LOCUS       sequence-224544-        9794 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 9794)
  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 9794)
  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..9794
                     /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    4245..4300
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(4299..4318)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(6536..6554)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6622..6726
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6727..6795,6796..7587)
                     /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      7758..8346
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8247..8266
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     8500..8517
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    8634..8655
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(8670..8675,8676..8693,8694..8700)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8708..8724
                     /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     8713..8735
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     8732..8748
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     8732..8748
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     enhancer        9019..9398
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        9399..9602
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     9553..9573
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     9599..9623
                     /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 aataataagg atcccccgac ctcgacctct ggctaataaa ggaaatttat
     4261 tttcattgca atagtgtgtt ggaatttttt gtgtctctca ctcggaagga catatgggag
     4321 ggcaaatcat ttggtcgaga tccctcggag atctctagct agaggatcga tccccgcccc
     4381 ggacgaacta aacctgacta cgacatctct gccccttctt cgcggggcag tgcatgtaat
     4441 cccttcagtt ggttggtaca acttgccaac tgggccctgt tccacatgtg acacgggggg
     4501 ggaccaaaca caaaggggtt ctctgactgt agttgacatc cttataaatg gatgtgcaca
     4561 tttgccaaca ctgagtggct ttcatcctgg agcagacttt gcagtctgtg gactgcaaca
     4621 caacattgcc tttatgtgta actcttggct gaagctctta caccaatgct gggggacatg
     4681 tacctcccag gggcccagga agactacggg aggctacacc aacgtcaatc agaggggcct
     4741 gtgtagctac cgataagcgg accctcaaga gggcattagc aatagtgttt ataaggcccc
     4801 cttgttaacc ctaaacgggt agcatatgct tcccgggtag tagtatatac tatccagact
     4861 aaccctaatt caatagcata tgttacccaa cgggaagcat atgctatcga attagggtta
     4921 gtaaaagggt cctaaggaac agcgatatct cccaccccat gagctgtcac ggttttattt
     4981 acatggggtc aggattccac gagggtagtg aaccatttta gtcacaaggg cagtggctga
     5041 agatcaagga gcgggcagtg aactctcctg aatcttcgcc tgcttcttca ttctccttcg
     5101 tttagctaat agaataactg ctgagttgtg aacagtaagg tgtatgtgag gtgctcgaaa
     5161 acaaggtttc aggtgacgcc cccagaataa aatttggacg gggggttcag tggtggcatt
     5221 gtgctatgac accaatataa ccctcacaaa ccccttgggc aataaatact agtgtaggaa
     5281 tgaaacattc tgaatatctt taacaataga aatccatggg gtggggacaa gccgtaaaga
     5341 ctggatgtcc atctcacacg aatttatggc tatgggcaac acataatcct agtgcaatat
     5401 gatactgggg ttattaagat gtgtcccagg cagggaccaa gacaggtgaa ccatgttgtt
     5461 acactctatt tgtaacaagg ggaaagagag tggacgccga cagcagcgga ctccactggt
     5521 tgtctctaac acccccgaaa attaaacggg gctccacgcc aatggggccc ataaacaaag
     5581 acaagtggcc actctttttt ttgaaattgt ggagtggggg cacgcgtcag cccccacacg
     5641 ccgccctgcg gttttggact gtaaaataag ggtgtaataa cttggctgat tgtaaccccg
     5701 ctaaccactg cggtcaaacc acttgcccac aaaaccacta atggcacccc ggggaatacc
     5761 tgcataagta ggtgggcggg ccaagatagg ggcgcgattg ctgcgatctg gaggacaaat
     5821 tacacacact tgcgcctgag cgccaagcac agggttgttg gtcctcatat tcacgaggtc
     5881 gctgagagca cggtgggcta atgttgccat gggtagcata tactacccaa atatctggat
     5941 agcatatgct atcctaatct atatctgggt agcataggct atcctaatct atatctgggt
     6001 agcatatgct atcctaatct atatctgggt agtatatgct atcctaattt atatctgggt
     6061 agcataggct atcctaatct atatctgggt agcatatgct atcctaatct atatctgggt
     6121 agtatatgct atcctaatct gtatccgggt agcatatgct atcctaatag agattagggt
     6181 agtatatgct atcctaattt atatctgggt agcatatact acccaaatat ctggatagca
     6241 tatgctatcc taatctatat ctgggtagca tatgctatcc taatctatat ctgggtagca
