LOCUS       sequence-224233-        8936 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 8936)
  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 8936)
  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_1463900-COMP-blac-flag-his
FEATURES             Location/Qualifiers
     source          1..8936
                     /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             2470..3255
                     /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             3262..3327
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             3334..3351
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    3387..3442
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(3441..3460)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(5678..5696)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5764..5868
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5869..5937,5938..6729)
                     /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      6900..7488
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7389..7408
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7642..7659
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    7776..7797
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7812..7817,7818..7835,7836..7842)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7850..7866
                     /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     7855..7877
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7874..7890
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7874..7890
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     enhancer        8161..8540
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        8541..8744
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     8695..8715
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     8741..8765
                     /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 ggccgccaac
      421 gagaacaact tcttcatcaa gatcctgcgg agccaggaaa agaagatcga gaacgaagtc
      481 tccctgtacg agaagctgaa caaccggatc ctgttcgagg gcagcgacga cttcagcaac
      541 ctggacaagc tgaagtccct gctgagcttc cagaagaaga ttgacaacac catccgcaag
      601 aagcagaaag aggaaaacaa gctcaagaag ttccagcagg ccatcaagaa cggcaacaac
      661 atctaccccg tgggcaacaa ccctcagggc ttcgagaaga aagagcgcga ggaaaaagag
      721 caggaaaaaa cctacatgaa caacgtgatc aacaagatca acgagcagaa gggcaactac
      781 tacgagccca acaagaacat caacaacaag tacaccgagg acctgaccag caacggcctg
      841 ctgtacaaca aggataagat caagaaccag ttcaacaatc cccaccagga aaaattcatc
      901 cggaagcacg agcacgagag agagcacgag ggcaagaacg aaggcaaaaa cgagcacaag
      961 aatgagcata agaacgaaca caaaaatgaa cacgtccacg tgcacgagca taacagcaag
     1021 aactatgagg gcaaccctcc caagaacgag taccccaaca tcaacttcgc caacatcctg
     1081 aacagcatcg acttctacaa gaaaatcgac ctggtcaaga actacttcgg ctacaacaac
     1141 aacgaggccc tgaaccagaa catgaaggaa cagaaaaaca tgcagaacca ccacgtggaa
     1201 aacgagaaga acaacatgat ccaggccaac aaccaggtgg acgagaatca gaacaatcac
     1261 cagtataaca acaacttcca caaccagaag aacgacaacc tgaagcagga caccaaggac
     1321 aacagcgccg agcagaccta caacaatgcc aacaacaata tcgaggacaa caatcacatc
     1381 aagttcttca tctccaagga catcgagaaa aaaaagaaca agcagaacga catcaatgag
     1441 gccgtgtaca acgccatcaa aaacaagaac ctgcacctcc ggcggaacaa catcaaagaa
     1501 atcttcgacg tggacgacct cgtgaagaat atcaagagct tcctgggcat caagagcaac
     1561 atccacgagg ccttcgagaa ccagaagctg atcatcaaga attgcaacta cgagagcttc
     1621 ggccccgagc tgtgctccgt ggatgagaag gccaaagaaa tgctgtggaa gtatgagaag
     1681 aagaaaaact ccgccggcgc gccgtcgacc tccatcacgg cctataagag tgagggggag
     1741 tcagcggagt tctccttccc actcaacctt ggagaggaaa gcctgcaggg agagttgaga
     1801 tggaaggcag agaaggctcc ttcttcccag tcctggatca ccttctccct aaagaaccaa
     1861 aaggtgtctg tgcagaagtc tactagcaac cccaagttcc agctgtccga aacgctccca
     1921 ctcacccttc agatacccca ggtctccctt cagtttgctg gttctggcaa cctgaccctg
     1981 actctggaca gagggatact gtatcaggaa gtgaacctgg tggtgatgaa agtgactcag
     2041 cccgacagca acactttgac ctgtgaggtg atgggaccca cctcacccaa gatgagactg
     2101 atcttgaagc aggagaatca ggaggccagg gtctccaggc aggagaaagt gattcaagtg
