LOCUS       sequence-208825-        9172 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 9172)
  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 9172)
  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_1242000-COMP-blac-flag-his
FEATURES             Location/Qualifiers
     source          1..9172
                     /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             2704..3489
                     /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             3496..3561
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             3568..3585
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    3623..3678
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(3677..3696)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(5914..5932)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6000..6104
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6105..6173,6174..6965)
                     /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      7136..7724
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7625..7644
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7878..7895
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    8012..8033
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(8048..8053,8054..8071,8072..8078)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8086..8102
                     /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     8091..8113
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     8110..8126
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     8110..8126
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     enhancer        8397..8776
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        8777..8980
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     8931..8951
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     8977..9001
                     /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 ggccgccgat
      421 aatgtgtgca agggcgccga cgtgatcgtg aacgagttct ggtccaagct gaacaacatc
      481 cggcacgggg acgtgatcct gatcaacatc aagaactact actgccccgc ctgcaaccgg
      541 tacatgaaca tctggaacga cgtcgagaag aagatcctga cctacgagaa gaacgtggcc
      601 ctgttcgtgt tcgactgcag ctgctacctg ctggtgccct actgccggta cttcgacgtg
      661 ctgtacttcc ccaccttccg gctgctgttc cccgtgtacg acaagatcaa cgacaaagag
      721 tacaagtaca tcaaccccgc cagcatgatc gagggcgaga agtacaacgg cgacctgctg
      781 ctggcctaca gagaggtgga acggatcgac aacatctacg acttcaagat cctcatcgag
      841 aagtatctgt gcaagaacgt gaacttcaac tacaacgacc tgcggatctg catgaacagc
      901 gcctacatcc tgcccgagaa gaaaaacaag aagaagaacg agatctacca cttcctgttc
      961 ggctacaaga acgacaagaa caaagaagag atcaaggaca gcgtgaacgt gatgaacgac
     1021 gactttatcg agaacaacta cgattacgaa gaaaacaaag aaaacgtcga gcggtggtcc
     1081 aaccacatca acttcaacaa ggacaatatc aagcacgaca tcatcatcgg catcctgttc
     1141 accctgaaga agcacatcag cctgggccgg gacatccaga aagagtatat cgagcccttc
     1201 ctggtcatgc tgagcatcgt gtccaacatc tataacgacc tggaagaggg cctgtacgat
     1261 atcatcaaca agctcaacag cttcacctac cccattaagt acgacgagtg gatcaagtac
     1321 accgagaaca tcaattacat caacgagtac aagctggact acaacaacaa taacaacaac
     1381 gacgccaacc tgatcagctt caagctgtgc gagcagaaca gcgtgctgtg cagctactgg
     1441 ctgctgtatc acaagatcag cgtgtactgc ctgcagcacg acaagcacaa ctacctgtac
     1501 tacatcgagg ccatcaccaa ctacgccaag aattacctga actgccagaa ctgcatcgac
     1561 cactttctga acgcccagaa gttctgctac tacggctact gcaacatcca cagcgccgag
     1621 agcttcatca tctttctgtg gcggatccac aacgccgtga ccctgcggtc tatgtacgat
     1681 ctgctgatga tcgataacaa cgccgccacc cctagcaaca atttcaacaa gaagcagttc
     1741 ctgaaccagg acatcgtgtt ccctaccctg aagcagtgca agaattgccg gaacgccctg
     1801 ggcttcacca agatcaccaa tcacgccctg agcaccttca agaacggcgc catcaccgac
     1861 cagagcttcg acgccatcga cagcttcagc gtgaagaatg tgctgcacta cctgatccac
     1921 atctacagcg gcgcgccgtc gacctccatc acggcctata agagtgaggg ggagtcagcg
     1981 gagttctcct tcccactcaa ccttggagag gaaagcctgc agggagagtt gagatggaag
     2041 gcagagaagg ctccttcttc ccagtcctgg atcaccttct ccctaaagaa ccaaaaggtg
     2101 tctgtgcaga agtctactag caaccccaag ttccagctgt ccgaaacgct cccactcacc
     2161 cttcagatac cccaggtctc ccttcagttt gctggttctg gcaacctgac cctgactctg
     2221 gacagaggga tactgtatca ggaagtgaac ctggtggtga tgaaagtgac tcagcccgac
