LOCUS       sequence-208695-        8104 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 8104)
  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 8104)
  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_1352500-COMP-blac-flag-his
FEATURES             Location/Qualifiers
     source          1..8104
                     /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             1636..2421
                     /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             2428..2493
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             2500..2517
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    2555..2610
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(2609..2628)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(4846..4864)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4932..5036
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5037..5105,5106..5897)
                     /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      6068..6656
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6557..6576
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6810..6827
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    6944..6965
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6980..6985,6986..7003,7004..7010)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7018..7034
                     /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     7023..7045
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7042..7058
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7042..7058
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     enhancer        7329..7708
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        7709..7912
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     7863..7883
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     7909..7933
                     /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 ggccgcccag
      421 gacgtgatcg agctgaacga cgccaacttc gagagcctga cccagctgag caccggcaac
      481 accgccggca gctggttcat caagttctac gccccttggt gcagccactg caaggccatg
      541 agcaagacat gggcccagct ggccaccgag ctgaagggca agatcaacgt ggccaagatc
      601 gacgtgaccc tgaacagcaa gacccggaag cggttcaaga tcgagggctt ccccaccctg
      661 ctgtacttca agaacggcaa gatgtacgac tacaagaacc acgacagatc cctcgaggcc
      721 ttcaagaatt tcgtgctcga gacatacaag aacgccaagg ccagcgagcc tcccaagccc
      781 ctgaactaca tggatatcct gaaggacttc ctgaacgagg cctttcagaa catcgaccgg
      841 atctataagt acgccttccc cggcgcgccg tcgacctcca tcacggccta taagagtgag
      901 ggggagtcag cggagttctc cttcccactc aaccttggag aggaaagcct gcagggagag
      961 ttgagatgga aggcagagaa ggctccttct tcccagtcct ggatcacctt ctccctaaag
     1021 aaccaaaagg tgtctgtgca gaagtctact agcaacccca agttccagct gtccgaaacg
     1081 ctcccactca cccttcagat accccaggtc tcccttcagt ttgctggttc tggcaacctg
     1141 accctgactc tggacagagg gatactgtat caggaagtga acctggtggt gatgaaagtg
     1201 actcagcccg acagcaacac tttgacctgt gaggtgatgg gacccacctc acccaagatg
     1261 agactgatct tgaagcagga gaatcaggag gccagggtct ccaggcagga gaaagtgatt
     1321 caagtgcagg cccctgaagc aggggtgtgg caatgtctac tgagtgaagg tgaagaggtc
     1381 aagatggact ccaagatcca ggttttatcc aaagggttga attccggcgg cggctccgga
     1441 ggcggtaccg gtggcggcga cctagcccca cagatgcttc gagaactcca ggagactaat
     1501 gcggcgctgc aagacgtgag agagctcttg cgacagcagg tcaaggagat caccttcctg
     1561 aagaatacgg tgatggaatg tgacgcctgc ggtctagaca gaaacctgcc tccactagca
     1621 cctctgggcc cacacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt
     1681 gcacgagtgg gttacatcga actggatctc aacagcggta agatccttga gagttttcgc
     1741 cccgaagaac gttttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta
     1801 tcccgtattg acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac
     1861 ttggttgagt actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa
