LOCUS       sequence-208821-        8791 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 8791)
  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 8791)
  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_0107300-COMP-blac-flag-his
FEATURES             Location/Qualifiers
     source          1..8791
                     /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             2323..3108
                     /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             3115..3180
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             3187..3204
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    3242..3297
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(3296..3315)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(5533..5551)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5619..5723
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5724..5792,5793..6584)
                     /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      6755..7343
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7244..7263
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7497..7514
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    7631..7652
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7667..7672,7673..7690,7691..7697)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7705..7721
                     /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     7710..7732
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7729..7745
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7729..7745
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     enhancer        8016..8395
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        8396..8599
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     8550..8570
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     8596..8620
                     /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 ggccgccatc
      421 aaccctgcca ccctgatcca gaaccacctg tacatgtatc agaccaacaa cgccatcaat
      481 gaagagaaca tctacaacat gaagaagttc agcaagctgc ctatcgccga gaacacccgg
      541 aacatccggc acagcaccaa cgacctgtac tacgagcaca gcctgaacct ggaaaagccc
      601 aacgagcact tcaccctgca caacatcaag ctgatctgcc agcagatcta ccacaaccag
      661 cccaaccagc tgaacctgcg gatcctggcc gtgctgctga gcaccagcta caagcaggaa
      721 aacattgcca acatcttcca gaacctgtgc aacaacggcc tgagctgcct gttcaccatc
      781 cagctgagca agaacaaggc cttcgtggac cagaacaaga tcgtgcacct gaagatcatc
      841 gacaccgacg acctgaagcg gaacaagaac aacaacatca ggcacaagat caccgtgtac
      901 tacatctgcg aggacctgaa gtacaagaac cagaagcagc aggaaagcaa cgagttcaaa
      961 tacgtgttca attacgagca gctggccttc ctgaagtgcg agaactacct gaccgtgcag
     1021 gtcgagaagc tcgtgggcat caactacaac aaagagttcc acaacctgaa ccccgagtac
     1081 atcagcttct actgcagcca gagagtgatc tgcaacaccc acatcgagga tctgtacaaa
     1141 gaggacgaca agaagcggaa gaactacatc ttctacgagc tgacctaccg ctgtgtgcct
     1201 cccaaagtgt gcagcttctg cggcaacaat gccgcctgca agctggaaaa actgtataac
     1261 aaagagaaag agatcaccca gttctacaac aacgtcgagt acaacgatgc cggcgtgttc
     1321 gaccggctgc tgctgaacaa agaagaggaa gaggaacgca acaaagccag cttcgtggcc
     1381 aacctcgagt ataacgccat taactacgac atcgacgaca tgatcacagt gggcagcaac
     1441 acctaccaca tcctggaaat ctgcaagtgc agcaaggact acgtgcagga aggctcctcc
     1501 tgcgtcaaga aagagaagga caagatctac atccgctacg ccaagaaagg cgcgccgtcg
     1561 acctccatca cggcctataa gagtgagggg gagtcagcgg agttctcctt cccactcaac
     1621 cttggagagg aaagcctgca gggagagttg agatggaagg cagagaaggc tccttcttcc
     1681 cagtcctgga tcaccttctc cctaaagaac caaaaggtgt ctgtgcagaa gtctactagc
     1741 aaccccaagt tccagctgtc cgaaacgctc ccactcaccc ttcagatacc ccaggtctcc
     1801 cttcagtttg ctggttctgg caacctgacc ctgactctgg acagagggat actgtatcag
     1861 gaagtgaacc tggtggtgat gaaagtgact cagcccgaca gcaacacttt gacctgtgag
     1921 gtgatgggac ccacctcacc caagatgaga ctgatcttga agcaggagaa tcaggaggcc
     1981 agggtctcca ggcaggagaa agtgattcaa gtgcaggccc ctgaagcagg ggtgtggcaa
     2041 tgtctactga gtgaaggtga agaggtcaag atggactcca agatccaggt tttatccaaa
     2101 gggttgaatt ccggcggcgg ctccggaggc ggtaccggtg gcggcgacct agccccacag
