta
     3241 caacttgcca actgggccct gttccacatg tgacacgggg ggggaccaaa cacaaagggg
     3301 ttctctgact gtagttgaca tccttataaa tggatgtgca catttgccaa cactgagtgg
     3361 ctttcatcct ggagcagact ttgcagtctg tggactgcaa cacaacattg cctttatgtg
     3421 taactcttgg ctgaagctct tacaccaatg ctgggggaca tgtacctccc aggggcccag
     3481 gaagactacg ggaggctaca ccaacgtcaa tcagaggggc ctgtgtagct accgataagc
     3541 ggaccctcaa gagggcatta gcaatagtgt ttataaggcc cccttgttaa ccctaaacgg
     3601 gtagcatatg cttcccgggt agtagtatat actatccaga ctaaccctaa ttcaatagca
     3661 tatgttaccc aacgggaagc atatgctatc gaattagggt tagtaaaagg gtcctaagga
     3721 acagcgatat ctcccacccc atgagctgtc acggttttat ttacatgggg tcaggattcc
     3781 acgagggtag tgaaccattt tagtcacaag ggcagtggct gaagatcaag gagcgggcag
     3841 tgaactctcc tgaatcttcg cctgcttctt cattctcctt cgtttagcta atagaataac
     3901 tgctgagttg tgaacagtaa ggtgtatgtg aggtgctcga aaacaaggtt tcaggtgacg
     3961 cccccagaat aaaatttgga cggggggttc agtggtggca ttgtgctatg acaccaatat
     4021 aaccctcaca aaccccttgg gcaataaata ctagtgtagg aatgaaacat tctgaatatc
     4081 tttaacaata gaaatccatg gggtggggac aagccgtaaa gactggatgt ccatctcaca
     4141 cgaatttatg gctatgggca acacataatc ctagtgcaat atgatactgg ggttattaag
     4201 atgtgtccca ggcagggacc aagacaggtg aaccatgttg ttacactcta tttgtaacaa
     4261 ggggaaagag agtggacgcc gacagcagcg gactccactg gttgtctcta acacccccga
     4321 aaattaaacg gggctccacg ccaatggggc ccataaacaa agacaagtgg ccactctttt
     4381 ttttgaaatt gtggagtggg ggcacgcgtc agcccccaca cgccgccctg cggttttgga
     4441 ctgtaaaata agggtgtaat aacttggctg attgtaaccc cgctaaccac tgcggtcaaa
     4501 ccacttgccc acaaaaccac taatggcacc ccggggaata cctgcataag taggtgggcg
     4561 ggccaagata ggggcgcgat tgctgcgatc tggaggacaa attacacaca cttgcgcctg
     4621 agcgccaagc acagggttgt tggtcctcat attcacgagg tcgctgagag cacggtgggc
     4681 taatgttgcc atgggtagca tatactaccc aaatatctgg atagcatatg ctatcctaat
     4741 ctatatctgg gtagcatagg ctatcctaat ctatatctgg gtagcatatg ctatcctaat
     4801 ctatatctgg gtagtatatg ctatcctaat ttatatctgg gtagcatagg ctatcctaat
     4861 ctatatctgg gtagcatatg ctatcctaat ctatatctgg gtagtatatg ctatcctaat
     4921 ctgtatccgg gtagcatatg ctatcctaat agagattagg gtagtatatg ctatcctaat
     4981 ttatatctgg gtagcatata ctacccaaat atctggatag catatgctat cctaatctat
     5041 atctgggtag catatgctat cctaatctat atctgggtag cataggctat cctaatctat
     5101 atctgggtag catatgctat cctaatctat atctgggtag tatatgctat cctaatttat
     5161 atctgggtag cataggctat cctaatctat atctgggtag catatgctat cctaatctat
     5221 atctgggtag tatatgctat cctaatctgt atccgggtag catatgctat cctcatgata
     5281 agctgtcaaa catgagaatt ttcttgaaga cgaaagggcc tcgtgatacg cctattttta
     5341 taggttaatg tcatgataat aatggtttct tagacgtcag gtggcacttt tcggggaaat
     5401 gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg
     5461 agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtat gagtattcaa
     5521 catttccgtg tcgcccttat tccctttttt gcggcatttt gccttcctgt ttttgctcac
     5581 ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg agtgggttac
     5641 atcgaactgg atctcaacag cggtaagatc cttgagagtt ttcgccccga agaacgtttt
     5701 ccaatgatga gcacttttaa agttctgcta tgtggcgcgg tattatcccg tgttgacgcc
     5761 gggcaagagc aactcggtcg ccgcatacac tattctcaga atgacttggt tgagtactca
     5821 ccagtcacag aaaagcatct tacggatggc atgacagtaa gagaattatg cagtgctgcc
