LOCUS       sequence-208685-        9631 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 9631)
  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 9631)
  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_1105300-COMP-blac-flag-his
FEATURES             Location/Qualifiers
     source          1..9631
                     /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             3163..3948
                     /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             3955..4020
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             4027..4044
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    4082..4137
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(4136..4155)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(6373..6391)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6459..6563
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6564..6632,6633..7424)
                     /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      7595..8183
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8084..8103
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     8337..8354
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    8471..8492
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(8507..8512,8513..8530,8531..8537)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8545..8561
                     /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     8550..8572
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     8569..8585
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     8569..8585
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     enhancer        8856..9235
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        9236..9439
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     9390..9410
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     9436..9460
                     /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 ggccgcctac
      421 aagtacaccg tggaaggcaa agaagtgaaa gtcatctaca ccaacgacgc caacaagaac
      481 aagaagaaaa acgagctgaa caacttccag tacatcttcc tgaacgagaa cgaggaaaac
      541 tacaatatcg ccagcagcga cctgaacgac atcaacatcg aggaagagga catctacttc
      601 tacaagagct tcaccaagat catcaactac ttcaagtacc ggatccccgc caacatcaac
      661 ttcgccttcg agtacaacga cggcgacatc atcctgctgg acaaaaagaa aaagggcgtg
      721 tcctgcaaga tcaacaacct gttctacagc gagagcggcc tgaagtacct ggaccggaag
      781 aacgagctgt acgtgaagtc cgaggccttc gtgatcaagc acggcaacca ggaaaacaac
      841 gctaacaacg acaagaacga gggcatgaac caccggaaca acaataataa caataacaac
      901 tacaacaaca attataacaa caacaacaat tacaacaact actacaacaa cgccttcaac
      961 agcgccgtgc gcgaggacga gttcatcagc ggcaccgccc tgtgcgagaa cagaaacaag
     1021 aactacctgc ccaagaagat gctctgcttc cccagcagca acaaaaacaa tgagctgaat
     1081 atcgccttca tcaagaacga agagtacaaa gaactgatgg aactgtacat gaccggcatc
     1141 aagaaggaca acacctatct gctgaagaag atcgccaatt acttcaagag catcctgaac
     1201 ctggactacg acgtgaagaa tattgcctac tacatcttct gcaagaccct gcagtgcaac
     1261 ctgcagaacg tgttcgtgct gccctacgag tctgaggaag agtataagat ccagaacatc
     1321 atctggtcct ataacaacga gcagatcgcc ctgaacaatg ccttcatgta caacaatatc
     1381 ctggggaacg agaagatcgg caaccggaat atcatctggc ggcacgagaa taacaacatc
     1441 gtgaagcaca acattgccaa gtgcaacagc atctttttca acgagatgga atattacaac
     1501 tgctacaaga acctcgagaa cgtgtacgag atcctgctct acaaagagac aaacgtcctg