     2161 atgcttcgag aactccagga gactaatgcg gcgctgcaag acgtgagaga gctcttgcga
     2221 cagcaggtca aggagatcac cttcctgaag aatacggtga tggaatgtga cgcctgcggt
     2281 ctagacagaa acctgcctcc actagcacct ctgggcccac acccagaaac gctggtgaaa
     2341 gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac
     2401 agcggtaaga tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt
     2461 aaagttctgc tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga gcaactcggt
     2521 cgccgcatac actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat
     2581 cttacggatg gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac
     2641 actgcggcca acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg
     2701 cacaacatgg gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc
     2761 ataccaaacg acgagcgtga caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa
     2821 ctattaactg gcgaactact tactctagct tcccggcaac aattaataga ctggatggag
     2881 gcggataaag ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct
     2941 gataaatctg gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat
     3001 ggtaagccct cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa
     3061 cgaaatagac agatcgctga gataggtgcc tcactgatta agcattgggg ccctgactac
     3121 aaagaccatg acggtgatta taaagatcat gacatcgatt acaaggatga cgatgacaag
     3181 ggccctcacc accaccatca ccactaatta attaaggatc ccccgacctc gacctctggc
     3241 taataaagga aatttatttt cattgcaata gtgtgttgga attttttgtg tctctcactc
     3301 ggaaggacat atgggagggc aaatcatttg gtcgagatcc ctcggagatc tctagctaga
     3361 ggatcgatcc ccgccccgga cgaactaaac ctgactacga catctctgcc ccttcttcgc
     3421 ggggcagtgc atgtaatccc ttcagttggt tggtacaact tgccaactgg gccctgttcc
     3481 acatgtgaca cgggggggga ccaaacacaa aggggttctc tgactgtagt tgacatcctt
     3541 ataaatggat gtgcacattt gccaacactg agtggctttc atcctggagc agactttgca
     3601 gtctgtggac tgcaacacaa cattgccttt atgtgtaact cttggctgaa gctcttacac
     3661 caatgctggg ggacatgtac ctcccagggg cccaggaaga ctacgggagg ctacaccaac
     3721 gtcaatcaga ggggcctgtg tagctaccga taagcggacc ctcaagaggg cattagcaat
     3781 agtgtttata aggccccctt gttaacccta aacgggtagc atatgcttcc cgggtagtag
     3841 tatatactat ccagactaac cctaattcaa tagcatatgt tacccaacgg gaagcatatg
     3901 ctatcgaatt agggttagta aaagggtcct aaggaacagc gatatctccc accccatgag
     3961 ctgtcacggt tttatttaca tggggtcagg attccacgag ggtagtgaac cattttagtc
     4021 acaagggcag tggctgaaga tcaaggagcg ggcagtgaac tctcctgaat cttcgcctgc
     4081 ttcttcattc tccttcgttt agctaataga ataactgctg agttgtgaac agtaaggtgt
     4141 atgtgaggtg ctcgaaaaca aggtttcagg tgacgccccc agaataaaat ttggacgggg
     4201 ggttcagtgg tggcattgtg ctatgacacc aatataaccc tcacaaaccc cttgggcaat
     4261 aaatactagt gtaggaatga aacattctga atatctttaa caatagaaat ccatggggtg
     4321 gggacaagcc gtaaagactg gatgtccatc tcacacgaat ttatggctat gggcaacaca
     4381 taatcctagt gcaatatgat actggggtta ttaagatgtg tcccaggcag ggaccaagac
     4441 aggtgaacca tgttgttaca ctctatttgt aacaagggga aagagagtgg acgccgacag
     4501 cagcggactc cactggttgt ctctaacacc cccgaaaatt aaacggggct ccacgccaat
     4561 ggggcccata aacaaagaca agtggccact cttttttttg aaattgtgga gtgggggcac
     4621 gcgtcagccc ccacacgccg ccctgcggtt ttggactgta aaataagggt gtaataactt
     4681 ggctgattgt aaccccgcta accactgcgg tcaaaccact tgcccacaaa accactaatg
     4741 gcaccccggg gaatacctgc ataagtaggt gggcgggcca agataggggc gcgattgctg
     4801 cgatctggag gacaaattac acacacttgc gcctgagcgc caagcacagg gttgttggtc
     4861 ctcatattca cgaggtcgct gagagcacgg tgggctaatg ttgccatggg tagcatatac
     4921 tacccaaata tctggatagc atatgctatc ctaatctata tctgggtagc ataggctatc
     4981 ctaatctata tctgggtagc atatgctatc ctaatctata tctgggtagt atatgctatc
     5041 ctaatttata tctgggtagc ataggctatc ctaatctata tctgggtagc atatgctatc
     5101 ctaatctata tctgggtagt atatgctatc ctaatctgta tccgggtagc atatgctatc
     5161 ctaatagaga ttagggtagt atatgctatc ctaatttata tctgggtagc atatactacc
     5221 caaatatctg gatagcatat gctatcctaa tctatatctg ggtagcatat gctatcctaa
     5281 tctatatctg ggtagcatag gctatcctaa tctatatctg ggtagcatat gctatcctaa
     5341 tctatatctg ggtagtatat gctatcctaa tttatatctg ggtagcatag gctatcctaa
     5401 tctatatctg ggtagcatat gctatcctaa tctatatctg ggtagtatat gctatcctaa
