LOCUS       sequence-208824-        9013 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 9013)
  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 9013)
  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: POFUT2-COMP-blac-flag-his
FEATURES             Location/Qualifiers
     source          1..9013
                     /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             2545..3330
                     /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             3337..3402
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             3409..3426
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    3464..3519
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(3518..3537)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(5755..5773)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5841..5945
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5946..6014,6015..6806)
                     /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      6977..7565
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7466..7485
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7719..7736
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    7853..7874
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7889..7894,7895..7912,7913..7919)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7927..7943
                     /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     7932..7954
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7951..7967
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7951..7967
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     enhancer        8238..8617
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        8618..8821
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     8772..8792
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     8818..8842
                     /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 ggccgccgtg
      421 atctgcgtga cccctcagaa gctgatctgc ctgaaagaag atgtgtacct gggcgatttc
      481 ttcttcttcc tgaaacggaa aaagtacatt atgtacgacg tgaacatcgg cgagggcttc
      541 aacctgcaga aagagatctt ctaccggctg agcctggtca tctacaacct gaacaagaag
      601 gacaagatca acatctacta cctggtgctg cctccctggt gctacgtgac ccactggaac
      661 atccggaagg gcaacaacct gagatgggag ttcttcttca acaccgacat catgaagaaa
      721 gtgatcccca tcatcgagta cgaggaatac gagaagctgt acggcaacta cgccgatatc
      781 atgatcaaca gcaagtacat cctggacaac tacaaagaga agtccttcct gatcctgccc
      841 ttcgaggaat gcaacatcaa cgtgaacagg ttcaagcagt tctgcaagaa gtgcgagcac
      901 aagtacaacg tgctgtactc cggctactgc accaccatca acaccaagca gagcgagtgc
      961 tacagctaca acatgatcag caactacttc atcacctcta tcctcgagaa cctcttcctg
     1021 tacaatatcg ccagcgtgct gatcaagcag tccaccaaca tcctggtgcc cttcgtgaac
     1081 gagctgtacc agagcaacct ggaagatatc ctgctgttca acaacaagct gctgagctac
     1141 ggcaacaact acatcagcaa catcctcaag accaaccact acatctccag ccacctgaga
     1201 tacaccgact ttaagtacat ctcccggtac aacgtcccac ccatccacat tgccctgctg
     1261 aagctgctgt acatcatgtt catcaacaat tgccggatca tctttatcgc ctccgacgag
     1321 aaggtggaaa tccagaaagt cattaacaag gacttccacc agtacaagaa gcacttctac
     1381 ttctacaaca accagaacaa cctccacgag ggcgagttct ctatcatcga gcagtggatc
     1441 tgcacccggt cctacatctt catcggcaac atcttcagcc ggttcaccat gaacatcaac
     1501 tgggagcggc acctgatcaa caagggccag atcaatcaga acatcgacct gtgcagctac
     1561 cacatcaacg acgacaacga ccaggacatc aagaactcct acaagaagat cgtccacatc
     1621 tttaaccaca aggccctgca gaagatcaag aacatctacg acaattacgc cgaccgggac
     1681 aagaagtaca tcaataccat ctgctacaac tttctgagcc acttccccaa caaccgggcc
     1741 atctacagaa aagagtacat caccaacacc ggcgcgccgt cgacctccat cacggcctat
     1801 aagagtgagg gggagtcagc ggagttctcc ttcccactca accttggaga ggaaagcctg
     1861 cagggagagt tgagatggaa ggcagagaag gctccttctt cccagtcctg gatcaccttc
     1921 tccctaaaga accaaaaggt gtctgtgcag aagtctacta gcaaccccaa gttccagctg
     1981 tccgaaacgc tcccactcac ccttcagata ccccaggtct cccttcagtt tgctggttct
     2041 ggcaacctga ccctgactct ggacagaggg atactgtatc aggaagtgaa cctggtggtg
     2101 atgaaagtga ctcagcccga cagcaacact ttgacctgtg aggtgatggg acccacctca
     2161 cccaagatga gactgatctt gaagcaggag aatcaggagg ccagggtctc caggcaggag
