LOCUS       sequence-224569-        9653 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 9653)
  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 9653)
  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_1106200-COMP-blac-flag-his
FEATURES             Location/Qualifiers
     source          1..9653
                     /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             3187..3972
                     /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             3979..4044
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             4051..4068
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    4104..4159
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(4158..4177)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(6395..6413)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6481..6585
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6586..6654,6655..7446)
                     /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      7617..8205
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8106..8125
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     8359..8376
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    8493..8514
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(8529..8534,8535..8552,8553..8559)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8567..8583
                     /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     8572..8594
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     8591..8607
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     8591..8607
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     enhancer        8878..9257
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        9258..9461
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     9412..9432
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     9458..9482
                     /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 ggccgccaac
      421 aaagaaatcg ccatcctgtg cagcaaagtg cacctgaaga agcacatcta ctaccccaga
      481 aagggcagcc acttccacat ctacagcgtg gtggacaaca acgtgaacct caagaacaag
      541 aacgagtaca acagcaagct gaaggaccgg atcaagatct acgaacaggt gcacaacaac
      601 aagtgcaaga agatctactt cctgcccctg atcttcctga agaacaacaa taaggacgtg
      661 tacatcgagg accacaagga caagaacatc gataacaata ataacaacaa gaaagataac
      721 aagaaggacg acatcaacaa caacgacaag aagaacaaaa tcaacgacgc ccctgcccag
      781 aatatcgcca acgataagga taagaactac gccaacaact tcgagtatat cgagaacaac
      841 ctgctgaagc tgaaaaacaa cctggaagtg ttcagcagct atatcaacaa ggccgacgtg
      901 atcatcgtgc ccctgaacta ccacgacatc ctgaacaagc acctgatcga gatcatccct
      961 ttcaacaaca tcatcaacag cctggactac atcttcctgt tcaacaagaa tgactataac
     1021 aaagccgaca tcaatattaa caacaacaac aacaataaca acaacatgaa caagtacaac
     1081 atctgccctc cctacaacaa tatctacaat atcatcttct tcatctacaa ctacaacgac
     1141 caggacaacc ctctgaacat tgccagctac acccctagct ttttcaccaa caaaaaggac
     1201 aacattatcc acaaagagaa gttccagctg gtccacgtga tccagaagtt tctgaacgac
     1261 cactacaacc tcaaggatca gaatgagcac aagaagtacc tgaacatcaa ctttctgttc
     1321 accaaggaca atatcctgga cattaacaag atcgacaagg atatcgaaga gaacaagatg
     1381 ctgtactacc ctaagaagaa tatcctcaac aaagagacaa actacgtcaa cacagccaac
     1441 aacgctaaca atgccaacaa gcaggaagat cagaacctga tcaaatacgt ccacaagaac
     1501 aatgacgcct tctttaagaa gtacaagccc gacaccctga agaatctggc caacgagaac
     1561 tactttatct ataacttcct gagcagctac aagctggaca tcctcaaaga gtacatctac
     1621 attgccaagg acatcaccat cccaaacaac aatgtgatca agcacatcca gaacattaac
     1681 atcaagaaga agatccacaa tatcgagctg ctgctcaaca atttcaagaa ctgcaagctc
     1741 aacaaaaaca tcaatagcca cgtgaacgtg caggaaatca agaaaaagca gtacagctac
     1801 ctgctgcaca tcatcctgag cgacctgctc gagaagtacg acagcaatct ggaaattctg
     1861 aagaactacc tgtaccaggt gttcaaagag aatatccaga agatcaagat taactccgat
     1921 atcctgttcc agtttaagaa gcagatcaac cacatcgacc agctgttcga ctactacaac
     1981 tccaagatct tcatcctccc cagcaacaag aactatcagg cccagcagaa gtcctactac
     2041 atctattact actataagat caagctgctg cagctcctga accaggccgc caatgagatc
     2101 atcaattact acatcagcaa gggcctgtac gtgaagaatt acgccttcaa cgagttcagc
     2161 ttctccaaga agtataccct gatcaattac atcatcacct acatctttaa cctgattaag
     2221 aacaaagtga atatcaactt caactacctg agccccaacg ccttcggctt cagctcctac
     2281 aaggacgata tcagcctgag ccctaagaaa gacatcatga tcaccaccag cgagatggac
     2341 caggtccaga aaatcaacat ccccagaagc gcctacacca agaccatgct gatgatggac