     5461 tctgtatccg ggtagcatat gctatcctca tgataagctg tcaaacatga gaattttctt
     5521 gaagacgaaa gggcctcgtg atacgcctat ttttataggt taatgtcatg ataataatgg
     5581 tttcttagac gtcaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat
     5641 ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc
     5701 aataatattg aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc cttattccct
     5761 tttttgcggc attttgcctt cctgtttttg ctcacccaga aacgctggtg aaagtaaaag
     5821 atgctgaaga tcagttgggt gcacgagtgg gttacatcga actggatctc aacagcggta
     5881 agatccttga gagttttcgc cccgaagaac gttttccaat gatgagcact tttaaagttc
     5941 tgctatgtgg cgcggtatta tcccgtgttg acgccgggca agagcaactc ggtcgccgca
     6001 tacactattc tcagaatgac ttggttgagt actcaccagt cacagaaaag catcttacgg
     6061 atggcatgac agtaagagaa ttatgcagtg ctgccataac catgagtgat aacactgcgg
     6121 ccaacttact tctgacaacg atcggaggac cgaaggagct aaccgctttt ttgcacaaca
     6181 tgggggatca tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa gccataccaa
     6241 acgacgagcg tgacaccacg atgcctgcag caatggcaac aacgttgcgc aaactattaa
     6301 ctggcgaact acttactcta gcttcccggc aacaattaat agactggatg gaggcggata
     6361 aagttgcagg accacttctg cgctcggccc ttccggctgg ctggtttatt gctgataaat
     6421 ctggagccgg tgagcgtggg tctcgcggta tcattgcagc actggggcca gatggtaagc
     6481 cctcccgtat cgtagttatc tacacgacgg ggagtcaggc aactatggat gaacgaaata
     6541 gacagatcgc tgagataggt gcctcactga ttaagcattg gtaactgtca gaccaagttt
     6601 actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga
     6661 agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag
     6721 cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa
     6781 tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag
     6841 agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg
     6901 ttcttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat
     6961 acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta
     7021 ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg
     7081 gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc
     7141 gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa
     7201 gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc
     7261 tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt
     7321 caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct
     7381 tttgctggcc ttttgctcac atgttctttc ctgcgttatc ccctgattct gtggataacc
     7441 gtattaccgc ctttgagtga gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg
     7501 agtcagtgag cgaggaagcg gaagagcgcc caatacgcaa accgcctctc cccgcgcgtt
     7561 ggccgattca ttaatgcagc tggcacgaca ggtttcccga ctggaaagcg ggcagtgagc
     7621 gcaacgcaat taatgtgagt tagctcactc attaggcacc ccaggcttta cactttatgc
     7681 ttccggctcg tatgttgtgt ggaattgtga gcggataaca atttcacaca ggaaacagct
     7741 atgaccatga ttacgccaag ctctagctag aggtcgacca attctcatgt ttgacagctt
     7801 atcatcgcag atccgggcaa cgttgttgcc attgctgcag gcgcagaact ggtaggtatg
     7861 gaagatctat acattgaatc aatattggca attagccata ttagtcattg gttatatagc
     7921 ataaatcaat attggctatt ggccattgca tacgttgtat ctatatcata atatgtacat
     7981 ttatattggc tcatgtccaa tatgaccgcc atgttgacat tgattattga ctagttatta
     8041 atagtaatca attacggggt cattagttca tagcccatat atggagttcc gcgttacata
     8101 acttacggta aatggcccgc ctggctgacc gcccaacgac ccccgcccat tgacgtcaat
     8161 aatgacgtat gttcccatag taacgccaat agggactttc cattgacgtc aatgggtgga
     8221 gtatttacgg taaactgccc acttggcagt acatcaagtg tatcatatgc caagtccgcc
     8281 ccctattgac gtcaatgacg gtaaatggcc cgcctggcat tatgcccagt acatgacctt
     8341 acgggacttt cctacttggc agtacatcta cgtattagtc atcgctatta ccatggtgat
     8401 gcggttttgg cagtacacca atgggcgtgg atagcggttt gactcacggg gatttccaag
     8461 tctccacccc attgacgtca atgggagttt gttttggcac caaaatcaac gggactttcc
     8521 aaaatgtcgt aataaccccg ccccgttgac gcaaatgggc ggtaggcgtg tacggtggga
     8581 ggtctatata agcagagctc gtttagtgaa ccgtcagatc ctcactctct tccgcatcgc
     8641 tgtctgcgag ggccagctgt tgggctcgcg gttgaggaca aactcttcgc ggtctttcca
     8701 gtactcttgg atcggaaacc cgtcggcctc cgaacggtac tccgccaccg agggacctga
     8761 gcgagtccgc atcgaccgga tcggaaaacc t
//