     1561 aacaagtaca tcaccaacaa ggccatcatt aagaacattg cctccgtgaa ccagtacacc
     1621 gccatcttct acgacgacaa cgactatgtg atcctgctga aggattacaa gtactacatc
     1681 agcttccagc tgaagaacat tatcgacctg gtgctgttcc aggacagcat caatgagatc
     1741 agcttctact acattaactt cgagagcaac aacatgctgc tgcggatcta caagtgcgcc
     1801 attaacaaca atgaagtgaa gtgcgccgac atcagctaca tcccttacga gatcaacgcc
     1861 aaagagaagt tcatctacct gagcaccaac cagaaaaagg atgtgtccag tgtgttcgtg
     1921 tccaccaaca ccaaagtgtg gaagatctat cacaccaagc aggaaaacga caacaacaac
     1981 aacaataaca ataacaacaa caacaataat aacaacaata acaataacaa caacaataat
     2041 aacaataaca acaacaataa caatgataac aatctgtaca gccgcaagct gatcgacagc
     2101 atccagaata ccaacgagga atacttcggc cacatcaatt gctacggcat cgaggccacc
     2161 tacgagagaa agctgttcag caagtatctg aacaacatcc ccgacgataa caagaacgtc
     2221 gtgggctgca gctacctgaa gtattacatc aatagcgcct ccaaggccct gctgatctcc
     2281 ttcatcgaga atgcccactt catccagcgg acctaccacg tcgtgaaaaa caagaatatc
     2341 attgtgtcct ccggcaccac aaagctgatc atcagagtga atgagctggg cgcgccgtcg
     2401 acctccatca cggcctataa gagtgagggg gagtcagcgg agttctcctt cccactcaac
     2461 cttggagagg aaagcctgca gggagagttg agatggaagg cagagaaggc tccttcttcc
     2521 cagtcctgga tcaccttctc cctaaagaac caaaaggtgt ctgtgcagaa gtctactagc
     2581 aaccccaagt tccagctgtc cgaaacgctc ccactcaccc ttcagatacc ccaggtctcc
     2641 cttcagtttg ctggttctgg caacctgacc ctgactctgg acagagggat actgtatcag
     2701 gaagtgaacc tggtggtgat gaaagtgact cagcccgaca gcaacacttt gacctgtgag
     2761 gtgatgggac ccacctcacc caagatgaga ctgatcttga agcaggagaa tcaggaggcc
     2821 agggtctcca ggcaggagaa agtgattcaa gtgcaggccc ctgaagcagg ggtgtggcaa
     2881 tgtctactga gtgaaggtga agaggtcaag atggactcca agatccaggt tttatccaaa
     2941 gggttgaatt ccggcggcgg ctccggaggc ggtaccggtg gcggcgacct agccccacag
     3001 atgcttcgag aactccagga gactaatgcg gcgctgcaag acgtgagaga gctcttgcga
     3061 cagcaggtca aggagatcac cttcctgaag aatacggtga tggaatgtga cgcctgcggt
     3121 ctagacagaa acctgcctcc actagcacct ctgggcccac acccagaaac gctggtgaaa
     3181 gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac
     3241 agcggtaaga tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt
     3301 aaagttctgc tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga gcaactcggt
     3361 cgccgcatac actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat
     3421 cttacggatg gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac
     3481 actgcggcca acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg
     3541 cacaacatgg gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc
     3601 ataccaaacg acgagcgtga caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa
     3661 ctattaactg gcgaactact tactctagct tcccggcaac aattaataga ctggatggag
     3721 gcggataaag ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct
     3781 gataaatctg gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat
     3841 ggtaagccct cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa
     3901 cgaaatagac agatcgctga gataggtgcc tcactgatta agcattgggg ccctgactac
     3961 aaagaccatg acggtgatta taaagatcat gacatcgatt acaaggatga cgatgacaag
     4021 ggccctcacc accaccatca ccactaatta attaaggatc ccccgacctc gacctctggc
     4081 taataaagga aatttatttt cattgcaata gtgtgttgga attttttgtg tctctcactc
     4141 ggaaggacat atgggagggc aaatcatttg gtcgagatcc ctcggagatc tctagctaga