     2401 aaggacagaa acggcgcgcc gtcgacctcc atcacggcct ataagagtga gggggagtca
     2461 gcggagttct ccttcccact caaccttgga gaggaaagcc tgcagggaga gttgagatgg
     2521 aaggcagaga aggctccttc ttcccagtcc tggatcacct tctccctaaa gaaccaaaag
     2581 gtgtctgtgc agaagtctac tagcaacccc aagttccagc tgtccgaaac gctcccactc
     2641 acccttcaga taccccaggt ctcccttcag tttgctggtt ctggcaacct gaccctgact
     2701 ctggacagag ggatactgta tcaggaagtg aacctggtgg tgatgaaagt gactcagccc
     2761 gacagcaaca ctttgacctg tgaggtgatg ggacccacct cacccaagat gagactgatc
     2821 ttgaagcagg agaatcagga ggccagggtc tccaggcagg agaaagtgat tcaagtgcag
     2881 gcccctgaag caggggtgtg gcaatgtcta ctgagtgaag gtgaagaggt caagatggac
     2941 tccaagatcc aggttttatc caaagggttg aattccggcg gcggctccgg aggcggtacc
     3001 ggtggcggcg acctagcccc acagatgctt cgagaactcc aggagactaa tgcggcgctg
     3061 caagacgtga gagagctctt gcgacagcag gtcaaggaga tcaccttcct gaagaatacg
     3121 gtgatggaat gtgacgcctg cggtctagac agaaacctgc ctccactagc acctctgggc
     3181 ccacacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg
     3241 ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa
     3301 cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt
     3361 gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag
     3421 tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt
     3481 gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac gatcggagga
     3541 ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt
     3601 tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgta
     3661 gcaatggcaa caacgttgcg caaactatta actggcgaac tacttactct agcttcccgg
     3721 caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc
     3781 cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt
     3841 atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg
     3901 gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg
     3961 attaagcatt ggggccctga ctacaaagac catgacggtg attataaaga tcatgacatc
     4021 gattacaagg atgacgatga caagggccct caccaccacc atcaccacta ataataagga
     4081 tcccccgacc tcgacctctg gctaataaag gaaatttatt ttcattgcaa tagtgtgttg
     4141 gaattttttg tgtctctcac tcggaaggac atatgggagg gcaaatcatt tggtcgagat
     4201 ccctcggaga tctctagcta gaggatcgat ccccgccccg gacgaactaa acctgactac
     4261 gacatctctg ccccttcttc gcggggcagt gcatgtaatc ccttcagttg gttggtacaa
     4321 cttgccaact gggccctgtt ccacatgtga cacggggggg gaccaaacac aaaggggttc
     4381 tctgactgta gttgacatcc ttataaatgg atgtgcacat ttgccaacac tgagtggctt
     4441 tcatcctgga gcagactttg cagtctgtgg actgcaacac aacattgcct ttatgtgtaa
     4501 ctcttggctg aagctcttac accaatgctg ggggacatgt acctcccagg ggcccaggaa
     4561 gactacggga ggctacacca acgtcaatca gaggggcctg tgtagctacc gataagcgga
     4621 ccctcaagag ggcattagca atagtgttta taaggccccc ttgttaaccc taaacgggta
     4681 gcatatgctt cccgggtagt agtatatact atccagacta accctaattc aatagcatat
     4741 gttacccaac gggaagcata tgctatcgaa ttagggttag taaaagggtc ctaaggaaca
     4801 gcgatatctc ccaccccatg agctgtcacg gttttattta catggggtca ggattccacg
     4861 agggtagtga accattttag tcacaagggc agtggctgaa gatcaaggag cgggcagtga
     4921 actctcctga atcttcgcct gcttcttcat tctccttcgt ttagctaata gaataactgc
     4981 tgagttgtga acagtaaggt gtatgtgagg tgctcgaaaa caaggtttca ggtgacgccc
     5041 ccagaataaa atttggacgg ggggttcagt ggtggcattg tgctatgaca ccaatataac
     5101 cctcacaaac cccttgggca ataaatacta gtgtaggaat gaaacattct gaatatcttt
     5161 aacaatagaa atccatgggg tggggacaag ccgtaaagac tggatgtcca tctcacacga
     5221 atttatggct atgggcaaca cataatccta gtgcaatatg atactggggt tattaagatg
     5281 tgtcccaggc agggaccaag acaggtgaac catgttgtta cactctattt gtaacaaggg
     5341 gaaagagagt ggacgccgac agcagcggac tccactggtt gtctctaaca cccccgaaaa
     5401 ttaaacgggg ctccacgcca atggggccca taaacaaaga caagtggcca ctcttttttt
     5461 tgaaattgtg gagtgggggc acgcgtcagc ccccacacgc cgccctgcgg ttttggactg
     5521 taaaataagg gtgtaataac ttggctgatt gtaaccccgc taaccactgc ggtcaaacca
     5581 cttgcccaca aaaccactaa tggcaccccg gggaatacct gcataagtag gtgggcgggc
     5641 caagataggg gcgcgattgc tgcgatctgg aggacaaatt acacacactt gcgcctgagc
     5701 gccaagcaca gggttgttgg tcctcatatt cacgaggtcg ctgagagcac ggtgggctaa
     5761 tgttgccatg ggtagcatat actacccaaa tatctggata gcatatgcta tcctaatcta
     5821 tatctgggta gcataggcta tcctaatcta tatctgggta gcatatgcta tcctaatcta
     5881 tatctgggta gtatatgcta tcctaattta tatctgggta gcataggcta tcctaatcta
     5941 tatctgggta gcatatgcta tcctaatcta tatctgggta gtatatgcta tcctaatctg
     6001 tatccgggta gcatatgcta tcctaataga gattagggta gtatatgcta tcctaattta
