LOCUS       sequence-224481-        8171 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 8171)
  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 8171)
  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: P25-COMP-blac-flag-his
FEATURES             Location/Qualifiers
     source          1..8171
                     /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             1705..2490
                     /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             2497..2562
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             2569..2586
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    2622..2677
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(2676..2695)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(4913..4931)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4999..5103
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5104..5172,5173..5964)
                     /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      6135..6723
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6624..6643
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6877..6894
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    7011..7032
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7047..7052,7053..7070,7071..7077)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7085..7101
                     /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     7090..7112
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7109..7125
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7109..7125
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     enhancer        7396..7775
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        7776..7979
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     7930..7950
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     7976..8000
                     /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 ggccgccaaa
      421 gtgaccgtgg acaccgtgtg caagcggggc ttcctgatcc agatgagcgg ccacctggaa
      481 tgcaagtgcg agaacgacct ggtgctggtc aacgaggaaa cctgcgaaga gaaggtgctg
      541 aagtgcgacg agaaaaccgt gaacaagccc tgcggcgact tcagcaagtg catcaagatc
      601 gacggcaacc ccgtgtccta cgcctgcaag tgcaacctgg gctacgacat ggtcaacaac
      661 gtgtgcatcc ccaacgagtg caagaacgtg gcctgcggca acggcaagtg tatcctggac
      721 accagcaacc ctgtgaaaac cggcgtgtgc tcctgcaaca tcggcaaggt gcccaacgtg
      781 caggaccaga acaagtgcag caaggacggc gagacaaagt gcagcctgaa gtgcctgaaa
      841 gagaatgagg cctgcaaggc cgtggacggc atctacaagt gcgactgcaa ggatggcttc
      901 atcatcgaca acgaggccag catctgcacc ggcgcgccgt cgacctccat cacggcctat
      961 aagagtgagg gggagtcagc ggagttctcc ttcccactca accttggaga ggaaagcctg
     1021 cagggagagt tgagatggaa ggcagagaag gctccttctt cccagtcctg gatcaccttc
     1081 tccctaaaga accaaaaggt gtctgtgcag aagtctacta gcaaccccaa gttccagctg
     1141 tccgaaacgc tcccactcac ccttcagata ccccaggtct cccttcagtt tgctggttct
     1201 ggcaacctga ccctgactct ggacagaggg atactgtatc aggaagtgaa cctggtggtg
     1261 atgaaagtga ctcagcccga cagcaacact ttgacctgtg aggtgatggg acccacctca
     1321 cccaagatga gactgatctt gaagcaggag aatcaggagg ccagggtctc caggcaggag
     1381 aaagtgattc aagtgcaggc ccctgaagca ggggtgtggc aatgtctact gagtgaaggt
     1441 gaagaggtca agatggactc caagatccag gttttatcca aagggttgaa ttccggcggc
     1501 ggctccggag gcggtaccgg tggcggcgac ctagccccac agatgcttcg agaactccag
     1561 gagactaatg cggcgctgca agacgtgaga gagctcttgc gacagcaggt caaggagatc
     1621 accttcctga agaatacggt gatggaatgt gacgcctgcg gtctagacag aaacctgcct
     1681 ccactagcac ctctgggccc acacccagaa acgctggtga aagtaaaaga tgctgaagat
     1741 cagttgggtg cacgagtggg ttacatcgaa ctggatctca acagcggtaa gatccttgag
     1801 agttttcgcc ccgaagaacg ttttccaatg atgagcactt ttaaagttct gctatgtggc
     1861 gcggtattat cccgtattga cgccgggcaa gagcaactcg gtcgccgcat acactattct
     1921 cagaatgact tggttgagta ctcaccagtc acagaaaagc atcttacgga tggcatgaca
     1981 gtaagagaat tatgcagtgc tgccataacc atgagtgata acactgcggc caacttactt
     2041 ctgacaacga tcggaggacc gaaggagcta accgcttttt tgcacaacat gggggatcat
     2101 gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt
     2161 gacaccacga tgcctgtagc aatggcaaca acgttgcgca aactattaac tggcgaacta
     2221 cttactctag cttcccggca acaattaata gactggatgg aggcggataa agttgcagga
     2281 ccacttctgc gctcggccct tccggctggc tggtttattg ctgataaatc tggagccggt