     2221 aaagtgattc aagtgcaggc ccctgaagca ggggtgtggc aatgtctact gagtgaaggt
     2281 gaagaggtca agatggactc caagatccag gttttatcca aagggttgaa ttccggcggc
     2341 ggctccggag gcggtaccgg tggcggcgac ctagccccac agatgcttcg agaactccag
     2401 gagactaatg cggcgctgca agacgtgaga gagctcttgc gacagcaggt caaggagatc
     2461 accttcctga agaatacggt gatggaatgt gacgcctgcg gtctagacag aaacctgcct
     2521 ccactagcac ctctgggccc acacccagaa acgctggtga aagtaaaaga tgctgaagat
     2581 cagttgggtg cacgagtggg ttacatcgaa ctggatctca acagcggtaa gatccttgag
     2641 agttttcgcc ccgaagaacg ttttccaatg atgagcactt ttaaagttct gctatgtggc
     2701 gcggtattat cccgtattga cgccgggcaa gagcaactcg gtcgccgcat acactattct
     2761 cagaatgact tggttgagta ctcaccagtc acagaaaagc atcttacgga tggcatgaca
     2821 gtaagagaat tatgcagtgc tgccataacc atgagtgata acactgcggc caacttactt
     2881 ctgacaacga tcggaggacc gaaggagcta accgcttttt tgcacaacat gggggatcat
     2941 gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt
     3001 gacaccacga tgcctgtagc aatggcaaca acgttgcgca aactattaac tggcgaacta
     3061 cttactctag cttcccggca acaattaata gactggatgg aggcggataa agttgcagga
     3121 ccacttctgc gctcggccct tccggctggc tggtttattg ctgataaatc tggagccggt
     3181 gagcgtgggt ctcgcggtat cattgcagca ctggggccag atggtaagcc ctcccgtatc
     3241 gtagttatct acacgacggg gagtcaggca actatggatg aacgaaatag acagatcgct
     3301 gagataggtg cctcactgat taagcattgg ggccctgact acaaagacca tgacggtgat
     3361 tataaagatc atgacatcga ttacaaggat gacgatgaca agggccctca ccaccaccat
     3421 caccactaat taattaagga tcccccgacc tcgacctctg gctaataaag gaaatttatt
     3481 ttcattgcaa tagtgtgttg gaattttttg tgtctctcac tcggaaggac atatgggagg
     3541 gcaaatcatt tggtcgagat ccctcggaga tctctagcta gaggatcgat ccccgccccg
     3601 gacgaactaa acctgactac gacatctctg ccccttcttc gcggggcagt gcatgtaatc
     3661 ccttcagttg gttggtacaa cttgccaact gggccctgtt ccacatgtga cacggggggg
     3721 gaccaaacac aaaggggttc tctgactgta gttgacatcc ttataaatgg atgtgcacat
     3781 ttgccaacac tgagtggctt tcatcctgga gcagactttg cagtctgtgg actgcaacac
     3841 aacattgcct ttatgtgtaa ctcttggctg aagctcttac accaatgctg ggggacatgt
     3901 acctcccagg ggcccaggaa gactacggga ggctacacca acgtcaatca gaggggcctg
     3961 tgtagctacc gataagcgga ccctcaagag ggcattagca atagtgttta taaggccccc
     4021 ttgttaaccc taaacgggta gcatatgctt cccgggtagt agtatatact atccagacta
     4081 accctaattc aatagcatat gttacccaac gggaagcata tgctatcgaa ttagggttag
     4141 taaaagggtc ctaaggaaca gcgatatctc ccaccccatg agctgtcacg gttttattta
     4201 catggggtca ggattccacg agggtagtga accattttag tcacaagggc agtggctgaa
     4261 gatcaaggag cgggcagtga actctcctga atcttcgcct gcttcttcat tctccttcgt
     4321 ttagctaata gaataactgc tgagttgtga acagtaaggt gtatgtgagg tgctcgaaaa
     4381 caaggtttca ggtgacgccc ccagaataaa atttggacgg ggggttcagt ggtggcattg
     4441 tgctatgaca ccaatataac cctcacaaac cccttgggca ataaatacta gtgtaggaat
     4501 gaaacattct gaatatcttt aacaatagaa atccatgggg tggggacaag ccgtaaagac
     4561 tggatgtcca tctcacacga atttatggct atgggcaaca cataatccta gtgcaatatg
     4621 atactggggt tattaagatg tgtcccaggc agggaccaag acaggtgaac catgttgtta
     4681 cactctattt gtaacaaggg gaaagagagt ggacgccgac agcagcggac tccactggtt
     4741 gtctctaaca cccccgaaaa ttaaacgggg ctccacgcca atggggccca taaacaaaga
     4801 caagtggcca ctcttttttt tgaaattgtg gagtgggggc acgcgtcagc ccccacacgc
     4861 cgccctgcgg ttttggactg taaaataagg gtgtaataac ttggctgatt gtaaccccgc
     4921 taaccactgc ggtcaaacca cttgcccaca aaaccactaa tggcaccccg gggaatacct
     4981 gcataagtag gtgggcgggc caagataggg gcgcgattgc tgcgatctgg aggacaaatt
     5041 acacacactt gcgcctgagc gccaagcaca gggttgttgg tcctcatatt cacgaggtcg
     5101 ctgagagcac ggtgggctaa tgttgccatg ggtagcatat actacccaaa tatctggata
     5161 gcatatgcta tcctaatcta tatctgggta gcataggcta tcctaatcta tatctgggta
     5221 gcatatgcta tcctaatcta tatctgggta gtatatgcta tcctaattta tatctgggta
     5281 gcataggcta tcctaatcta tatctgggta gcatatgcta tcctaatcta tatctgggta
     5341 gtatatgcta tcctaatctg tatccgggta gcatatgcta tcctaataga gattagggta
     5401 gtatatgcta tcctaattta tatctgggta gcatatacta cccaaatatc tggatagcat
     5461 atgctatcct aatctatatc tgggtagcat atgctatcct aatctatatc tgggtagcat
     5521 aggctatcct aatctatatc tgggtagcat atgctatcct aatctatatc tgggtagtat
