LOCUS       sequence-208743-        8176 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 8176)
  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 8176)
  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: P28-COMP-blac-flag-his
FEATURES             Location/Qualifiers
     source          1..8176
                     /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             1708..2493
                     /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             2500..2565
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             2572..2589
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    2627..2682
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(2681..2700)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(4918..4936)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5004..5108
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5109..5177,5178..5969)
                     /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      6140..6728
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6629..6648
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6882..6899
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    7016..7037
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7052..7057,7058..7075,7076..7082)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7090..7106
                     /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     7095..7117
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7114..7130
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7114..7130
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     enhancer        7401..7780
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        7781..7984
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     7935..7955
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     7981..8005
                     /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 ggccgccaga
      421 gtgaccgaga acaccatctg caaatacggc tacctgatcc agatgagcaa ccactacgag
      481 tgcaagtgca tcgagggcta cgtgctgatc aacgaggata cctgcggcaa gaaagtcgtg
      541 tgcgacaagg tggaaaacag cttcaaggcc tgcgacgagt acgcctactg cttcgacctg
      601 ggcaacaaga acaacgagaa gcagatcaag tgcatgtgcc ggaccgagta caccctgaca
      661 gccggcgtgt gcgtgcccaa cgtgtgccgg gacaaagtgt gcggcaaggg caagtgtatc
      721 gtggaccccg ccaacagcct gacccacacc tgtagctgca acatcggcac catcctgaac
      781 cagaacaagc tgtgcgacat ccagggcgac accccttgca gcctgaagtg cgccgagaac
      841 gaagtgtgca ccctggaagg caactactac acctgtaaag aggaccccag cagcaacggc
      901 ggaggcaaca ccgtggacca ggccgacaca agcggcgcgc cgtcgacctc catcacggcc
      961 tataagagtg agggggagtc agcggagttc tccttcccac tcaaccttgg agaggaaagc
     1021 ctgcagggag agttgagatg gaaggcagag aaggctcctt cttcccagtc ctggatcacc
     1081 ttctccctaa agaaccaaaa ggtgtctgtg cagaagtcta ctagcaaccc caagttccag
     1141 ctgtccgaaa cgctcccact cacccttcag ataccccagg tctcccttca gtttgctggt
     1201 tctggcaacc tgaccctgac tctggacaga gggatactgt atcaggaagt gaacctggtg
     1261 gtgatgaaag tgactcagcc cgacagcaac actttgacct gtgaggtgat gggacccacc
     1321 tcacccaaga tgagactgat cttgaagcag gagaatcagg aggccagggt ctccaggcag
     1381 gagaaagtga ttcaagtgca ggcccctgaa gcaggggtgt ggcaatgtct actgagtgaa
     1441 ggtgaagagg tcaagatgga ctccaagatc caggttttat ccaaagggtt gaattccggc
     1501 ggcggctccg gaggcggtac cggtggcggc gacctagccc cacagatgct tcgagaactc
     1561 caggagacta atgcggcgct gcaagacgtg agagagctct tgcgacagca ggtcaaggag
     1621 atcaccttcc tgaagaatac ggtgatggaa tgtgacgcct gcggtctaga cagaaacctg
     1681 cctccactag cacctctggg cccacaccca gaaacgctgg tgaaagtaaa agatgctgaa
     1741 gatcagttgg gtgcacgagt gggttacatc gaactggatc tcaacagcgg taagatcctt
     1801 gagagttttc gccccgaaga acgttttcca atgatgagca cttttaaagt tctgctatgt
     1861 ggcgcggtat tatcccgtat tgacgccggg caagagcaac tcggtcgccg catacactat