     1921 ttatgcagtg ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg
     1981 atcggaggac cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc
     2041 cttgatcgtt gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg
     2101 atgcctgtag caatggcaac aacgttgcgc aaactattaa ctggcgaact acttactcta
     2161 gcttcccggc aacaattaat agactggatg gaggcggata aagttgcagg accacttctg
     2221 cgctcggccc ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggg
     2281 tctcgcggta tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc
     2341 tacacgacgg ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt
     2401 gcctcactga ttaagcattg gggccctgac tacaaagacc atgacggtga ttataaagat
     2461 catgacatcg attacaagga tgacgatgac aagggccctc accaccacca tcaccactaa
     2521 ttaattaagg atcccccgac ctcgacctct ggctaataaa ggaaatttat tttcattgca
     2581 atagtgtgtt ggaatttttt gtgtctctca ctcggaagga catatgggag ggcaaatcat
     2641 ttggtcgaga tccctcggag atctctagct agaggatcga tccccgcccc ggacgaacta
     2701 aacctgacta cgacatctct gccccttctt cgcggggcag tgcatgtaat cccttcagtt
     2761 ggttggtaca acttgccaac tgggccctgt tccacatgtg acacgggggg ggaccaaaca
     2821 caaaggggtt ctctgactgt agttgacatc cttataaatg gatgtgcaca tttgccaaca
     2881 ctgagtggct ttcatcctgg agcagacttt gcagtctgtg gactgcaaca caacattgcc
     2941 tttatgtgta actcttggct gaagctctta caccaatgct gggggacatg tacctcccag
     3001 gggcccagga agactacggg aggctacacc aacgtcaatc agaggggcct gtgtagctac
     3061 cgataagcgg accctcaaga gggcattagc aatagtgttt ataaggcccc cttgttaacc
     3121 ctaaacgggt agcatatgct tcccgggtag tagtatatac tatccagact aaccctaatt
     3181 caatagcata tgttacccaa cgggaagcat atgctatcga attagggtta gtaaaagggt
     3241 cctaaggaac agcgatatct cccaccccat gagctgtcac ggttttattt acatggggtc
     3301 aggattccac gagggtagtg aaccatttta gtcacaaggg cagtggctga agatcaagga
     3361 gcgggcagtg aactctcctg aatcttcgcc tgcttcttca ttctccttcg tttagctaat
     3421 agaataactg ctgagttgtg aacagtaagg tgtatgtgag gtgctcgaaa acaaggtttc
     3481 aggtgacgcc cccagaataa aatttggacg gggggttcag tggtggcatt gtgctatgac
     3541 accaatataa ccctcacaaa ccccttgggc aataaatact agtgtaggaa tgaaacattc
     3601 tgaatatctt taacaataga aatccatggg gtggggacaa gccgtaaaga ctggatgtcc
     3661 atctcacacg aatttatggc tatgggcaac acataatcct agtgcaatat gatactgggg
     3721 ttattaagat gtgtcccagg cagggaccaa gacaggtgaa ccatgttgtt acactctatt
     3781 tgtaacaagg ggaaagagag tggacgccga cagcagcgga ctccactggt tgtctctaac
     3841 acccccgaaa attaaacggg gctccacgcc aatggggccc ataaacaaag acaagtggcc
     3901 actctttttt ttgaaattgt ggagtggggg cacgcgtcag cccccacacg ccgccctgcg
     3961 gttttggact gtaaaataag ggtgtaataa cttggctgat tgtaaccccg ctaaccactg
     4021 cggtcaaacc acttgcccac aaaaccacta atggcacccc ggggaatacc tgcataagta
     4081 ggtgggcggg ccaagatagg ggcgcgattg ctgcgatctg gaggacaaat tacacacact
     4141 tgcgcctgag cgccaagcac agggttgttg gtcctcatat tcacgaggtc gctgagagca
     4201 cggtgggcta atgttgccat gggtagcata tactacccaa atatctggat agcatatgct
     4261 atcctaatct atatctgggt agcataggct atcctaatct atatctgggt agcatatgct
     4321 atcctaatct atatctgggt agtatatgct atcctaattt atatctgggt agcataggct
     4381 atcctaatct atatctgggt agcatatgct atcctaatct atatctgggt agtatatgct
     4441 atcctaatct gtatccgggt agcatatgct atcctaatag agattagggt agtatatgct
     4501 atcctaattt atatctgggt agcatatact acccaaatat ctggatagca tatgctatcc
     4561 taatctatat ctgggtagca tatgctatcc taatctatat ctgggtagca taggctatcc
     4621 taatctatat ctgggtagca tatgctatcc taatctatat ctgggtagta tatgctatcc
     4681 taatttatat ctgggtagca taggctatcc taatctatat ctgggtagca tatgctatcc
     4741 taatctatat ctgggtagta tatgctatcc taatctgtat ccgggtagca tatgctatcc
     4801 tcatgataag ctgtcaaaca tgagaatttt cttgaagacg aaagggcctc gtgatacgcc
     4861 tatttttata ggttaatgtc atgataataa tggtttctta gacgtcaggt ggcacttttc
     4921 ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc
     4981 cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtatga
     5041 gtattcaaca tttccgtgtc gcccttattc ccttttttgc ggcattttgc cttcctgttt
     5101 ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga agatcagttg ggtgcacgag
     5161 tgggttacat cgaactggat ctcaacagcg gtaagatcct tgagagtttt cgccccgaag
     5221 aacgttttcc aatgatgagc acttttaaag ttctgctatg tggcgcggta ttatcccgtg
     5281 ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat gacttggttg
     5341 agtactcacc agtcacagaa aagcatctta cggatggcat gacagtaaga gaattatgca
     5401 gtgctgccat aaccatgagt gataacactg cggccaactt acttctgaca acgatcggag
     5461 gaccgaagga gctaaccgct tttttgcaca acatggggga tcatgtaact cgccttgatc
     5521 gttgggaacc ggagctgaat gaagccatac caaacgacga gcgtgacacc acgatgcctg
     5581 cagcaatggc aacaacgttg cgcaaactat taactggcga actacttact ctagcttccc
     5641 ggcaacaatt aatagactgg atggaggcgg ataaagttgc aggaccactt ctgcgctcgg
     5701 cccttccggc tggctggttt attgctgata aatctggagc cggtgagcgt gggtctcgcg
     5761 gtatcattgc agcactgggg ccagatggta agccctcccg tatcgtagtt atctacacga
     5821 cggggagtca ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac
     5881 tgattaagca ttggtaactg tcagaccaag tttactcata tatactttag attgatttaa
     5941 aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca
     6001 aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag
     6061 gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac
     6121 cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa
     6181 ctggcttcag cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttaggcc
     6241 accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag
     6301 tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac
     6361 cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc
     6421 gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc
     6481 ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca
     6541 cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc
     6601 tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg
     6661 ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgttct
     6721 ttcctgcgtt atcccctgat tctgtggata accgtattac cgcctttgag tgagctgata
     6781 ccgctcgccg cagccgaacg accgagcgca gcgagtcagt gagcgaggaa gcggaagagc
     6841 gcccaatacg caaaccgcct ctccccgcgc gttggccgat tcattaatgc agctggcacg
     6901 acaggtttcc cgactggaaa gcgggcagtg agcgcaacgc aattaatgtg agttagctca
     6961 ctcattaggc accccaggct ttacacttta tgcttccggc tcgtatgttg tgtggaattg
     7021 tgagcggata acaatttcac acaggaaaca gctatgacca tgattacgcc aagctctagc
     7081 tagaggtcga ccaattctca tgtttgacag cttatcatcg cagatccggg caacgttgtt
     7141 gccattgctg caggcgcaga actggtaggt atggaagatc tatacattga atcaatattg
     7201 gcaattagcc atattagtca ttggttatat agcataaatc aatattggct attggccatt
     7261 gcatacgttg tatctatatc ataatatgta catttatatt ggctcatgtc caatatgacc
     7321 gccatgttga cattgattat tgactagtta ttaatagtaa tcaattacgg ggtcattagt
     7381 tcatagccca tatatggagt tccgcgttac ataacttacg gtaaatggcc cgcctggctg
     7441 accgcccaac gacccccgcc cattgacgtc aataatgacg tatgttccca tagtaacgcc
     7501 aatagggact ttccattgac gtcaatgggt ggagtattta cggtaaactg cccacttggc
     7561 agtacatcaa gtgtatcata tgccaagtcc gccccctatt gacgtcaatg acggtaaatg
     7621 gcccgcctgg cattatgccc agtacatgac cttacgggac tttcctactt ggcagtacat
     7681 ctacgtatta gtcatcgcta ttaccatggt gatgcggttt tggcagtaca ccaatgggcg
     7741 tggatagcgg tttgactcac ggggatttcc aagtctccac cccattgacg tcaatgggag
     7801 tttgttttgg caccaaaatc aacgggactt tccaaaatgt cgtaataacc ccgccccgtt
     7861 gacgcaaatg ggcggtaggc gtgtacggtg ggaggtctat ataagcagag ctcgtttagt
     7921 gaaccgtcag atcctcactc tcttccgcat cgctgtctgc gagggccagc tgttgggctc
     7981 gcggttgagg acaaactctt cgcggtcttt ccagtactct tggatcggaa acccgtcggc
     8041 ctccgaacgg tactccgcca ccgagggacc tgagcgagtc cgcatcgacc ggatcggaaa
     8101 acct
//