     5581 atgctatcct aatttatatc tgggtagcat aggctatcct aatctatatc tgggtagcat
     5641 atgctatcct aatctatatc tgggtagtat atgctatcct aatctgtatc cgggtagcat
     5701 atgctatcct catgataagc tgtcaaacat gagaattttc ttgaagacga aagggcctcg
     5761 tgatacgcct atttttatag gttaatgtca tgataataat ggtttcttag acgtcaggtg
     5821 gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa
     5881 atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga
     5941 agagtatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc
     6001 ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg
     6061 gtgcacgagt gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc
     6121 gccccgaaga acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat
     6181 tatcccgtgt tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg
     6241 acttggttga gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag
     6301 aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa
     6361 cgatcggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc
     6421 gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca
     6481 cgatgcctgc agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc
     6541 tagcttcccg gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc
     6601 tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg
     6661 ggtctcgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta
     6721 tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag
     6781 gtgcctcact gattaagcat tggtaactgt cagaccaagt ttactcatat atactttaga
     6841 ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc
     6901 tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa
     6961 agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa
     7021 aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc
     7081 cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt
     7141 agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc
     7201 tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac
     7261 gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca
     7321 gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg
     7381 ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag
     7441 gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt
     7501 ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat
     7561 ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc
     7621 acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gcctttgagt
     7681 gagctgatac cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg agcgaggaag
     7741 cggaagagcg cccaatacgc aaaccgcctc tccccgcgcg ttggccgatt cattaatgca
     7801 gctggcacga caggtttccc gactggaaag cgggcagtga gcgcaacgca attaatgtga
     7861 gttagctcac tcattaggca ccccaggctt tacactttat gcttccggct cgtatgttgt
     7921 gtggaattgt gagcggataa caatttcaca caggaaacag ctatgaccat gattacgcca
     7981 agctctagct agaggtcgac caattctcat gtttgacagc ttatcatcgc agatccgggc
     8041 aacgttgttg ccattgctgc aggcgcagaa ctggtaggta tggaagatct atacattgaa
     8101 tcaatattgg caattagcca tattagtcat tggttatata gcataaatca atattggcta
     8161 ttggccattg catacgttgt atctatatca taatatgtac atttatattg gctcatgtcc
     8221 aatatgaccg ccatgttgac attgattatt gactagttat taatagtaat caattacggg
     8281 gtcattagtt catagcccat atatggagtt ccgcgttaca taacttacgg taaatggccc
     8341 gcctggctga ccgcccaacg acccccgccc attgacgtca ataatgacgt atgttcccat
     8401 agtaacgcca atagggactt tccattgacg tcaatgggtg gagtatttac ggtaaactgc
     8461 ccacttggca gtacatcaag tgtatcatat gccaagtccg ccccctattg acgtcaatga
     8521 cggtaaatgg cccgcctggc attatgccca gtacatgacc ttacgggact ttcctacttg
     8581 gcagtacatc tacgtattag tcatcgctat taccatggtg atgcggtttt ggcagtacac
     8641 caatgggcgt ggatagcggt ttgactcacg gggatttcca agtctccacc ccattgacgt
     8701 caatgggagt ttgttttggc accaaaatca acgggacttt ccaaaatgtc gtaataaccc
     8761 cgccccgttg acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata taagcagagc
     8821 tcgtttagtg aaccgtcaga tcctcactct cttccgcatc gctgtctgcg agggccagct
     8881 gttgggctcg cggttgagga caaactcttc gcggtctttc cagtactctt ggatcggaaa
     8941 cccgtcggcc tccgaacggt actccgccac cgagggacct gagcgagtcc gcatcgaccg
     9001 gatcggaaaa cct
//