     6301 taggctatcc taatctatat ctgggtagca tatgctatcc taatctatat ctgggtagta
     6361 tatgctatcc taatttatat ctgggtagca taggctatcc taatctatat ctgggtagca
     6421 tatgctatcc taatctatat ctgggtagta tatgctatcc taatctgtat ccgggtagca
     6481 tatgctatcc tcatgataag ctgtcaaaca tgagaatttt cttgaagacg aaagggcctc
     6541 gtgatacgcc tatttttata ggttaatgtc atgataataa tggtttctta gacgtcaggt
     6601 ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca
     6661 aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg
     6721 aagagtatga gtattcaaca tttccgtgtc gcccttattc ccttttttgc ggcattttgc
     6781 cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga agatcagttg
     6841 ggtgcacgag tgggttacat cgaactggat ctcaacagcg gtaagatcct tgagagtttt
     6901 cgccccgaag aacgttttcc aatgatgagc acttttaaag ttctgctatg tggcgcggta
     6961 ttatcccgtg ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat
     7021 gacttggttg agtactcacc agtcacagaa aagcatctta cggatggcat gacagtaaga
     7081 gaattatgca gtgctgccat aaccatgagt gataacactg cggccaactt acttctgaca
     7141 acgatcggag gaccgaagga gctaaccgct tttttgcaca acatggggga tcatgtaact
     7201 cgccttgatc gttgggaacc ggagctgaat gaagccatac caaacgacga gcgtgacacc
     7261 acgatgcctg cagcaatggc aacaacgttg cgcaaactat taactggcga actacttact
     7321 ctagcttccc ggcaacaatt aatagactgg atggaggcgg ataaagttgc aggaccactt
     7381 ctgcgctcgg cccttccggc tggctggttt attgctgata aatctggagc cggtgagcgt
     7441 gggtctcgcg gtatcattgc agcactgggg ccagatggta agccctcccg tatcgtagtt
     7501 atctacacga cggggagtca ggcaactatg gatgaacgaa atagacagat cgctgagata
     7561 ggtgcctcac tgattaagca ttggtaactg tcagaccaag tttactcata tatactttag
     7621 attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat
     7681 ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa
     7741 aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca
     7801 aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt
     7861 ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct agtgtagccg
     7921 tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc
     7981 ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga
     8041 cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc
     8101 agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc
     8161 gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca
     8221 ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg
     8281 tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta
     8341 tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct
     8401 cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac cgcctttgag
     8461 tgagctgata ccgctcgccg cagccgaacg accgagcgca gcgagtcagt gagcgaggaa
     8521 gcggaagagc gcccaatacg caaaccgcct ctccccgcgc gttggccgat tcattaatgc
     8581 agctggcacg acaggtttcc cgactggaaa gcgggcagtg agcgcaacgc aattaatgtg
     8641 agttagctca ctcattaggc accccaggct ttacacttta tgcttccggc tcgtatgttg
     8701 tgtggaattg tgagcggata acaatttcac acaggaaaca gctatgacca tgattacgcc
     8761 aagctctagc tagaggtcga ccaattctca tgtttgacag cttatcatcg cagatccggg
     8821 caacgttgtt gccattgctg caggcgcaga actggtaggt atggaagatc tatacattga
     8881 atcaatattg gcaattagcc atattagtca ttggttatat agcataaatc aatattggct
     8941 attggccatt gcatacgttg tatctatatc ataatatgta catttatatt ggctcatgtc
     9001 caatatgacc gccatgttga cattgattat tgactagtta ttaatagtaa tcaattacgg
     9061 ggtcattagt tcatagccca tatatggagt tccgcgttac ataacttacg gtaaatggcc
     9121 cgcctggctg accgcccaac gacccccgcc cattgacgtc aataatgacg tatgttccca
     9181 tagtaacgcc aatagggact ttccattgac gtcaatgggt ggagtattta cggtaaactg
     9241 cccacttggc agtacatcaa gtgtatcata tgccaagtcc gccccctatt gacgtcaatg
     9301 acggtaaatg gcccgcctgg cattatgccc agtacatgac cttacgggac tttcctactt
     9361 ggcagtacat ctacgtatta gtcatcgcta ttaccatggt gatgcggttt tggcagtaca
     9421 ccaatgggcg tggatagcgg tttgactcac ggggatttcc aagtctccac cccattgacg
     9481 tcaatgggag tttgttttgg caccaaaatc aacgggactt tccaaaatgt cgtaataacc
     9541 ccgccccgtt gacgcaaatg ggcggtaggc gtgtacggtg ggaggtctat ataagcagag
     9601 ctcgtttagt gaaccgtcag atcctcactc tcttccgcat cgctgtctgc gagggccagc
     9661 tgttgggctc gcggttgagg acaaactctt cgcggtcttt ccagtactct tggatcggaa
     9721 acccgtcggc ctccgaacgg tactccgcca ccgagggacc tgagcgagtc cgcatcgacc
     9781 ggatcggaaa acct
//