     2281 agcaacactt tgacctgtga ggtgatggga cccacctcac ccaagatgag actgatcttg
     2341 aagcaggaga atcaggaggc cagggtctcc aggcaggaga aagtgattca agtgcaggcc
     2401 cctgaagcag gggtgtggca atgtctactg agtgaaggtg aagaggtcaa gatggactcc
     2461 aagatccagg ttttatccaa agggttgaat tccggcggcg gctccggagg cggtaccggt
     2521 ggcggcgacc tagccccaca gatgcttcga gaactccagg agactaatgc ggcgctgcaa
     2581 gacgtgagag agctcttgcg acagcaggtc aaggagatca ccttcctgaa gaatacggtg
     2641 atggaatgtg acgcctgcgg tctagacaga aacctgcctc cactagcacc tctgggccca
     2701 cacccagaaa cgctggtgaa agtaaaagat gctgaagatc agttgggtgc acgagtgggt
     2761 tacatcgaac tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgt
     2821 tttccaatga tgagcacttt taaagttctg ctatgtggcg cggtattatc ccgtattgac
     2881 gccgggcaag agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtac
     2941 tcaccagtca cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct
     3001 gccataacca tgagtgataa cactgcggcc aacttacttc tgacaacgat cggaggaccg
     3061 aaggagctaa ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg
     3121 gaaccggagc tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgtagca
     3181 atggcaacaa cgttgcgcaa actattaact ggcgaactac ttactctagc ttcccggcaa
     3241 caattaatag actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt
     3301 ccggctggct ggtttattgc tgataaatct ggagccggtg agcgtgggtc tcgcggtatc
     3361 attgcagcac tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg
     3421 agtcaggcaa ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt
     3481 aagcattggg gccctgacta caaagaccat gacggtgatt ataaagatca tgacatcgat
     3541 tacaaggatg acgatgacaa gggccctcac caccaccatc accactaatt aattaaggat
     3601 cccccgacct cgacctctgg ctaataaagg aaatttattt tcattgcaat agtgtgttgg
     3661 aattttttgt gtctctcact cggaaggaca tatgggaggg caaatcattt ggtcgagatc
     3721 cctcggagat ctctagctag aggatcgatc cccgccccgg acgaactaaa cctgactacg
     3781 acatctctgc cccttcttcg cggggcagtg catgtaatcc cttcagttgg ttggtacaac
     3841 ttgccaactg ggccctgttc cacatgtgac acgggggggg accaaacaca aaggggttct
     3901 ctgactgtag ttgacatcct tataaatgga tgtgcacatt tgccaacact gagtggcttt
     3961 catcctggag cagactttgc agtctgtgga ctgcaacaca acattgcctt tatgtgtaac
     4021 tcttggctga agctcttaca ccaatgctgg gggacatgta cctcccaggg gcccaggaag
     4081 actacgggag gctacaccaa cgtcaatcag aggggcctgt gtagctaccg ataagcggac
     4141 cctcaagagg gcattagcaa tagtgtttat aaggccccct tgttaaccct aaacgggtag
     4201 catatgcttc ccgggtagta gtatatacta tccagactaa ccctaattca atagcatatg
     4261 ttacccaacg ggaagcatat gctatcgaat tagggttagt aaaagggtcc taaggaacag
     4321 cgatatctcc caccccatga gctgtcacgg ttttatttac atggggtcag gattccacga
     4381 gggtagtgaa ccattttagt cacaagggca gtggctgaag atcaaggagc gggcagtgaa
     4441 ctctcctgaa tcttcgcctg cttcttcatt ctccttcgtt tagctaatag aataactgct
     4501 gagttgtgaa cagtaaggtg tatgtgaggt gctcgaaaac aaggtttcag gtgacgcccc
     4561 cagaataaaa tttggacggg gggttcagtg gtggcattgt gctatgacac caatataacc
     4621 ctcacaaacc ccttgggcaa taaatactag tgtaggaatg aaacattctg aatatcttta
     4681 acaatagaaa tccatggggt ggggacaagc cgtaaagact ggatgtccat ctcacacgaa
     4741 tttatggcta tgggcaacac ataatcctag tgcaatatga tactggggtt attaagatgt
     4801 gtcccaggca gggaccaaga caggtgaacc atgttgttac actctatttg taacaagggg
     4861 aaagagagtg gacgccgaca gcagcggact ccactggttg tctctaacac ccccgaaaat
     4921 taaacggggc tccacgccaa tggggcccat aaacaaagac aagtggccac tctttttttt
     4981 gaaattgtgg agtgggggca cgcgtcagcc cccacacgcc gccctgcggt tttggactgt
     5041 aaaataaggg tgtaataact tggctgattg taaccccgct aaccactgcg gtcaaaccac
     5101 ttgcccacaa aaccactaat ggcaccccgg ggaatacctg cataagtagg tgggcgggcc
     5161 aagatagggg cgcgattgct gcgatctgga ggacaaatta cacacacttg cgcctgagcg
     5221 ccaagcacag ggttgttggt cctcatattc acgaggtcgc tgagagcacg gtgggctaat
     5281 gttgccatgg gtagcatata ctacccaaat atctggatag catatgctat cctaatctat
     5341 atctgggtag cataggctat cctaatctat atctgggtag catatgctat cctaatctat
     5401 atctgggtag tatatgctat cctaatttat atctgggtag cataggctat cctaatctat
     5461 atctgggtag catatgctat cctaatctat atctgggtag tatatgctat cctaatctgt
     5521 atccgggtag catatgctat cctaatagag attagggtag tatatgctat cctaatttat
     5581 atctgggtag catatactac ccaaatatct ggatagcata tgctatccta atctatatct
     5641 gggtagcata tgctatccta atctatatct gggtagcata ggctatccta atctatatct
