LOCUS       sequence-224557-        9152 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 9152)
  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 9152)
  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: TRAP-COMP-blac-flag-his
FEATURES             Location/Qualifiers
     source          1..9152
                     /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             2686..3471
                     /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             3478..3543
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             3550..3567
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    3603..3658
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(3657..3676)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(5894..5912)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5980..6084
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6085..6153,6154..6945)
                     /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      7116..7704
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7605..7624
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7858..7875
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    7992..8013
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(8028..8033,8034..8051,8052..8058)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8066..8082
                     /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     8071..8093
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     8090..8106
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     8090..8106
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     enhancer        8377..8756
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        8757..8960
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     8911..8931
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     8957..8981
                     /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 ggccgccaca
      421 atggcccatc tgggcaacgt gaagtacctc gtgatcgtgt tcctgatctt cttcgacctg
      481 ttcctcgtga acggccggga cgtgcagaac aacatcgtgg acgagatcaa gtaccgcgag
      541 gaagtgtgca acgacgaggt ggacctgtac ctgctgatgg actgcagcgg cagcatccgg
      601 cggcacaact gggtcaacca tgccgtgcct ctggccatga agctgatcca gcagctgaac
      661 ctgaacgaca acgccatcca cctgtacgcc agcgtgttca gcaacaacgc cagagagatc
      721 atcagactgc acagcgacgc cagcaagaac aaagagaagg ccctgatcat catcaagagc
      781 ctgctgagca ccaacctgcc ctacggcaag accaacctgg ccgatgctct gctgcaagtg
      841 cggaagcacc tgaatgaccg gatcaacaga gagaacgcca atcagctggt cgtgatcctg
      901 accgacggca tccccgacag catccaggac agcctgaaag agagccggaa gctgagcgac
      961 cggggcgtga agatcgccgt gtttggcatc ggacagggca tcaacgtggc cttcaaccgg
     1021 ttcctcgtgg gctgccaccc tagcgacggc aagtgcaatc tgtacgccga cagcgcctgg
     1081 gagaacgtga agaatgtgat cggccctttc atgaaggccg tgtgcgtgga agtggaaaag
     1141 accgccagct gcggcgtgtg ggatgagtgg tccccttgca gcgtgacctg cggcaagggc
     1201 accagaagcc ggaagagaga gatcctgcac gagggctgca ccagcgagct gcaggaacag
     1261 tgcgaagagg aacggtgcct gcccaagagg gaacccctgg atgtgcccga cgagcccgag
     1321 gacgatcagc ctagacccag aggcgacaac ttcgccgtgg aaaagcccaa cgagaacatc
     1381 atcgacaaca acccccagga acccagcccc aaccccgaag agggaaaggg cgagaacccc
     1441 aacggcttcg atctggacga gaaccctgag aaccccccca accctcccaa tccccctaac
     1501 cccccaaatc caccaaatcc tcccaaccct gacatccccg agcaggaacc taacatccct
     1561 gaggacagcg agaaagaggt gcccagcgac gtgcccaaga accccgagga tgaccgggaa
     1621 gagaacttcg acatccccaa gaagccagag aacaagcacg acaaccagaa caacctgcct
     1681 aacgacaagt ccgaccggta catcccctac agccccctga gccccaaggt gctggacaac
     1741 gagagaaagc agagcgaccc ccagagccag gacaacaacg gcaacagaca cgtgccaaac
     1801 agcgaggaca gagagacacg gccccacggc cggaacaacg agaatcgggc ctacaaccgg
     1861 aagcacaaca atgcccccaa gcaccctgag cgggaagaac acgaaaaacc tggcgcgccg
     1921 tcgacctcca tcacggccta taagagtgag ggggagtcag cggagttctc cttcccactc
     1981 aaccttggag aggaaagcct gcagggagag ttgagatgga aggcagagaa ggctccttct
     2041 tcccagtcct ggatcacctt ctccctaaag aaccaaaagg tgtctgtgca gaagtctact
     2101 agcaacccca agttccagct gtccgaaacg ctcccactca cccttcagat accccaggtc
     2161 tcccttcagt ttgctggttc tggcaacctg accctgactc tggacagagg gatactgtat
     2221 caggaagtga acctggtggt gatgaaagtg actcagcccg acagcaacac tttgacctgt
