LOCUS       sequence-208756-        9796 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 9796)
  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 9796)
  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: P47-COMP-blac-flag-his
FEATURES             Location/Qualifiers
     source          1..9796
                     /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             3328..4113
                     /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             4120..4185
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             4192..4209
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    4247..4302
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(4301..4320)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(6538..6556)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6624..6728
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6729..6797,6798..7589)
                     /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      7760..8348
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8249..8268
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     8502..8519
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    8636..8657
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(8672..8677,8678..8695,8696..8702)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8710..8726
                     /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     8715..8737
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     8734..8750
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     8734..8750
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     enhancer        9021..9400
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        9401..9604
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     9555..9575
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     9601..9625
                     /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 ggccgccttc
      421 tacttcgagc agaccgagga actgctgatc aagaacgccc acagcgacat ctgcaagaga
      481 gagcactaca tcagcttcaa caaagagatc atcaagctct gcctgttcag ccagcccagc
      541 ggcgtgcaca gcaagtacaa cctgatcgtg aacaccctga aagaggaaaa gtgggaagag
      601 aagtacaaga tcctcaaaga gaacaccttc aagatcctga agtactttta caccttcgtg
      661 aacaagaacg agctggtcat catcttttgc tttaacaagc tgatcaacaa ggccccttac
      721 gagtgccacc ggtccattaa caaggtggac ggcaagggca acgaggaaca gagagtgcgg
      781 atcaactaca accacatcga ccctctgagc ctggtggact acaccagcaa taccttcgac
      841 ttcctgggca ccagctacct gctgatctgc ggcatcaacg agaaccggaa gctgagcccc
      901 ggcagccacg tgttcatcct gtgccgggcc agcaaggaca gcggcatcac ctggggcctg
      961 accttcagct tctactaccc tcacgccaag agcagcgtgc tgtactccaa ggtggtgccc
     1021 aagatcagct acaacgagat cggcttcctg ttcttcagcc ccgacgtgtc catcaacaag
     1081 tacatcaagt gcacctaccg cgagggcaac cacttcgact gcgacgacat ccacttcatc
     1141 gacgacaagt acctgctgga catcaccaaa gtgcggggct actacctgac catcctgagc
     1201 aacaagaagg acaagaaaga aaaagtgaag ctgtggtaca tctacaacag cgtgatcagc
     1261 gagagcatgc ggaacaacgc catcagcagc ggcaatgagt acagagaggg caatctgttc
     1321 ctgctgaacg acgccacagt gctgtacaac tacctgaacg agaaggactc ctacctgtac
     1381 ctgatcaagc acaagggcag cgtgaagcac tgcaccctgc tgtatctgaa gaaagagtac
     1441 gccaaccccg gcttcgtgaa agagattaag agaaaccacg ccaccaccct gaccaaggac
     1501 aacagcaaca actaccgcta cgtgtgcaac atcaactacg acgacctgag catcaacctc
