LOCUS       sequence-208822-        9424 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 9424)
  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 9424)
  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_0624400-COMP-blac-flag-his
FEATURES             Location/Qualifiers
     source          1..9424
                     /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             2956..3741
                     /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             3748..3813
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             3820..3837
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    3875..3930
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(3929..3948)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(6166..6184)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6252..6356
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6357..6425,6426..7217)
                     /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      7388..7976
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7877..7896
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     8130..8147
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    8264..8285
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(8300..8305,8306..8323,8324..8330)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8338..8354
                     /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     8343..8365
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     8362..8378
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     8362..8378
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     enhancer        8649..9028
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        9029..9232
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     9183..9203
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     9229..9253
                     /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 ggccgccgat
      421 gagagaaaga tcggcgagca gtacgagtac atcaacctgc tgagcgacaa agagtaccgg
      481 aacaagctgc tggccctgcg gaagcggacc ttcaacacca agaataccaa gatcaacaac
      541 aataataaca ataacaacat tgccaacagc aacggcaaca aagtgaacga catctacaac
      601 aagaaaaact acgtgttctg cttctacggc gcctgcggct acgacaagcg gaacgtgctg
      661 gaaaagatct tccacatcac cctggaaaag gaccagctga tcaacctggg ctacaaccag
      721 cggctgggca tttggttcgg caaggacaag gacgagaacc tgaacatgat cctggacatc
      781 gacctgctcg aaaacaaaga gttcatcgag aagaacaacg aggccatcta caattacgag
      841 aagctgatcg agacaatcct caagaccacc aacaccatca tcatccctct gctgtacgag
      901 gacatccagt gcgtgaccca cgtgctgaac aagaacaaca agatcaccga cgggaagaac
      961 aaggagtaca acaatctgaa gtacaagctg cctatcaaga tcgagctgtt cctgaacgag
     1021 ctgaaccgga agatcgagga agagaagaaa aagaacgaga acgactactc caacgggaag
     1081 gacatcctcg acaaaaaaaa gaacaatgtg tacttcatct tcctcaacaa cgagaaagag
     1141 aacaagcgca actacgccta caacaacaac ttcatccagg aaatcaagct gcggtacaac
     1201 aacctcaact tctacttcat gaacgaaaaa gagatcaacc tcgccctgct gaagtccaac
     1261 aatatcgccg agtgcatcat cctgatggac aacatcgagc acgccctgaa ggacatgagc
     1321 accttcaaga acttcgccga cttcaacaag gcctgcgtgt acaataccta cgtgatcgaa
     1381 gagagctaca tcaagaccct gaactacttc gacaagtact tctacaccta cgagaagaat
     1441 atcgccatgg gcaagatcat taacaagtac aacatcctga gcaacgaact gatcaagaac
     1501 agcctgctgc tgttcaatct gctgaccctg gaacagagcg gcaccaagtt caagcagatc
     1561 atcttcgaga agctcaagaa tcggttccag atgagcatcc ggcggcagat cctgaagcag
     1621 ctcctgctgc tcgagaaaaa gtatatcgag aagggcaagc acatcattct gaacaagtac
     1681 tatatcaaga atgacaaaga gctgattaac aacgagcaca accagctgca catcctgaag
     1741 aaaaccctga acagccagat caagcaggat atccagaccc tgatccacaa gcagttctac
     1801 aacgagcccc actttaacaa cgccatcaac cagttcatct accagctcga ggacaagctg
     1861 gaccacaccc tcaacaatta ctatgagctg aacaaagccc ctttcaagaa gatttttgaa
     1921 aagaaaaaaa actaccagat gaaccagaaa aagaagaaga agtatatctt ccccagcatc
     1981 aacatgaacc tggccctgac cagcctgatc cggaagtccg gcctgggcaa cctgcagagc
     2041 tacttcgtgt acgacctggg cttcctgacc ttcgtgttcg gcctgctgaa cgaccgggac
     2101 acccctgagg tgcagcagca gggcgcccac gtgccattct tcaagttcca gcccaagatt
     2161 aacctgaagc tgaacttcaa aggcgcgccg tcgacctcca tcacggccta taagagtgag
     2221 ggggagtcag cggagttctc cttcccactc aaccttggag aggaaagcct gcagggagag
     2281 ttgagatgga aggcagagaa ggctccttct tcccagtcct ggatcacctt ctccctaaag