     2281 gaggtgatgg gacccacctc acccaagatg agactgatct tgaagcagga gaatcaggag
     2341 gccagggtct ccaggcagga gaaagtgatt caagtgcagg cccctgaagc aggggtgtgg
     2401 caatgtctac tgagtgaagg tgaagaggtc aagatggact ccaagatcca ggttttatcc
     2461 aaagggttga attccggcgg cggctccgga ggcggtaccg gtggcggcga cctagcccca
     2521 cagatgcttc gagaactcca ggagactaat gcggcgctgc aagacgtgag agagctcttg
     2581 cgacagcagg tcaaggagat caccttcctg aagaatacgg tgatggaatg tgacgcctgc
     2641 ggtctagaca gaaacctgcc tccactagca cctctgggcc cacacccaga aacgctggtg
     2701 aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg gttacatcga actggatctc
     2761 aacagcggta agatccttga gagttttcgc cccgaagaac gttttccaat gatgagcact
     2821 tttaaagttc tgctatgtgg cgcggtatta tcccgtattg acgccgggca agagcaactc
     2881 ggtcgccgca tacactattc tcagaatgac ttggttgagt actcaccagt cacagaaaag
     2941 catcttacgg atggcatgac agtaagagaa ttatgcagtg ctgccataac catgagtgat
     3001 aacactgcgg ccaacttact tctgacaacg atcggaggac cgaaggagct aaccgctttt
     3061 ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa
     3121 gccataccaa acgacgagcg tgacaccacg atgcctgtag caatggcaac aacgttgcgc
     3181 aaactattaa ctggcgaact acttactcta gcttcccggc aacaattaat agactggatg
     3241 gaggcggata aagttgcagg accacttctg cgctcggccc ttccggctgg ctggtttatt
     3301 gctgataaat ctggagccgg tgagcgtggg tctcgcggta tcattgcagc actggggcca
     3361 gatggtaagc cctcccgtat cgtagttatc tacacgacgg ggagtcaggc aactatggat
     3421 gaacgaaata gacagatcgc tgagataggt gcctcactga ttaagcattg gggccctgac
     3481 tacaaagacc atgacggtga ttataaagat catgacatcg attacaagga tgacgatgac
     3541 aagggccctc accaccacca tcaccactaa taataaggat cccccgacct cgacctctgg
     3601 ctaataaagg aaatttattt tcattgcaat agtgtgttgg aattttttgt gtctctcact
     3661 cggaaggaca tatgggaggg caaatcattt ggtcgagatc cctcggagat ctctagctag
     3721 aggatcgatc cccgccccgg acgaactaaa cctgactacg acatctctgc cccttcttcg
     3781 cggggcagtg catgtaatcc cttcagttgg ttggtacaac ttgccaactg ggccctgttc
     3841 cacatgtgac acgggggggg accaaacaca aaggggttct ctgactgtag ttgacatcct
     3901 tataaatgga tgtgcacatt tgccaacact gagtggcttt catcctggag cagactttgc
     3961 agtctgtgga ctgcaacaca acattgcctt tatgtgtaac tcttggctga agctcttaca
     4021 ccaatgctgg gggacatgta cctcccaggg gcccaggaag actacgggag gctacaccaa
     4081 cgtcaatcag aggggcctgt gtagctaccg ataagcggac cctcaagagg gcattagcaa
     4141 tagtgtttat aaggccccct tgttaaccct aaacgggtag catatgcttc ccgggtagta
     4201 gtatatacta tccagactaa ccctaattca atagcatatg ttacccaacg ggaagcatat
     4261 gctatcgaat tagggttagt aaaagggtcc taaggaacag cgatatctcc caccccatga
     4321 gctgtcacgg ttttatttac atggggtcag gattccacga gggtagtgaa ccattttagt
     4381 cacaagggca gtggctgaag atcaaggagc gggcagtgaa ctctcctgaa tcttcgcctg
     4441 cttcttcatt ctccttcgtt tagctaatag aataactgct gagttgtgaa cagtaaggtg
     4501 tatgtgaggt gctcgaaaac aaggtttcag gtgacgcccc cagaataaaa tttggacggg
     4561 gggttcagtg gtggcattgt gctatgacac caatataacc ctcacaaacc ccttgggcaa
     4621 taaatactag tgtaggaatg aaacattctg aatatcttta acaatagaaa tccatggggt
     4681 ggggacaagc cgtaaagact ggatgtccat ctcacacgaa tttatggcta tgggcaacac
     4741 ataatcctag tgcaatatga tactggggtt attaagatgt gtcccaggca gggaccaaga
     4801 caggtgaacc atgttgttac actctatttg taacaagggg aaagagagtg gacgccgaca
     4861 gcagcggact ccactggttg tctctaacac ccccgaaaat taaacggggc tccacgccaa
     4921 tggggcccat aaacaaagac aagtggccac tctttttttt gaaattgtgg agtgggggca
     4981 cgcgtcagcc cccacacgcc gccctgcggt tttggactgt aaaataaggg tgtaataact
     5041 tggctgattg taaccccgct aaccactgcg gtcaaaccac ttgcccacaa aaccactaat
     5101 ggcaccccgg ggaatacctg cataagtagg tgggcgggcc aagatagggg cgcgattgct
     5161 gcgatctgga ggacaaatta cacacacttg cgcctgagcg ccaagcacag ggttgttggt
     5221 cctcatattc acgaggtcgc tgagagcacg gtgggctaat gttgccatgg gtagcatata
     5281 ctacccaaat atctggatag catatgctat cctaatctat atctgggtag cataggctat
     5341 cctaatctat atctgggtag catatgctat cctaatctat atctgggtag tatatgctat
     5401 cctaatttat atctgggtag cataggctat cctaatctat atctgggtag catatgctat
     5461 cctaatctat atctgggtag tatatgctat cctaatctgt atccgggtag catatgctat
     5521 cctaatagag attagggtag tatatgctat cctaatttat atctgggtag catatactac
     5581 ccaaatatct ggatagcata tgctatccta atctatatct gggtagcata tgctatccta
     5641 atctatatct gggtagcata ggctatccta atctatatct gggtagcata tgctatccta