     2161 caggcccctg aagcaggggt gtggcaatgt ctactgagtg aaggtgaaga ggtcaagatg
     2221 gactccaaga tccaggtttt atccaaaggg ttgaattccg gcggcggctc cggaggcggt
     2281 accggtggcg gcgacctagc cccacagatg cttcgagaac tccaggagac taatgcggcg
     2341 ctgcaagacg tgagagagct cttgcgacag caggtcaagg agatcacctt cctgaagaat
     2401 acggtgatgg aatgtgacgc ctgcggtcta gacagaaacc tgcctccact agcacctctg
     2461 ggcccacacc cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcacga
     2521 gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa
     2581 gaacgttttc caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt
     2641 attgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt
     2701 gagtactcac cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc
     2761 agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac aacgatcgga
     2821 ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat
     2881 cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct
     2941 gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc
     3001 cggcaacaat taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg
     3061 gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc
     3121 ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg
     3181 acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca
     3241 ctgattaagc attggggccc tgactacaaa gaccatgacg gtgattataa agatcatgac
     3301 atcgattaca aggatgacga tgacaagggc cctcaccacc accatcacca ctaataataa
     3361 ggatcccccg acctcgacct ctggctaata aaggaaattt attttcattg caatagtgtg
     3421 ttggaatttt ttgtgtctct cactcggaag gacatatggg agggcaaatc atttggtcga
     3481 gatccctcgg agatctctag ctagaggatc gatccccgcc ccggacgaac taaacctgac
     3541 tacgacatct ctgccccttc ttcgcggggc agtgcatgta atcccttcag ttggttggta
     3601 caacttgcca actgggccct gttccacatg tgacacgggg ggggaccaaa cacaaagggg
     3661 ttctctgact gtagttgaca tccttataaa tggatgtgca catttgccaa cactgagtgg
     3721 ctttcatcct ggagcagact ttgcagtctg tggactgcaa cacaacattg cctttatgtg
     3781 taactcttgg ctgaagctct tacaccaatg ctgggggaca tgtacctccc aggggcccag
     3841 gaagactacg ggaggctaca ccaacgtcaa tcagaggggc ctgtgtagct accgataagc
     3901 ggaccctcaa gagggcatta gcaatagtgt ttataaggcc cccttgttaa ccctaaacgg
     3961 gtagcatatg cttcccgggt agtagtatat actatccaga ctaaccctaa ttcaatagca
     4021 tatgttaccc aacgggaagc atatgctatc gaattagggt tagtaaaagg gtcctaagga
     4081 acagcgatat ctcccacccc atgagctgtc acggttttat ttacatgggg tcaggattcc
     4141 acgagggtag tgaaccattt tagtcacaag ggcagtggct gaagatcaag gagcgggcag
     4201 tgaactctcc tgaatcttcg cctgcttctt cattctcctt cgtttagcta atagaataac
     4261 tgctgagttg tgaacagtaa ggtgtatgtg aggtgctcga aaacaaggtt tcaggtgacg
     4321 cccccagaat aaaatttgga cggggggttc agtggtggca ttgtgctatg acaccaatat
     4381 aaccctcaca aaccccttgg gcaataaata ctagtgtagg aatgaaacat tctgaatatc
     4441 tttaacaata gaaatccatg gggtggggac aagccgtaaa gactggatgt ccatctcaca
     4501 cgaatttatg gctatgggca acacataatc ctagtgcaat atgatactgg ggttattaag
     4561 atgtgtccca ggcagggacc aagacaggtg aaccatgttg ttacactcta tttgtaacaa
     4621 ggggaaagag agtggacgcc gacagcagcg gactccactg gttgtctcta acacccccga
     4681 aaattaaacg gggctccacg ccaatggggc ccataaacaa agacaagtgg ccactctttt
     4741 ttttgaaatt gtggagtggg ggcacgcgtc agcccccaca cgccgccctg cggttttgga
     4801 ctgtaaaata agggtgtaat aacttggctg attgtaaccc cgctaaccac tgcggtcaaa
     4861 ccacttgccc acaaaaccac taatggcacc ccggggaata cctgcataag taggtgggcg
     4921 ggccaagata ggggcgcgat tgctgcgatc tggaggacaa attacacaca cttgcgcctg
     4981 agcgccaagc acagggttgt tggtcctcat attcacgagg tcgctgagag cacggtgggc
     5041 taatgttgcc atgggtagca tatactaccc aaatatctgg atagcatatg ctatcctaat
     5101 ctatatctgg gtagcatagg ctatcctaat ctatatctgg gtagcatatg ctatcctaat
     5161 ctatatctgg gtagtatatg ctatcctaat ttatatctgg gtagcatagg ctatcctaat
     5221 ctatatctgg gtagcatatg ctatcctaat ctatatctgg gtagtatatg ctatcctaat
     5281 ctgtatccgg gtagcatatg ctatcctaat agagattagg gtagtatatg ctatcctaat
     5341 ttatatctgg gtagcatata ctacccaaat atctggatag catatgctat cctaatctat
     5401 atctgggtag catatgctat cctaatctat atctgggtag cataggctat cctaatctat
     5461 atctgggtag catatgctat cctaatctat atctgggtag tatatgctat cctaatttat