     5701 gggtagcata tgctatccta atctatatct gggtagtata tgctatccta atttatatct
     5761 gggtagcata ggctatccta atctatatct gggtagcata tgctatccta atctatatct
     5821 gggtagtata tgctatccta atctgtatcc gggtagcata tgctatcctc atgataagct
     5881 gtcaaacatg agaattttct tgaagacgaa agggcctcgt gatacgccta tttttatagg
     5941 ttaatgtcat gataataatg gtttcttaga cgtcaggtgg cacttttcgg ggaaatgtgc
     6001 gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac
     6061 aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtatgagt attcaacatt
     6121 tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt gctcacccag
     6181 aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg ggttacatcg
     6241 aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa cgttttccaa
     6301 tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtgtt gacgccgggc
     6361 aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag tactcaccag
     6421 tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt gctgccataa
     6481 ccatgagtga taacactgcg gccaacttac ttctgacaac gatcggagga ccgaaggagc
     6541 taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt tgggaaccgg
     6601 agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgca gcaatggcaa
     6661 caacgttgcg caaactatta actggcgaac tacttactct agcttcccgg caacaattaa
     6721 tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc cttccggctg
     6781 gctggtttat tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt atcattgcag
     6841 cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg gggagtcagg
     6901 caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg attaagcatt
     6961 ggtaactgtc agaccaagtt tactcatata tactttagat tgatttaaaa cttcattttt
     7021 aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac
     7081 gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag
     7141 atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg
     7201 tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca
     7261 gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac cacttcaaga
     7321 actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca
     7381 gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc
     7441 agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca
     7501 ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa
     7561 aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc
     7621 cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc
     7681 gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg
     7741 cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt cctgcgttat
     7801 cccctgattc tgtggataac cgtattaccg cctttgagtg agctgatacc gctcgccgca
     7861 gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc ggaagagcgc ccaatacgca
     7921 aaccgcctct ccccgcgcgt tggccgattc attaatgcag ctggcacgac aggtttcccg
     7981 actggaaagc gggcagtgag cgcaacgcaa ttaatgtgag ttagctcact cattaggcac
     8041 cccaggcttt acactttatg cttccggctc gtatgttgtg tggaattgtg agcggataac
     8101 aatttcacac aggaaacagc tatgaccatg attacgccaa gctctagcta gaggtcgacc
     8161 aattctcatg tttgacagct tatcatcgca gatccgggca acgttgttgc cattgctgca
     8221 ggcgcagaac tggtaggtat ggaagatcta tacattgaat caatattggc aattagccat
     8281 attagtcatt ggttatatag cataaatcaa tattggctat tggccattgc atacgttgta
     8341 tctatatcat aatatgtaca tttatattgg ctcatgtcca atatgaccgc catgttgaca
     8401 ttgattattg actagttatt aatagtaatc aattacgggg tcattagttc atagcccata
     8461 tatggagttc cgcgttacat aacttacggt aaatggcccg cctggctgac cgcccaacga
     8521 cccccgccca ttgacgtcaa taatgacgta tgttcccata gtaacgccaa tagggacttt
     8581 ccattgacgt caatgggtgg agtatttacg gtaaactgcc cacttggcag tacatcaagt
     8641 gtatcatatg ccaagtccgc cccctattga cgtcaatgac ggtaaatggc ccgcctggca
     8701 ttatgcccag tacatgacct tacgggactt tcctacttgg cagtacatct acgtattagt
     8761 catcgctatt accatggtga tgcggttttg gcagtacacc aatgggcgtg gatagcggtt
     8821 tgactcacgg ggatttccaa gtctccaccc cattgacgtc aatgggagtt tgttttggca
     8881 ccaaaatcaa cgggactttc caaaatgtcg taataacccc gccccgttga cgcaaatggg
     8941 cggtaggcgt gtacggtggg aggtctatat aagcagagct cgtttagtga accgtcagat
     9001 cctcactctc ttccgcatcg ctgtctgcga gggccagctg ttgggctcgc ggttgaggac
     9061 aaactcttcg cggtctttcc agtactcttg gatcggaaac ccgtcggcct ccgaacggta
     9121 ctccgccacc gagggacctg agcgagtccg catcgaccgg atcggaaaac ct
//