     5701 atctatatct gggtagtata tgctatccta atttatatct gggtagcata ggctatccta
     5761 atctatatct gggtagcata tgctatccta atctatatct gggtagtata tgctatccta
     5821 atctgtatcc gggtagcata tgctatcctc atgataagct gtcaaacatg agaattttct
     5881 tgaagacgaa agggcctcgt gatacgccta tttttatagg ttaatgtcat gataataatg
     5941 gtttcttaga cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta
     6001 tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt
     6061 caataatatt gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc ccttattccc
     6121 ttttttgcgg cattttgcct tcctgttttt gctcacccag aaacgctggt gaaagtaaaa
     6181 gatgctgaag atcagttggg tgcacgagtg ggttacatcg aactggatct caacagcggt
     6241 aagatccttg agagttttcg ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt
     6301 ctgctatgtg gcgcggtatt atcccgtgtt gacgccgggc aagagcaact cggtcgccgc
     6361 atacactatt ctcagaatga cttggttgag tactcaccag tcacagaaaa gcatcttacg
     6421 gatggcatga cagtaagaga attatgcagt gctgccataa ccatgagtga taacactgcg
     6481 gccaacttac ttctgacaac gatcggagga ccgaaggagc taaccgcttt tttgcacaac
     6541 atgggggatc atgtaactcg ccttgatcgt tgggaaccgg agctgaatga agccatacca
     6601 aacgacgagc gtgacaccac gatgcctgca gcaatggcaa caacgttgcg caaactatta
     6661 actggcgaac tacttactct agcttcccgg caacaattaa tagactggat ggaggcggat
     6721 aaagttgcag gaccacttct gcgctcggcc cttccggctg gctggtttat tgctgataaa
     6781 tctggagccg gtgagcgtgg gtctcgcggt atcattgcag cactggggcc agatggtaag
     6841 ccctcccgta tcgtagttat ctacacgacg gggagtcagg caactatgga tgaacgaaat
     6901 agacagatcg ctgagatagg tgcctcactg attaagcatt ggtaactgtc agaccaagtt
     6961 tactcatata tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg
     7021 aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga
     7081 gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta
     7141 atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa
     7201 gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact
     7261 gttcttctag tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca
     7321 tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt
     7381 accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg
     7441 ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag
     7501 cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta
     7561 agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat
     7621 ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg
     7681 tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc
     7741 ttttgctggc cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac
     7801 cgtattaccg cctttgagtg agctgatacc gctcgccgca gccgaacgac cgagcgcagc
     7861 gagtcagtga gcgaggaagc ggaagagcgc ccaatacgca aaccgcctct ccccgcgcgt
     7921 tggccgattc attaatgcag ctggcacgac aggtttcccg actggaaagc gggcagtgag
     7981 cgcaacgcaa ttaatgtgag ttagctcact cattaggcac cccaggcttt acactttatg
     8041 cttccggctc gtatgttgtg tggaattgtg agcggataac aatttcacac aggaaacagc
     8101 tatgaccatg attacgccaa gctctagcta gaggtcgacc aattctcatg tttgacagct
     8161 tatcatcgca gatccgggca acgttgttgc cattgctgca ggcgcagaac tggtaggtat
     8221 ggaagatcta tacattgaat caatattggc aattagccat attagtcatt ggttatatag
     8281 cataaatcaa tattggctat tggccattgc atacgttgta tctatatcat aatatgtaca
     8341 tttatattgg ctcatgtcca atatgaccgc catgttgaca ttgattattg actagttatt
     8401 aatagtaatc aattacgggg tcattagttc atagcccata tatggagttc cgcgttacat
     8461 aacttacggt aaatggcccg cctggctgac cgcccaacga cccccgccca ttgacgtcaa
     8521 taatgacgta tgttcccata gtaacgccaa tagggacttt ccattgacgt caatgggtgg
     8581 agtatttacg gtaaactgcc cacttggcag tacatcaagt gtatcatatg ccaagtccgc
     8641 cccctattga cgtcaatgac ggtaaatggc ccgcctggca ttatgcccag tacatgacct
     8701 tacgggactt tcctacttgg cagtacatct acgtattagt catcgctatt accatggtga
     8761 tgcggttttg gcagtacacc aatgggcgtg gatagcggtt tgactcacgg ggatttccaa
     8821 gtctccaccc cattgacgtc aatgggagtt tgttttggca ccaaaatcaa cgggactttc
     8881 caaaatgtcg taataacccc gccccgttga cgcaaatggg cggtaggcgt gtacggtggg
     8941 aggtctatat aagcagagct cgtttagtga accgtcagat cctcactctc ttccgcatcg
     9001 ctgtctgcga gggccagctg ttgggctcgc ggttgaggac aaactcttcg cggtctttcc
     9061 agtactcttg gatcggaaac ccgtcggcct ccgaacggta ctccgccacc gagggacctg
     9121 agcgagtccg catcgaccgg atcggaaaac ct
//