     5521 atctgggtag cataggctat cctaatctat atctgggtag catatgctat cctaatctat
     5581 atctgggtag tatatgctat cctaatctgt atccgggtag catatgctat cctcatgata
     5641 agctgtcaaa catgagaatt ttcttgaaga cgaaagggcc tcgtgatacg cctattttta
     5701 taggttaatg tcatgataat aatggtttct tagacgtcag gtggcacttt tcggggaaat
     5761 gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg
     5821 agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtat gagtattcaa
     5881 catttccgtg tcgcccttat tccctttttt gcggcatttt gccttcctgt ttttgctcac
     5941 ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg agtgggttac
     6001 atcgaactgg atctcaacag cggtaagatc cttgagagtt ttcgccccga agaacgtttt
     6061 ccaatgatga gcacttttaa agttctgcta tgtggcgcgg tattatcccg tgttgacgcc
     6121 gggcaagagc aactcggtcg ccgcatacac tattctcaga atgacttggt tgagtactca
     6181 ccagtcacag aaaagcatct tacggatggc atgacagtaa gagaattatg cagtgctgcc
     6241 ataaccatga gtgataacac tgcggccaac ttacttctga caacgatcgg aggaccgaag
     6301 gagctaaccg cttttttgca caacatgggg gatcatgtaa ctcgccttga tcgttgggaa
     6361 ccggagctga atgaagccat accaaacgac gagcgtgaca ccacgatgcc tgcagcaatg
     6421 gcaacaacgt tgcgcaaact attaactggc gaactactta ctctagcttc ccggcaacaa
     6481 ttaatagact ggatggaggc ggataaagtt gcaggaccac ttctgcgctc ggcccttccg
     6541 gctggctggt ttattgctga taaatctgga gccggtgagc gtgggtctcg cggtatcatt
     6601 gcagcactgg ggccagatgg taagccctcc cgtatcgtag ttatctacac gacggggagt
     6661 caggcaacta tggatgaacg aaatagacag atcgctgaga taggtgcctc actgattaag
     6721 cattggtaac tgtcagacca agtttactca tatatacttt agattgattt aaaacttcat
     6781 ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac caaaatccct
     6841 taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct
     6901 tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca
     6961 gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc
     7021 agcagagcgc agataccaaa tactgttctt ctagtgtagc cgtagttagg ccaccacttc
     7081 aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct
     7141 gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag
     7201 gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc
     7261 tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg
     7321 agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag
     7381 cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt
     7441 gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac
     7501 gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt ctttcctgcg
     7561 ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga taccgctcgc
     7621 cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga gcgcccaata
     7681 cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctggca cgacaggttt
     7741 cccgactgga aagcgggcag tgagcgcaac gcaattaatg tgagttagct cactcattag
     7801 gcaccccagg ctttacactt tatgcttccg gctcgtatgt tgtgtggaat tgtgagcgga
     7861 taacaatttc acacaggaaa cagctatgac catgattacg ccaagctcta gctagaggtc
     7921 gaccaattct catgtttgac agcttatcat cgcagatccg ggcaacgttg ttgccattgc
     7981 tgcaggcgca gaactggtag gtatggaaga tctatacatt gaatcaatat tggcaattag
     8041 ccatattagt cattggttat atagcataaa tcaatattgg ctattggcca ttgcatacgt
     8101 tgtatctata tcataatatg tacatttata ttggctcatg tccaatatga ccgccatgtt
     8161 gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc
     8221 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
     8281 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
     8341 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
     8401 aagtgtatca tatgccaagt ccgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
     8461 ggcattatgc ccagtacatg accttacggg actttcctac ttggcagtac atctacgtat
     8521 tagtcatcgc tattaccatg gtgatgcggt tttggcagta caccaatggg cgtggatagc
     8581 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt
     8641 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaataa ccccgccccg ttgacgcaaa
     8701 tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctcgttta gtgaaccgtc
     8761 agatcctcac tctcttccgc atcgctgtct gcgagggcca gctgttgggc tcgcggttga
     8821 ggacaaactc ttcgcggtct ttccagtact cttggatcgg aaacccgtcg gcctccgaac
     8881 ggtactccgc caccgaggga cctgagcgag tccgcatcga ccggatcgga aaacct
//