     2341 aaccaaaagg tgtctgtgca gaagtctact agcaacccca agttccagct gtccgaaacg
     2401 ctcccactca cccttcagat accccaggtc tcccttcagt ttgctggttc tggcaacctg
     2461 accctgactc tggacagagg gatactgtat caggaagtga acctggtggt gatgaaagtg
     2521 actcagcccg acagcaacac tttgacctgt gaggtgatgg gacccacctc acccaagatg
     2581 agactgatct tgaagcagga gaatcaggag gccagggtct ccaggcagga gaaagtgatt
     2641 caagtgcagg cccctgaagc aggggtgtgg caatgtctac tgagtgaagg tgaagaggtc
     2701 aagatggact ccaagatcca ggttttatcc aaagggttga attccggcgg cggctccgga
     2761 ggcggtaccg gtggcggcga cctagcccca cagatgcttc gagaactcca ggagactaat
     2821 gcggcgctgc aagacgtgag agagctcttg cgacagcagg tcaaggagat caccttcctg
     2881 aagaatacgg tgatggaatg tgacgcctgc ggtctagaca gaaacctgcc tccactagca
     2941 cctctgggcc cacacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt
     3001 gcacgagtgg gttacatcga actggatctc aacagcggta agatccttga gagttttcgc
     3061 cccgaagaac gttttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta
     3121 tcccgtattg acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac
     3181 ttggttgagt actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa
     3241 ttatgcagtg ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg
     3301 atcggaggac cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc
     3361 cttgatcgtt gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg
     3421 atgcctgtag caatggcaac aacgttgcgc aaactattaa ctggcgaact acttactcta
     3481 gcttcccggc aacaattaat agactggatg gaggcggata aagttgcagg accacttctg
     3541 cgctcggccc ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggg
     3601 tctcgcggta tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc
     3661 tacacgacgg ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt
     3721 gcctcactga ttaagcattg gggccctgac tacaaagacc atgacggtga ttataaagat
     3781 catgacatcg attacaagga tgacgatgac aagggccctc accaccacca tcaccactaa
     3841 ttaattaagg atcccccgac ctcgacctct ggctaataaa ggaaatttat tttcattgca
     3901 atagtgtgtt ggaatttttt gtgtctctca ctcggaagga catatgggag ggcaaatcat
     3961 ttggtcgaga tccctcggag atctctagct agaggatcga tccccgcccc ggacgaacta
     4021 aacctgacta cgacatctct gccccttctt cgcggggcag tgcatgtaat cccttcagtt
     4081 ggttggtaca acttgccaac tgggccctgt tccacatgtg acacgggggg ggaccaaaca
     4141 caaaggggtt ctctgactgt agttgacatc cttataaatg gatgtgcaca tttgccaaca
     4201 ctgagtggct ttcatcctgg agcagacttt gcagtctgtg gactgcaaca caacattgcc
     4261 tttatgtgta actcttggct gaagctctta caccaatgct gggggacatg tacctcccag
     4321 gggcccagga agactacggg aggctacacc aacgtcaatc agaggggcct gtgtagctac
     4381 cgataagcgg accctcaaga gggcattagc aatagtgttt ataaggcccc cttgttaacc
     4441 ctaaacgggt agcatatgct tcccgggtag tagtatatac tatccagact aaccctaatt
     4501 caatagcata tgttacccaa cgggaagcat atgctatcga attagggtta gtaaaagggt
     4561 cctaaggaac agcgatatct cccaccccat gagctgtcac ggttttattt acatggggtc
     4621 aggattccac gagggtagtg aaccatttta gtcacaaggg cagtggctga agatcaagga
     4681 gcgggcagtg aactctcctg aatcttcgcc tgcttcttca ttctccttcg tttagctaat
     4741 agaataactg ctgagttgtg aacagtaagg tgtatgtgag gtgctcgaaa acaaggtttc
     4801 aggtgacgcc cccagaataa aatttggacg gggggttcag tggtggcatt gtgctatgac
     4861 accaatataa ccctcacaaa ccccttgggc aataaatact agtgtaggaa tgaaacattc
     4921 tgaatatctt taacaataga aatccatggg gtggggacaa gccgtaaaga ctggatgtcc
     4981 atctcacacg aatttatggc tatgggcaac acataatcct agtgcaatat gatactgggg
     5041 ttattaagat gtgtcccagg cagggaccaa gacaggtgaa ccatgttgtt acactctatt
     5101 tgtaacaagg ggaaagagag tggacgccga cagcagcgga ctccactggt tgtctctaac
     5161 acccccgaaa attaaacggg gctccacgcc aatggggccc ataaacaaag acaagtggcc
     5221 actctttttt ttgaaattgt ggagtggggg cacgcgtcag cccccacacg ccgccctgcg
     5281 gttttggact gtaaaataag ggtgtaataa cttggctgat tgtaaccccg ctaaccactg
     5341 cggtcaaacc acttgcccac aaaaccacta atggcacccc ggggaatacc tgcataagta
     5401 ggtgggcggg ccaagatagg ggcgcgattg ctgcgatctg gaggacaaat tacacacact
     5461 tgcgcctgag cgccaagcac agggttgttg gtcctcatat tcacgaggtc gctgagagca
     5521 cggtgggcta atgttgccat gggtagcata tactacccaa atatctggat agcatatgct
     5581 atcctaatct atatctgggt agcataggct atcctaatct atatctgggt agcatatgct
     5641 atcctaatct atatctgggt agtatatgct atcctaattt atatctgggt agcataggct
     5701 atcctaatct atatctgggt agcatatgct atcctaatct atatctgggt agtatatgct
     5761 atcctaatct gtatccgggt agcatatgct atcctaatag agattagggt agtatatgct