     4201 ggatcgatcc ccgccccgga cgaactaaac ctgactacga catctctgcc ccttcttcgc
     4261 ggggcagtgc atgtaatccc ttcagttggt tggtacaact tgccaactgg gccctgttcc
     4321 acatgtgaca cgggggggga ccaaacacaa aggggttctc tgactgtagt tgacatcctt
     4381 ataaatggat gtgcacattt gccaacactg agtggctttc atcctggagc agactttgca
     4441 gtctgtggac tgcaacacaa cattgccttt atgtgtaact cttggctgaa gctcttacac
     4501 caatgctggg ggacatgtac ctcccagggg cccaggaaga ctacgggagg ctacaccaac
     4561 gtcaatcaga ggggcctgtg tagctaccga taagcggacc ctcaagaggg cattagcaat
     4621 agtgtttata aggccccctt gttaacccta aacgggtagc atatgcttcc cgggtagtag
     4681 tatatactat ccagactaac cctaattcaa tagcatatgt tacccaacgg gaagcatatg
     4741 ctatcgaatt agggttagta aaagggtcct aaggaacagc gatatctccc accccatgag
     4801 ctgtcacggt tttatttaca tggggtcagg attccacgag ggtagtgaac cattttagtc
     4861 acaagggcag tggctgaaga tcaaggagcg ggcagtgaac tctcctgaat cttcgcctgc
     4921 ttcttcattc tccttcgttt agctaataga ataactgctg agttgtgaac agtaaggtgt
     4981 atgtgaggtg ctcgaaaaca aggtttcagg tgacgccccc agaataaaat ttggacgggg
     5041 ggttcagtgg tggcattgtg ctatgacacc aatataaccc tcacaaaccc cttgggcaat
     5101 aaatactagt gtaggaatga aacattctga atatctttaa caatagaaat ccatggggtg
     5161 gggacaagcc gtaaagactg gatgtccatc tcacacgaat ttatggctat gggcaacaca
     5221 taatcctagt gcaatatgat actggggtta ttaagatgtg tcccaggcag ggaccaagac
     5281 aggtgaacca tgttgttaca ctctatttgt aacaagggga aagagagtgg acgccgacag
     5341 cagcggactc cactggttgt ctctaacacc cccgaaaatt aaacggggct ccacgccaat
     5401 ggggcccata aacaaagaca agtggccact cttttttttg aaattgtgga gtgggggcac
     5461 gcgtcagccc ccacacgccg ccctgcggtt ttggactgta aaataagggt gtaataactt
     5521 ggctgattgt aaccccgcta accactgcgg tcaaaccact tgcccacaaa accactaatg
     5581 gcaccccggg gaatacctgc ataagtaggt gggcgggcca agataggggc gcgattgctg
     5641 cgatctggag gacaaattac acacacttgc gcctgagcgc caagcacagg gttgttggtc
     5701 ctcatattca cgaggtcgct gagagcacgg tgggctaatg ttgccatggg tagcatatac
     5761 tacccaaata tctggatagc atatgctatc ctaatctata tctgggtagc ataggctatc
     5821 ctaatctata tctgggtagc atatgctatc ctaatctata tctgggtagt atatgctatc
     5881 ctaatttata tctgggtagc ataggctatc ctaatctata tctgggtagc atatgctatc
     5941 ctaatctata tctgggtagt atatgctatc ctaatctgta tccgggtagc atatgctatc
     6001 ctaatagaga ttagggtagt atatgctatc ctaatttata tctgggtagc atatactacc
     6061 caaatatctg gatagcatat gctatcctaa tctatatctg ggtagcatat gctatcctaa
     6121 tctatatctg ggtagcatag gctatcctaa tctatatctg ggtagcatat gctatcctaa
     6181 tctatatctg ggtagtatat gctatcctaa tttatatctg ggtagcatag gctatcctaa
     6241 tctatatctg ggtagcatat gctatcctaa tctatatctg ggtagtatat gctatcctaa
     6301 tctgtatccg ggtagcatat gctatcctca tgataagctg tcaaacatga gaattttctt
     6361 gaagacgaaa gggcctcgtg atacgcctat ttttataggt taatgtcatg ataataatgg
     6421 tttcttagac gtcaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat
     6481 ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc
     6541 aataatattg aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc cttattccct
     6601 tttttgcggc attttgcctt cctgtttttg ctcacccaga aacgctggtg aaagtaaaag
     6661 atgctgaaga tcagttgggt gcacgagtgg gttacatcga actggatctc aacagcggta
     6721 agatccttga gagttttcgc cccgaagaac gttttccaat gatgagcact tttaaagttc
     6781 tgctatgtgg cgcggtatta tcccgtgttg acgccgggca agagcaactc ggtcgccgca
     6841 tacactattc tcagaatgac ttggttgagt actcaccagt cacagaaaag catcttacgg