     1921 tctcagaatg acttggttga gtactcacca gtcacagaaa agcatcttac ggatggcatg
     1981 acagtaagag aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta
     2041 cttctgacaa cgatcggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat
     2101 catgtaactc gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag
     2161 cgtgacacca cgatgcctgt agcaatggca acaacgttgc gcaaactatt aactggcgaa
     2221 ctacttactc tagcttcccg gcaacaatta atagactgga tggaggcgga taaagttgca
     2281 ggaccacttc tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc
     2341 ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt
     2401 atcgtagtta tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc
     2461 gctgagatag gtgcctcact gattaagcat tggggccctg actacaaaga ccatgacggt
     2521 gattataaag atcatgacat cgattacaag gatgacgatg acaagggccc tcaccaccac
     2581 catcaccact aattaattaa ggatcccccg acctcgacct ctggctaata aaggaaattt
     2641 attttcattg caatagtgtg ttggaatttt ttgtgtctct cactcggaag gacatatggg
     2701 agggcaaatc atttggtcga gatccctcgg agatctctag ctagaggatc gatccccgcc
     2761 ccggacgaac taaacctgac tacgacatct ctgccccttc ttcgcggggc agtgcatgta
     2821 atcccttcag ttggttggta caacttgcca actgggccct gttccacatg tgacacgggg
     2881 ggggaccaaa cacaaagggg ttctctgact gtagttgaca tccttataaa tggatgtgca
     2941 catttgccaa cactgagtgg ctttcatcct ggagcagact ttgcagtctg tggactgcaa
     3001 cacaacattg cctttatgtg taactcttgg ctgaagctct tacaccaatg ctgggggaca
     3061 tgtacctccc aggggcccag gaagactacg ggaggctaca ccaacgtcaa tcagaggggc
     3121 ctgtgtagct accgataagc ggaccctcaa gagggcatta gcaatagtgt ttataaggcc
     3181 cccttgttaa ccctaaacgg gtagcatatg cttcccgggt agtagtatat actatccaga
     3241 ctaaccctaa ttcaatagca tatgttaccc aacgggaagc atatgctatc gaattagggt
     3301 tagtaaaagg gtcctaagga acagcgatat ctcccacccc atgagctgtc acggttttat
     3361 ttacatgggg tcaggattcc acgagggtag tgaaccattt tagtcacaag ggcagtggct
     3421 gaagatcaag gagcgggcag tgaactctcc tgaatcttcg cctgcttctt cattctcctt
     3481 cgtttagcta atagaataac tgctgagttg tgaacagtaa ggtgtatgtg aggtgctcga
     3541 aaacaaggtt tcaggtgacg cccccagaat aaaatttgga cggggggttc agtggtggca
     3601 ttgtgctatg acaccaatat aaccctcaca aaccccttgg gcaataaata ctagtgtagg
     3661 aatgaaacat tctgaatatc tttaacaata gaaatccatg gggtggggac aagccgtaaa
     3721 gactggatgt ccatctcaca cgaatttatg gctatgggca acacataatc ctagtgcaat
     3781 atgatactgg ggttattaag atgtgtccca ggcagggacc aagacaggtg aaccatgttg
     3841 ttacactcta tttgtaacaa ggggaaagag agtggacgcc gacagcagcg gactccactg
     3901 gttgtctcta acacccccga aaattaaacg gggctccacg ccaatggggc ccataaacaa
     3961 agacaagtgg ccactctttt ttttgaaatt gtggagtggg ggcacgcgtc agcccccaca
     4021 cgccgccctg cggttttgga ctgtaaaata agggtgtaat aacttggctg attgtaaccc
     4081 cgctaaccac tgcggtcaaa ccacttgccc acaaaaccac taatggcacc ccggggaata
     4141 cctgcataag taggtgggcg ggccaagata ggggcgcgat tgctgcgatc tggaggacaa
     4201 attacacaca cttgcgcctg agcgccaagc acagggttgt tggtcctcat attcacgagg
     4261 tcgctgagag cacggtgggc taatgttgcc atgggtagca tatactaccc aaatatctgg
     4321 atagcatatg ctatcctaat ctatatctgg gtagcatagg ctatcctaat ctatatctgg
     4381 gtagcatatg ctatcctaat ctatatctgg gtagtatatg ctatcctaat ttatatctgg
     4441 gtagcatagg ctatcctaat ctatatctgg gtagcatatg ctatcctaat ctatatctgg
     4501 gtagtatatg ctatcctaat ctgtatccgg gtagcatatg ctatcctaat agagattagg
     4561 gtagtatatg ctatcctaat ttatatctgg gtagcatata ctacccaaat atctggatag
     4621 catatgctat cctaatctat atctgggtag catatgctat cctaatctat atctgggtag
     4681 cataggctat cctaatctat atctgggtag catatgctat cctaatctat atctgggtag
     4741 tatatgctat cctaatttat atctgggtag cataggctat cctaatctat atctgggtag
     4801 catatgctat cctaatctat atctgggtag tatatgctat cctaatctgt atccgggtag
     4861 catatgctat cctcatgata agctgtcaaa catgagaatt ttcttgaaga cgaaagggcc
     4921 tcgtgatacg cctattttta taggttaatg tcatgataat aatggtttct tagacgtcag