     5821 atcctaattt atatctgggt agcatatact acccaaatat ctggatagca tatgctatcc
     5881 taatctatat ctgggtagca tatgctatcc taatctatat ctgggtagca taggctatcc
     5941 taatctatat ctgggtagca tatgctatcc taatctatat ctgggtagta tatgctatcc
     6001 taatttatat ctgggtagca taggctatcc taatctatat ctgggtagca tatgctatcc
     6061 taatctatat ctgggtagta tatgctatcc taatctgtat ccgggtagca tatgctatcc
     6121 tcatgataag ctgtcaaaca tgagaatttt cttgaagacg aaagggcctc gtgatacgcc
     6181 tatttttata ggttaatgtc atgataataa tggtttctta gacgtcaggt ggcacttttc
     6241 ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc
     6301 cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtatga
     6361 gtattcaaca tttccgtgtc gcccttattc ccttttttgc ggcattttgc cttcctgttt
     6421 ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga agatcagttg ggtgcacgag
     6481 tgggttacat cgaactggat ctcaacagcg gtaagatcct tgagagtttt cgccccgaag
     6541 aacgttttcc aatgatgagc acttttaaag ttctgctatg tggcgcggta ttatcccgtg
     6601 ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat gacttggttg
     6661 agtactcacc agtcacagaa aagcatctta cggatggcat gacagtaaga gaattatgca
     6721 gtgctgccat aaccatgagt gataacactg cggccaactt acttctgaca acgatcggag
     6781 gaccgaagga gctaaccgct tttttgcaca acatggggga tcatgtaact cgccttgatc
     6841 gttgggaacc ggagctgaat gaagccatac caaacgacga gcgtgacacc acgatgcctg
     6901 cagcaatggc aacaacgttg cgcaaactat taactggcga actacttact ctagcttccc
     6961 ggcaacaatt aatagactgg atggaggcgg ataaagttgc aggaccactt ctgcgctcgg
     7021 cccttccggc tggctggttt attgctgata aatctggagc cggtgagcgt gggtctcgcg
     7081 gtatcattgc agcactgggg ccagatggta agccctcccg tatcgtagtt atctacacga
     7141 cggggagtca ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac
     7201 tgattaagca ttggtaactg tcagaccaag tttactcata tatactttag attgatttaa
     7261 aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca
     7321 aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag
     7381 gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac
     7441 cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa
     7501 ctggcttcag cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttaggcc
     7561 accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag
     7621 tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac
     7681 cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc
     7741 gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc
     7801 ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca
     7861 cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc
     7921 tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg
     7981 ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgttct
     8041 ttcctgcgtt atcccctgat tctgtggata accgtattac cgcctttgag tgagctgata
     8101 ccgctcgccg cagccgaacg accgagcgca gcgagtcagt gagcgaggaa gcggaagagc
     8161 gcccaatacg caaaccgcct ctccccgcgc gttggccgat tcattaatgc agctggcacg
     8221 acaggtttcc cgactggaaa gcgggcagtg agcgcaacgc aattaatgtg agttagctca
     8281 ctcattaggc accccaggct ttacacttta tgcttccggc tcgtatgttg tgtggaattg
     8341 tgagcggata acaatttcac acaggaaaca gctatgacca tgattacgcc aagctctagc
     8401 tagaggtcga ccaattctca tgtttgacag cttatcatcg cagatccggg caacgttgtt
     8461 gccattgctg caggcgcaga actggtaggt atggaagatc tatacattga atcaatattg
     8521 gcaattagcc atattagtca ttggttatat agcataaatc aatattggct attggccatt
     8581 gcatacgttg tatctatatc ataatatgta catttatatt ggctcatgtc caatatgacc
     8641 gccatgttga cattgattat tgactagtta ttaatagtaa tcaattacgg ggtcattagt
     8701 tcatagccca tatatggagt tccgcgttac ataacttacg gtaaatggcc cgcctggctg
     8761 accgcccaac gacccccgcc cattgacgtc aataatgacg tatgttccca tagtaacgcc
     8821 aatagggact ttccattgac gtcaatgggt ggagtattta cggtaaactg cccacttggc
     8881 agtacatcaa gtgtatcata tgccaagtcc gccccctatt gacgtcaatg acggtaaatg
     8941 gcccgcctgg cattatgccc agtacatgac cttacgggac tttcctactt ggcagtacat
     9001 ctacgtatta gtcatcgcta ttaccatggt gatgcggttt tggcagtaca ccaatgggcg
     9061 tggatagcgg tttgactcac ggggatttcc aagtctccac cccattgacg tcaatgggag
     9121 tttgttttgg caccaaaatc aacgggactt tccaaaatgt cgtaataacc ccgccccgtt
     9181 gacgcaaatg ggcggtaggc gtgtacggtg ggaggtctat ataagcagag ctcgtttagt
     9241 gaaccgtcag atcctcactc tcttccgcat cgctgtctgc gagggccagc tgttgggctc
     9301 gcggttgagg acaaactctt cgcggtcttt ccagtactct tggatcggaa acccgtcggc
     9361 ctccgaacgg tactccgcca ccgagggacc tgagcgagtc cgcatcgacc ggatcggaaa
     9421 acct
//