     5881 ataaccatga gtgataacac tgcggccaac ttacttctga caacgatcgg aggaccgaag
     5941 gagctaaccg cttttttgca caacatgggg gatcatgtaa ctcgccttga tcgttgggaa
     6001 ccggagctga atgaagccat accaaacgac gagcgtgaca ccacgatgcc tgcagcaatg
     6061 gcaacaacgt tgcgcaaact attaactggc gaactactta ctctagcttc ccggcaacaa
     6121 ttaatagact ggatggaggc ggataaagtt gcaggaccac ttctgcgctc ggcccttccg
     6181 gctggctggt ttattgctga taaatctgga gccggtgagc gtgggtctcg cggtatcatt
     6241 gcagcactgg ggccagatgg taagccctcc cgtatcgtag ttatctacac gacggggagt
     6301 caggcaacta tggatgaacg aaatagacag atcgctgaga taggtgcctc actgattaag
     6361 cattggtaac tgtcagacca agtttactca tatatacttt agattgattt aaaacttcat
     6421 ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac caaaatccct
     6481 taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct
     6541 tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca
     6601 gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc
     6661 agcagagcgc agataccaaa tactgttctt ctagtgtagc cgtagttagg ccaccacttc
     6721 aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct
     6781 gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag
     6841 gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc
     6901 tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg
     6961 agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag
     7021 cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt
     7081 gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac
     7141 gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt ctttcctgcg
     7201 ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga taccgctcgc
     7261 cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga gcgcccaata
     7321 cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctggca cgacaggttt
     7381 cccgactgga aagcgggcag tgagcgcaac gcaattaatg tgagttagct cactcattag
     7441 gcaccccagg ctttacactt tatgcttccg gctcgtatgt tgtgtggaat tgtgagcgga
     7501 taacaatttc acacaggaaa cagctatgac catgattacg ccaagctcta gctagaggtc
     7561 gaccaattct catgtttgac agcttatcat cgcagatccg ggcaacgttg ttgccattgc
     7621 tgcaggcgca gaactggtag gtatggaaga tctatacatt gaatcaatat tggcaattag
     7681 ccatattagt cattggttat atagcataaa tcaatattgg ctattggcca ttgcatacgt
     7741 tgtatctata tcataatatg tacatttata ttggctcatg tccaatatga ccgccatgtt
     7801 gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc
     7861 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
     7921 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
     7981 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
     8041 aagtgtatca tatgccaagt ccgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
     8101 ggcattatgc ccagtacatg accttacggg actttcctac ttggcagtac atctacgtat
     8161 tagtcatcgc tattaccatg gtgatgcggt tttggcagta caccaatggg cgtggatagc
     8221 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt
     8281 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaataa ccccgccccg ttgacgcaaa
     8341 tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctcgttta gtgaaccgtc
     8401 agatcctcac tctcttccgc atcgctgtct gcgagggcca gctgttgggc tcgcggttga
     8461 ggacaaactc ttcgcggtct ttccagtact cttggatcgg aaacccgtcg gcctccgaac
     8521 ggtactccgc caccgaggga cctgagcgag tccgcatcga ccggatcgga aaacct
//