     1561 aacgaccggt acttcatcgt gtccgtgtac aacggcgtga aaaccgacat ccggtcctgc
     1621 ttctacttta gctacgacaa catcctcgag cccgtgaaca ttatcaacaa gatcgtgaag
     1681 gtctacgagt actccaacta cgccatctac gccttccaca ttaacaaaga cctcgagagc
     1741 tacttcctgt ttaacaagaa aatggaatgc ttcctcggcg acaacttcta catctacctc
     1801 aacatcaatt tcatcaacac ctacaagctg gacaactcct tcgacaacaa caatgagaag
     1861 gtggtcaacc tgtaccccaa caacctcatc tactacaagc tccccaacat taactacaag
     1921 aataccttcc tgtccaacgt gcacttccct ccctacacca aatacgtgca gctcaacgac
     1981 aaagagtaca tctttaagct gcccagctac atcgaagagg acatccacac cgagctgcac
     2041 tttctccaga aagtcaacaa gttctacaag aagaaaacca tcttccacaa gggcggccac
     2101 aacaacgacc tggtgctggg catcgacttc agcaaaaaga ccaacatctg cagctacggc
     2161 tacagcaaga ccagctgcga gtacacacac atcgacatca acaaaatcaa tatcaaggtg
     2221 cccatgaacg acgagatcac ctccgacatc accctggcca tcatctgccc cgtgtccaag
     2281 aaaaaccaga acgtgtgctt ccaccacacc tacgacaatg gcaagaaggt gctgatccgg
     2341 gaccacctgt acgaccggaa cggctacctg aatgtgtacc ctaagatcta tatctacacc
     2401 cctaagctga accagatcta cgaggaatcc aagctcgtga tccggaaaga gttcatcgag
     2461 atcttcaaga acaacaagtg caattacaag aacgtgatcc tgtgcaagtg ctacgccaac
     2521 aacaaatatt acgatatcac cttcaatctg atcggcgcgc cgtcgacctc catcacggcc
     2581 tataagagtg agggggagtc agcggagttc tccttcccac tcaaccttgg agaggaaagc
     2641 ctgcagggag agttgagatg gaaggcagag aaggctcctt cttcccagtc ctggatcacc
     2701 ttctccctaa agaaccaaaa ggtgtctgtg cagaagtcta ctagcaaccc caagttccag
     2761 ctgtccgaaa cgctcccact cacccttcag ataccccagg tctcccttca gtttgctggt
     2821 tctggcaacc tgaccctgac tctggacaga gggatactgt atcaggaagt gaacctggtg
     2881 gtgatgaaag tgactcagcc cgacagcaac actttgacct gtgaggtgat gggacccacc
     2941 tcacccaaga tgagactgat cttgaagcag gagaatcagg aggccagggt ctccaggcag
     3001 gagaaagtga ttcaagtgca ggcccctgaa gcaggggtgt ggcaatgtct actgagtgaa
     3061 ggtgaagagg tcaagatgga ctccaagatc caggttttat ccaaagggtt gaattccggc
     3121 ggcggctccg gaggcggtac cggtggcggc gacctagccc cacagatgct tcgagaactc
     3181 caggagacta atgcggcgct gcaagacgtg agagagctct tgcgacagca ggtcaaggag
     3241 atcaccttcc tgaagaatac ggtgatggaa tgtgacgcct gcggtctaga cagaaacctg
     3301 cctccactag cacctctggg cccacaccca gaaacgctgg tgaaagtaaa agatgctgaa
     3361 gatcagttgg gtgcacgagt gggttacatc gaactggatc tcaacagcgg taagatcctt
     3421 gagagttttc gccccgaaga acgttttcca atgatgagca cttttaaagt tctgctatgt
     3481 ggcgcggtat tatcccgtat tgacgccggg caagagcaac tcggtcgccg catacactat
     3541 tctcagaatg acttggttga gtactcacca gtcacagaaa agcatcttac ggatggcatg
     3601 acagtaagag aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta
     3661 cttctgacaa cgatcggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat
     3721 catgtaactc gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag
     3781 cgtgacacca cgatgcctgt agcaatggca acaacgttgc gcaaactatt aactggcgaa
     3841 ctacttactc tagcttcccg gcaacaatta atagactgga tggaggcgga taaagttgca
     3901 ggaccacttc tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc
     3961 ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt
     4021 atcgtagtta tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc
     4081 gctgagatag gtgcctcact gattaagcat tggggccctg actacaaaga ccatgacggt
     4141 gattataaag atcatgacat cgattacaag gatgacgatg acaagggccc tcaccaccac
     4201 catcaccact aattaattaa ggatcccccg acctcgacct ctggctaata aaggaaattt