     6901 atggcatgac agtaagagaa ttatgcagtg ctgccataac catgagtgat aacactgcgg
     6961 ccaacttact tctgacaacg atcggaggac cgaaggagct aaccgctttt ttgcacaaca
     7021 tgggggatca tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa gccataccaa
     7081 acgacgagcg tgacaccacg atgcctgcag caatggcaac aacgttgcgc aaactattaa
     7141 ctggcgaact acttactcta gcttcccggc aacaattaat agactggatg gaggcggata
     7201 aagttgcagg accacttctg cgctcggccc ttccggctgg ctggtttatt gctgataaat
     7261 ctggagccgg tgagcgtggg tctcgcggta tcattgcagc actggggcca gatggtaagc
     7321 cctcccgtat cgtagttatc tacacgacgg ggagtcaggc aactatggat gaacgaaata
     7381 gacagatcgc tgagataggt gcctcactga ttaagcattg gtaactgtca gaccaagttt
     7441 actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga
     7501 agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag
     7561 cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa
     7621 tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag
     7681 agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg
     7741 ttcttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat
     7801 acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta
     7861 ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg
     7921 gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc
     7981 gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa
     8041 gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc
     8101 tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt
     8161 caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct
     8221 tttgctggcc ttttgctcac atgttctttc ctgcgttatc ccctgattct gtggataacc
     8281 gtattaccgc ctttgagtga gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg
     8341 agtcagtgag cgaggaagcg gaagagcgcc caatacgcaa accgcctctc cccgcgcgtt
     8401 ggccgattca ttaatgcagc tggcacgaca ggtttcccga ctggaaagcg ggcagtgagc
     8461 gcaacgcaat taatgtgagt tagctcactc attaggcacc ccaggcttta cactttatgc
     8521 ttccggctcg tatgttgtgt ggaattgtga gcggataaca atttcacaca ggaaacagct
     8581 atgaccatga ttacgccaag ctctagctag aggtcgacca attctcatgt ttgacagctt
     8641 atcatcgcag atccgggcaa cgttgttgcc attgctgcag gcgcagaact ggtaggtatg
     8701 gaagatctat acattgaatc aatattggca attagccata ttagtcattg gttatatagc
     8761 ataaatcaat attggctatt ggccattgca tacgttgtat ctatatcata atatgtacat
     8821 ttatattggc tcatgtccaa tatgaccgcc atgttgacat tgattattga ctagttatta
     8881 atagtaatca attacggggt cattagttca tagcccatat atggagttcc gcgttacata
     8941 acttacggta aatggcccgc ctggctgacc gcccaacgac ccccgcccat tgacgtcaat
     9001 aatgacgtat gttcccatag taacgccaat agggactttc cattgacgtc aatgggtgga
     9061 gtatttacgg taaactgccc acttggcagt acatcaagtg tatcatatgc caagtccgcc
     9121 ccctattgac gtcaatgacg gtaaatggcc cgcctggcat tatgcccagt acatgacctt
     9181 acgggacttt cctacttggc agtacatcta cgtattagtc atcgctatta ccatggtgat
     9241 gcggttttgg cagtacacca atgggcgtgg atagcggttt gactcacggg gatttccaag
     9301 tctccacccc attgacgtca atgggagttt gttttggcac caaaatcaac gggactttcc
     9361 aaaatgtcgt aataaccccg ccccgttgac gcaaatgggc ggtaggcgtg tacggtggga
     9421 ggtctatata agcagagctc gtttagtgaa ccgtcagatc ctcactctct tccgcatcgc
     9481 tgtctgcgag ggccagctgt tgggctcgcg gttgaggaca aactcttcgc ggtctttcca
     9541 gtactcttgg atcggaaacc cgtcggcctc cgaacggtac tccgccaccg agggacctga
     9601 gcgagtccgc atcgaccgga tcggaaaacc t
//