     4981 gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt
     5041 caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa
     5101 ggaagagtat gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt
     5161 gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt
     5221 tgggtgcacg agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt
     5281 ttcgccccga agaacgtttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg
     5341 tattatcccg tgttgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga
     5401 atgacttggt tgagtactca ccagtcacag aaaagcatct tacggatggc atgacagtaa
     5461 gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga
     5521 caacgatcgg aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa
     5581 ctcgccttga tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca
     5641 ccacgatgcc tgcagcaatg gcaacaacgt tgcgcaaact attaactggc gaactactta
     5701 ctctagcttc ccggcaacaa ttaatagact ggatggaggc ggataaagtt gcaggaccac
     5761 ttctgcgctc ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc
     5821 gtgggtctcg cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag
     5881 ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga
     5941 taggtgcctc actgattaag cattggtaac tgtcagacca agtttactca tatatacttt
     6001 agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata
     6061 atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag
     6121 aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa
     6181 caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt
     6241 ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc
     6301 cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa
     6361 tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa
     6421 gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc
     6481 ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa
     6541 gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa
     6601 caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg
     6661 ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc
     6721 tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg
     6781 ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg
     6841 agtgagctga taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg
     6901 aagcggaaga gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat
     6961 gcagctggca cgacaggttt cccgactgga aagcgggcag tgagcgcaac gcaattaatg
     7021 tgagttagct cactcattag gcaccccagg ctttacactt tatgcttccg gctcgtatgt
     7081 tgtgtggaat tgtgagcgga taacaatttc acacaggaaa cagctatgac catgattacg
     7141 ccaagctcta gctagaggtc gaccaattct catgtttgac agcttatcat cgcagatccg
     7201 ggcaacgttg ttgccattgc tgcaggcgca gaactggtag gtatggaaga tctatacatt
     7261 gaatcaatat tggcaattag ccatattagt cattggttat atagcataaa tcaatattgg
     7321 ctattggcca ttgcatacgt tgtatctata tcataatatg tacatttata ttggctcatg
     7381 tccaatatga ccgccatgtt gacattgatt attgactagt tattaatagt aatcaattac
     7441 ggggtcatta gttcatagcc catatatgga gttccgcgtt acataactta cggtaaatgg
     7501 cccgcctggc tgaccgccca acgacccccg cccattgacg tcaataatga cgtatgttcc
     7561 catagtaacg ccaataggga ctttccattg acgtcaatgg gtggagtatt tacggtaaac
     7621 tgcccacttg gcagtacatc aagtgtatca tatgccaagt ccgcccccta ttgacgtcaa
     7681 tgacggtaaa tggcccgcct ggcattatgc ccagtacatg accttacggg actttcctac
     7741 ttggcagtac atctacgtat tagtcatcgc tattaccatg gtgatgcggt tttggcagta
     7801 caccaatggg cgtggatagc ggtttgactc acggggattt ccaagtctcc accccattga
     7861 cgtcaatggg agtttgtttt ggcaccaaaa tcaacgggac tttccaaaat gtcgtaataa
     7921 ccccgccccg ttgacgcaaa tgggcggtag gcgtgtacgg tgggaggtct atataagcag
     7981 agctcgttta gtgaaccgtc agatcctcac tctcttccgc atcgctgtct gcgagggcca
     8041 gctgttgggc tcgcggttga ggacaaactc ttcgcggtct ttccagtact cttggatcgg
     8101 aaacccgtcg gcctccgaac ggtactccgc caccgaggga cctgagcgag tccgcatcga
     8161 ccggatcgga aaacct
//