     4261 attttcattg caatagtgtg ttggaatttt ttgtgtctct cactcggaag gacatatggg
     4321 agggcaaatc atttggtcga gatccctcgg agatctctag ctagaggatc gatccccgcc
     4381 ccggacgaac taaacctgac tacgacatct ctgccccttc ttcgcggggc agtgcatgta
     4441 atcccttcag ttggttggta caacttgcca actgggccct gttccacatg tgacacgggg
     4501 ggggaccaaa cacaaagggg ttctctgact gtagttgaca tccttataaa tggatgtgca
     4561 catttgccaa cactgagtgg ctttcatcct ggagcagact ttgcagtctg tggactgcaa
     4621 cacaacattg cctttatgtg taactcttgg ctgaagctct tacaccaatg ctgggggaca
     4681 tgtacctccc aggggcccag gaagactacg ggaggctaca ccaacgtcaa tcagaggggc
     4741 ctgtgtagct accgataagc ggaccctcaa gagggcatta gcaatagtgt ttataaggcc
     4801 cccttgttaa ccctaaacgg gtagcatatg cttcccgggt agtagtatat actatccaga
     4861 ctaaccctaa ttcaatagca tatgttaccc aacgggaagc atatgctatc gaattagggt
     4921 tagtaaaagg gtcctaagga acagcgatat ctcccacccc atgagctgtc acggttttat
     4981 ttacatgggg tcaggattcc acgagggtag tgaaccattt tagtcacaag ggcagtggct
     5041 gaagatcaag gagcgggcag tgaactctcc tgaatcttcg cctgcttctt cattctcctt
     5101 cgtttagcta atagaataac tgctgagttg tgaacagtaa ggtgtatgtg aggtgctcga
     5161 aaacaaggtt tcaggtgacg cccccagaat aaaatttgga cggggggttc agtggtggca
     5221 ttgtgctatg acaccaatat aaccctcaca aaccccttgg gcaataaata ctagtgtagg
     5281 aatgaaacat tctgaatatc tttaacaata gaaatccatg gggtggggac aagccgtaaa
     5341 gactggatgt ccatctcaca cgaatttatg gctatgggca acacataatc ctagtgcaat
     5401 atgatactgg ggttattaag atgtgtccca ggcagggacc aagacaggtg aaccatgttg
     5461 ttacactcta tttgtaacaa ggggaaagag agtggacgcc gacagcagcg gactccactg
     5521 gttgtctcta acacccccga aaattaaacg gggctccacg ccaatggggc ccataaacaa
     5581 agacaagtgg ccactctttt ttttgaaatt gtggagtggg ggcacgcgtc agcccccaca
     5641 cgccgccctg cggttttgga ctgtaaaata agggtgtaat aacttggctg attgtaaccc
     5701 cgctaaccac tgcggtcaaa ccacttgccc acaaaaccac taatggcacc ccggggaata
     5761 cctgcataag taggtgggcg ggccaagata ggggcgcgat tgctgcgatc tggaggacaa
     5821 attacacaca cttgcgcctg agcgccaagc acagggttgt tggtcctcat attcacgagg
     5881 tcgctgagag cacggtgggc taatgttgcc atgggtagca tatactaccc aaatatctgg
     5941 atagcatatg ctatcctaat ctatatctgg gtagcatagg ctatcctaat ctatatctgg
     6001 gtagcatatg ctatcctaat ctatatctgg gtagtatatg ctatcctaat ttatatctgg
     6061 gtagcatagg ctatcctaat ctatatctgg gtagcatatg ctatcctaat ctatatctgg
     6121 gtagtatatg ctatcctaat ctgtatccgg gtagcatatg ctatcctaat agagattagg
     6181 gtagtatatg ctatcctaat ttatatctgg gtagcatata ctacccaaat atctggatag
     6241 catatgctat cctaatctat atctgggtag catatgctat cctaatctat atctgggtag
     6301 cataggctat cctaatctat atctgggtag catatgctat cctaatctat atctgggtag
     6361 tatatgctat cctaatttat atctgggtag cataggctat cctaatctat atctgggtag
     6421 catatgctat cctaatctat atctgggtag tatatgctat cctaatctgt atccgggtag
     6481 catatgctat cctcatgata agctgtcaaa catgagaatt ttcttgaaga cgaaagggcc
     6541 tcgtgatacg cctattttta taggttaatg tcatgataat aatggtttct tagacgtcag
     6601 gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt
     6661 caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa
     6721 ggaagagtat gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt
     6781 gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt
     6841 tgggtgcacg agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt
     6901 ttcgccccga agaacgtttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg
     6961 tattatcccg tgttgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga
     7021 atgacttggt tgagtactca ccagtcacag aaaagcatct tacggatggc atgacagtaa
     7081 gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga
     7141 caacgatcgg aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa
     7201 ctcgccttga tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca
     7261 ccacgatgcc tgcagcaatg gcaacaacgt tgcgcaaact attaactggc gaactactta
     7321 ctctagcttc ccggcaacaa ttaatagact ggatggaggc ggataaagtt gcaggaccac
     7381 ttctgcgctc ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc
     7441 gtgggtctcg cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag
     7501 ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga
     7561 taggtgcctc actgattaag cattggtaac tgtcagacca agtttactca tatatacttt
     7621 agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata
     7681 atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag
     7741 aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa
     7801 caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt
     7861 ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc
     7921 cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa
     7981 tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa
     8041 gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc
     8101 ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa
     8161 gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa
     8221 caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg
     8281 ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc
     8341 tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg
     8401 ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg
     8461 agtgagctga taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg
     8521 aagcggaaga gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat
     8581 gcagctggca cgacaggttt cccgactgga aagcgggcag tgagcgcaac gcaattaatg
     8641 tgagttagct cactcattag gcaccccagg ctttacactt tatgcttccg gctcgtatgt
     8701 tgtgtggaat tgtgagcgga taacaatttc acacaggaaa cagctatgac catgattacg
     8761 ccaagctcta gctagaggtc gaccaattct catgtttgac agcttatcat cgcagatccg
     8821 ggcaacgttg ttgccattgc tgcaggcgca gaactggtag gtatggaaga tctatacatt
     8881 gaatcaatat tggcaattag ccatattagt cattggttat atagcataaa tcaatattgg
     8941 ctattggcca ttgcatacgt tgtatctata tcataatatg tacatttata ttggctcatg
     9001 tccaatatga ccgccatgtt gacattgatt attgactagt tattaatagt aatcaattac
     9061 ggggtcatta gttcatagcc catatatgga gttccgcgtt acataactta cggtaaatgg
     9121 cccgcctggc tgaccgccca acgacccccg cccattgacg tcaataatga cgtatgttcc
     9181 catagtaacg ccaataggga ctttccattg acgtcaatgg gtggagtatt tacggtaaac
     9241 tgcccacttg gcagtacatc aagtgtatca tatgccaagt ccgcccccta ttgacgtcaa
     9301 tgacggtaaa tggcccgcct ggcattatgc ccagtacatg accttacggg actttcctac
     9361 ttggcagtac atctacgtat tagtcatcgc tattaccatg gtgatgcggt tttggcagta
     9421 caccaatggg cgtggatagc ggtttgactc acggggattt ccaagtctcc accccattga
     9481 cgtcaatggg agtttgtttt ggcaccaaaa tcaacgggac tttccaaaat gtcgtaataa
     9541 ccccgccccg ttgacgcaaa tgggcggtag gcgtgtacgg tgggaggtct atataagcag
     9601 agctcgttta gtgaaccgtc agatcctcac tctcttccgc atcgctgtct gcgagggcca
     9661 gctgttgggc tcgcggttga ggacaaactc ttcgcggtct ttccagtact cttggatcgg
     9721 aaacccgtcg gcctccgaac ggtactccgc caccgaggga cctgagcgag tccgcatcga
     9781 ccggatcgga aaacct
//