     6061 tatctgggta gcatatacta cccaaatatc tggatagcat atgctatcct aatctatatc
     6121 tgggtagcat atgctatcct aatctatatc tgggtagcat aggctatcct aatctatatc
     6181 tgggtagcat atgctatcct aatctatatc tgggtagtat atgctatcct aatttatatc
     6241 tgggtagcat aggctatcct aatctatatc tgggtagcat atgctatcct aatctatatc
     6301 tgggtagtat atgctatcct aatctgtatc cgggtagcat atgctatcct catgataagc
     6361 tgtcaaacat gagaattttc ttgaagacga aagggcctcg tgatacgcct atttttatag
     6421 gttaatgtca tgataataat ggtttcttag acgtcaggtg gcacttttcg gggaaatgtg
     6481 cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga
     6541 caataaccct gataaatgct tcaataatat tgaaaaagga agagtatgag tattcaacat
     6601 ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt tgctcaccca
     6661 gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt gggttacatc
     6721 gaactggatc tcaacagcgg taagatcctt gagagttttc gccccgaaga acgttttcca
     6781 atgatgagca cttttaaagt tctgctatgt ggcgcggtat tatcccgtgt tgacgccggg
     6841 caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga gtactcacca
     6901 gtcacagaaa agcatcttac ggatggcatg acagtaagag aattatgcag tgctgccata
     6961 accatgagtg ataacactgc ggccaactta cttctgacaa cgatcggagg accgaaggag
     7021 ctaaccgctt ttttgcacaa catgggggat catgtaactc gccttgatcg ttgggaaccg
     7081 gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgc agcaatggca
     7141 acaacgttgc gcaaactatt aactggcgaa ctacttactc tagcttcccg gcaacaatta
     7201 atagactgga tggaggcgga taaagttgca ggaccacttc tgcgctcggc ccttccggct
     7261 ggctggttta ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca
     7321 gcactggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac ggggagtcag
     7381 gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact gattaagcat
     7441 tggtaactgt cagaccaagt ttactcatat atactttaga ttgatttaaa acttcatttt
     7501 taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa
     7561 cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga
     7621 gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg
     7681 gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc
     7741 agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca ccacttcaag
     7801 aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc
     7861 agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg
     7921 cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac
     7981 accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga
     8041 aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt
     8101 ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag
     8161 cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg
     8221 gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta
     8281 tcccctgatt ctgtggataa ccgtattacc gcctttgagt gagctgatac cgctcgccgc
     8341 agccgaacga ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg cccaatacgc
     8401 aaaccgcctc tccccgcgcg ttggccgatt cattaatgca gctggcacga caggtttccc
     8461 gactggaaag cgggcagtga gcgcaacgca attaatgtga gttagctcac tcattaggca
     8521 ccccaggctt tacactttat gcttccggct cgtatgttgt gtggaattgt gagcggataa
     8581 caatttcaca caggaaacag ctatgaccat gattacgcca agctctagct agaggtcgac
     8641 caattctcat gtttgacagc ttatcatcgc agatccgggc aacgttgttg ccattgctgc
     8701 aggcgcagaa ctggtaggta tggaagatct atacattgaa tcaatattgg caattagcca
     8761 tattagtcat tggttatata gcataaatca atattggcta ttggccattg catacgttgt
     8821 atctatatca taatatgtac atttatattg gctcatgtcc aatatgaccg ccatgttgac
     8881 attgattatt gactagttat taatagtaat caattacggg gtcattagtt catagcccat
     8941 atatggagtt ccgcgttaca taacttacgg taaatggccc gcctggctga ccgcccaacg
     9001 acccccgccc attgacgtca ataatgacgt atgttcccat agtaacgcca atagggactt
     9061 tccattgacg tcaatgggtg gagtatttac ggtaaactgc ccacttggca gtacatcaag
     9121 tgtatcatat gccaagtccg ccccctattg acgtcaatga cggtaaatgg cccgcctggc
     9181 attatgccca gtacatgacc ttacgggact ttcctacttg gcagtacatc tacgtattag
     9241 tcatcgctat taccatggtg atgcggtttt ggcagtacac caatgggcgt ggatagcggt
     9301 ttgactcacg gggatttcca agtctccacc ccattgacgt caatgggagt ttgttttggc
     9361 accaaaatca acgggacttt ccaaaatgtc gtaataaccc cgccccgttg acgcaaatgg
     9421 gcggtaggcg tgtacggtgg gaggtctata taagcagagc tcgtttagtg aaccgtcaga
     9481 tcctcactct cttccgcatc gctgtctgcg agggccagct gttgggctcg cggttgagga
     9541 caaactcttc gcggtctttc cagtactctt ggatcggaaa cccgtcggcc tccgaacggt
     9601 actccgccac cgagggacct gagcgagtcc gcatcgaccg gatcggaaaa cct
//
