LOCUS       sequence-224282-        8093 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 8093)
  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 8093)
  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_0713700-COMP-blac-flag-his
FEATURES             Location/Qualifiers
     source          1..8093
                     /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             1627..2412
                     /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             2419..2484
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             2491..2508
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    2544..2599
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(2598..2617)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(4835..4853)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4921..5025
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5026..5094,5095..5886)
                     /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      6057..6645
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6546..6565
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6799..6816
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    6933..6954
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6969..6974,6975..6992,6993..6999)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7007..7023
                     /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     7012..7034
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7031..7047
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7031..7047
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     enhancer        7318..7697
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        7698..7901
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     7852..7872
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     7898..7922
                     /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 ggccgccgac
      421 atcatcatcg tggacagcaa caaggacatc cagggcttcg accccaacag cgtgctgaac
      481 ctggccgaga acgtgttcaa catcttcctg gacaccaaga ccaacaacgc cgatatcaag
      541 accgtgtaca gcagcatctt cagcaacagc aaagtgacca tcgacggcac ctgtagcagc
      601 atgaaggaac tgatcaaggg cagaagcctg aagtacctgg acgagctggg caccgaggaa
      661 tacgtgaaca gcgacgatgt gttcaacttc gccatcttcg tgtacaccgt gaacggccag
      721 cccgatatca tccagcaggt ctacaagaag caccccaact acatcagcta catcatcacc
      781 tgtaaagaga gcgagctgtt cgacaacatc gaggacgcca aggacaacaa gaatgtgttc
      841 cacaccaacc ccggcgcgcc gtcgacctcc atcacggcct ataagagtga gggggagtca
      901 gcggagttct ccttcccact caaccttgga gaggaaagcc tgcagggaga gttgagatgg
      961 aaggcagaga aggctccttc ttcccagtcc tggatcacct tctccctaaa gaaccaaaag
     1021 gtgtctgtgc agaagtctac tagcaacccc aagttccagc tgtccgaaac gctcccactc
     1081 acccttcaga taccccaggt ctcccttcag tttgctggtt ctggcaacct gaccctgact
     1141 ctggacagag ggatactgta tcaggaagtg aacctggtgg tgatgaaagt gactcagccc
     1201 gacagcaaca ctttgacctg tgaggtgatg ggacccacct cacccaagat gagactgatc
     1261 ttgaagcagg agaatcagga ggccagggtc tccaggcagg agaaagtgat tcaagtgcag
     1321 gcccctgaag caggggtgtg gcaatgtcta ctgagtgaag gtgaagaggt caagatggac
     1381 tccaagatcc aggttttatc caaagggttg aattccggcg gcggctccgg aggcggtacc
     1441 ggtggcggcg acctagcccc acagatgctt cgagaactcc aggagactaa tgcggcgctg
     1501 caagacgtga gagagctctt gcgacagcag gtcaaggaga tcaccttcct gaagaatacg
     1561 gtgatggaat gtgacgcctg cggtctagac agaaacctgc ctccactagc acctctgggc
     1621 ccacacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg
     1681 ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa
     1741 cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt
     1801 gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag
     1861 tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt
     1921 gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac gatcggagga
     1981 ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt
     2041 tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgta
     2101 gcaatggcaa caacgttgcg caaactatta actggcgaac tacttactct agcttcccgg
     2161 caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc
     2221 cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt
     2281 atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg
     2341 gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg
     2401 attaagcatt ggggccctga ctacaaagac catgacggtg attataaaga tcatgacatc
     2461 gattacaagg atgacgatga caagggccct caccaccacc atcaccacta ataataagga
     2521 tcccccgacc tcgacctctg gctaataaag gaaatttatt ttcattgcaa tagtgtgttg
     2581 gaattttttg tgtctctcac tcggaaggac atatgggagg gcaaatcatt tggtcgagat
     2641 ccctcggaga tctctagcta gaggatcgat ccccgccccg gacgaactaa acctgactac
     2701 gacatctctg ccccttcttc gcggggcagt gcatgtaatc ccttcagttg gttggtacaa
     2761 cttgccaact gggccctgtt ccacatgtga cacggggggg gaccaaacac aaaggggttc
     2821 tctgactgta gttgacatcc ttataaatgg atgtgcacat ttgccaacac tgagtggctt
     2881 tcatcctgga gcagactttg cagtctgtgg actgcaacac aacattgcct ttatgtgtaa
     2941 ctcttggctg aagctcttac accaatgctg ggggacatgt acctcccagg ggcccaggaa
     3001 gactacggga ggctacacca acgtcaatca gaggggcctg tgtagctacc gataagcgga
     3061 ccctcaagag ggcattagca atagtgttta taaggccccc ttgttaaccc taaacgggta
     3121 gcatatgctt cccgggtagt agtatatact atccagacta accctaattc aatagcatat
     3181 gttacccaac gggaagcata tgctatcgaa ttagggttag taaaagggtc ctaaggaaca
     3241 gcgatatctc ccaccccatg agctgtcacg gttttattta catggggtca ggattccacg
     3301 agggtagtga accattttag tcacaagggc agtggctgaa gatcaaggag cgggcagtga
     3361 actctcctga atcttcgcct gcttcttcat tctccttcgt ttagctaata gaataactgc
     3421 tgagttgtga acagtaaggt gtatgtgagg tgctcgaaaa caaggtttca ggtgacgccc
     3481 ccagaataaa atttggacgg ggggttcagt ggtggcattg tgctatgaca ccaatataac
     3541 cctcacaaac cccttgggca ataaatacta gtgtaggaat gaaacattct gaatatcttt
     3601 aacaatagaa atccatgggg tggggacaag ccgtaaagac tggatgtcca tctcacacga
     3661 atttatggct atgggcaaca cataatccta gtgcaatatg atactggggt tattaagatg
     3721 tgtcccaggc agggaccaag acaggtgaac catgttgtta cactctattt gtaacaaggg
     3781 gaaagagagt ggacgccgac agcagcggac tccactggtt gtctctaaca cccccgaaaa
     3841 ttaaacgggg ctccacgcca atggggccca taaacaaaga caagtggcca ctcttttttt
     3901 tgaaattgtg gagtgggggc acgcgtcagc ccccacacgc cgccctgcgg ttttggactg
     3961 taaaataagg gtgtaataac ttggctgatt gtaaccccgc taaccactgc ggtcaaacca
     4021 cttgcccaca aaaccactaa tggcaccccg gggaatacct gcataagtag gtgggcgggc
     4081 caagataggg gcgcgattgc tgcgatctgg aggacaaatt acacacactt gcgcctgagc
     4141 gccaagcaca gggttgttgg tcctcatatt cacgaggtcg ctgagagcac ggtgggctaa
     4201 tgttgccatg ggtagcatat actacccaaa tatctggata gcatatgcta tcctaatcta
     4261 tatctgggta gcataggcta tcctaatcta tatctgggta gcatatgcta tcctaatcta
     4321 tatctgggta gtatatgcta tcctaattta tatctgggta gcataggcta tcctaatcta
     4381 tatctgggta gcatatgcta tcctaatcta tatctgggta gtatatgcta tcctaatctg
     4441 tatccgggta gcatatgcta tcctaataga gattagggta gtatatgcta tcctaattta
     4501 tatctgggta gcatatacta cccaaatatc tggatagcat atgctatcct aatctatatc
     4561 tgggtagcat atgctatcct aatctatatc tgggtagcat aggctatcct aatctatatc
     4621 tgggtagcat atgctatcct aatctatatc tgggtagtat atgctatcct aatttatatc
     4681 tgggtagcat aggctatcct aatctatatc tgggtagcat atgctatcct aatctatatc
     4741 tgggtagtat atgctatcct aatctgtatc cgggtagcat atgctatcct catgataagc
     4801 tgtcaaacat gagaattttc ttgaagacga aagggcctcg tgatacgcct atttttatag
     4861 gttaatgtca tgataataat ggtttcttag acgtcaggtg gcacttttcg gggaaatgtg
     4921 cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga
     4981 caataaccct gataaatgct tcaataatat tgaaaaagga agagtatgag tattcaacat
     5041 ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt tgctcaccca
     5101 gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt gggttacatc
     5161 gaactggatc tcaacagcgg taagatcctt gagagttttc gccccgaaga acgttttcca
     5221 atgatgagca cttttaaagt tctgctatgt ggcgcggtat tatcccgtgt tgacgccggg
     5281 caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga gtactcacca
     5341 gtcacagaaa agcatcttac ggatggcatg acagtaagag aattatgcag tgctgccata
     5401 accatgagtg ataacactgc ggccaactta cttctgacaa cgatcggagg accgaaggag
     5461 ctaaccgctt ttttgcacaa catgggggat catgtaactc gccttgatcg ttgggaaccg
     5521 gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgc agcaatggca
     5581 acaacgttgc gcaaactatt aactggcgaa ctacttactc tagcttcccg gcaacaatta
     5641 atagactgga tggaggcgga taaagttgca ggaccacttc tgcgctcggc ccttccggct
     5701 ggctggttta ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca
     5761 gcactggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac ggggagtcag
     5821 gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact gattaagcat
     5881 tggtaactgt cagaccaagt ttactcatat atactttaga ttgatttaaa acttcatttt
     5941 taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa
     6001 cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga
     6061 gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg
     6121 gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc
     6181 agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca ccacttcaag
     6241 aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc
     6301 agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg
     6361 cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac
     6421 accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga
     6481 aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt
     6541 ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag
     6601 cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg
     6661 gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta
     6721 tcccctgatt ctgtggataa ccgtattacc gcctttgagt gagctgatac cgctcgccgc
     6781 agccgaacga ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg cccaatacgc
     6841 aaaccgcctc tccccgcgcg ttggccgatt cattaatgca gctggcacga caggtttccc
     6901 gactggaaag cgggcagtga gcgcaacgca attaatgtga gttagctcac tcattaggca
     6961 ccccaggctt tacactttat gcttccggct cgtatgttgt gtggaattgt gagcggataa
     7021 caatttcaca caggaaacag ctatgaccat gattacgcca agctctagct agaggtcgac
     7081 caattctcat gtttgacagc ttatcatcgc agatccgggc aacgttgttg ccattgctgc
     7141 aggcgcagaa ctggtaggta tggaagatct atacattgaa tcaatattgg caattagcca
     7201 tattagtcat tggttatata gcataaatca atattggcta ttggccattg catacgttgt
     7261 atctatatca taatatgtac atttatattg gctcatgtcc aatatgaccg ccatgttgac
     7321 attgattatt gactagttat taatagtaat caattacggg gtcattagtt catagcccat
     7381 atatggagtt ccgcgttaca taacttacgg taaatggccc gcctggctga ccgcccaacg
     7441 acccccgccc attgacgtca ataatgacgt atgttcccat agtaacgcca atagggactt
     7501 tccattgacg tcaatgggtg gagtatttac ggtaaactgc ccacttggca gtacatcaag
     7561 tgtatcatat gccaagtccg ccccctattg acgtcaatga cggtaaatgg cccgcctggc
     7621 attatgccca gtacatgacc ttacgggact ttcctacttg gcagtacatc tacgtattag
     7681 tcatcgctat taccatggtg atgcggtttt ggcagtacac caatgggcgt ggatagcggt
     7741 ttgactcacg gggatttcca agtctccacc ccattgacgt caatgggagt ttgttttggc
     7801 accaaaatca acgggacttt ccaaaatgtc gtaataaccc cgccccgttg acgcaaatgg
     7861 gcggtaggcg tgtacggtgg gaggtctata taagcagagc tcgtttagtg aaccgtcaga
     7921 tcctcactct cttccgcatc gctgtctgcg agggccagct gttgggctcg cggttgagga
     7981 caaactcttc gcggtctttc cagtactctt ggatcggaaa cccgtcggcc tccgaacggt
     8041 actccgccac cgagggacct gagcgagtcc gcatcgaccg gatcggaaaa cct
//