     2341 gagcgtgggt ctcgcggtat cattgcagca ctggggccag atggtaagcc ctcccgtatc
     2401 gtagttatct acacgacggg gagtcaggca actatggatg aacgaaatag acagatcgct
     2461 gagataggtg cctcactgat taagcattgg ggccctgact acaaagacca tgacggtgat
     2521 tataaagatc atgacatcga ttacaaggat gacgatgaca agggccctca ccaccaccat
     2581 caccactaat aataaggatc ccccgacctc gacctctggc taataaagga aatttatttt
     2641 cattgcaata gtgtgttgga attttttgtg tctctcactc ggaaggacat atgggagggc
     2701 aaatcatttg gtcgagatcc ctcggagatc tctagctaga ggatcgatcc ccgccccgga
     2761 cgaactaaac ctgactacga catctctgcc ccttcttcgc ggggcagtgc atgtaatccc
     2821 ttcagttggt tggtacaact tgccaactgg gccctgttcc acatgtgaca cgggggggga
     2881 ccaaacacaa aggggttctc tgactgtagt tgacatcctt ataaatggat gtgcacattt
     2941 gccaacactg agtggctttc atcctggagc agactttgca gtctgtggac tgcaacacaa
     3001 cattgccttt atgtgtaact cttggctgaa gctcttacac caatgctggg ggacatgtac
     3061 ctcccagggg cccaggaaga ctacgggagg ctacaccaac gtcaatcaga ggggcctgtg
     3121 tagctaccga taagcggacc ctcaagaggg cattagcaat agtgtttata aggccccctt
     3181 gttaacccta aacgggtagc atatgcttcc cgggtagtag tatatactat ccagactaac
     3241 cctaattcaa tagcatatgt tacccaacgg gaagcatatg ctatcgaatt agggttagta
     3301 aaagggtcct aaggaacagc gatatctccc accccatgag ctgtcacggt tttatttaca
     3361 tggggtcagg attccacgag ggtagtgaac cattttagtc acaagggcag tggctgaaga
     3421 tcaaggagcg ggcagtgaac tctcctgaat cttcgcctgc ttcttcattc tccttcgttt
     3481 agctaataga ataactgctg agttgtgaac agtaaggtgt atgtgaggtg ctcgaaaaca
     3541 aggtttcagg tgacgccccc agaataaaat ttggacgggg ggttcagtgg tggcattgtg
     3601 ctatgacacc aatataaccc tcacaaaccc cttgggcaat aaatactagt gtaggaatga
     3661 aacattctga atatctttaa caatagaaat ccatggggtg gggacaagcc gtaaagactg
     3721 gatgtccatc tcacacgaat ttatggctat gggcaacaca taatcctagt gcaatatgat
     3781 actggggtta ttaagatgtg tcccaggcag ggaccaagac aggtgaacca tgttgttaca
     3841 ctctatttgt aacaagggga aagagagtgg acgccgacag cagcggactc cactggttgt
     3901 ctctaacacc cccgaaaatt aaacggggct ccacgccaat ggggcccata aacaaagaca
     3961 agtggccact cttttttttg aaattgtgga gtgggggcac gcgtcagccc ccacacgccg
     4021 ccctgcggtt ttggactgta aaataagggt gtaataactt ggctgattgt aaccccgcta
     4081 accactgcgg tcaaaccact tgcccacaaa accactaatg gcaccccggg gaatacctgc
     4141 ataagtaggt gggcgggcca agataggggc gcgattgctg cgatctggag gacaaattac
     4201 acacacttgc gcctgagcgc caagcacagg gttgttggtc ctcatattca cgaggtcgct
     4261 gagagcacgg tgggctaatg ttgccatggg tagcatatac tacccaaata tctggatagc
     4321 atatgctatc ctaatctata tctgggtagc ataggctatc ctaatctata tctgggtagc
     4381 atatgctatc ctaatctata tctgggtagt atatgctatc ctaatttata tctgggtagc
     4441 ataggctatc ctaatctata tctgggtagc atatgctatc ctaatctata tctgggtagt
     4501 atatgctatc ctaatctgta tccgggtagc atatgctatc ctaatagaga ttagggtagt
     4561 atatgctatc ctaatttata tctgggtagc atatactacc caaatatctg gatagcatat
     4621 gctatcctaa tctatatctg ggtagcatat gctatcctaa tctatatctg ggtagcatag
     4681 gctatcctaa tctatatctg ggtagcatat gctatcctaa tctatatctg ggtagtatat
     4741 gctatcctaa tttatatctg ggtagcatag gctatcctaa tctatatctg ggtagcatat
     4801 gctatcctaa tctatatctg ggtagtatat gctatcctaa tctgtatccg ggtagcatat
     4861 gctatcctca tgataagctg tcaaacatga gaattttctt gaagacgaaa gggcctcgtg
     4921 atacgcctat ttttataggt taatgtcatg ataataatgg tttcttagac gtcaggtggc
     4981 acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat
     5041 atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag
     5101 agtatgagta ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt
     5161 cctgtttttg ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt
     5221 gcacgagtgg gttacatcga actggatctc aacagcggta agatccttga gagttttcgc
     5281 cccgaagaac gttttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta
     5341 tcccgtgttg acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac
     5401 ttggttgagt actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa
     5461 ttatgcagtg ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg
     5521 atcggaggac cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc
     5581 cttgatcgtt gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg
     5641 atgcctgcag caatggcaac aacgttgcgc aaactattaa ctggcgaact acttactcta
     5701 gcttcccggc aacaattaat agactggatg gaggcggata aagttgcagg accacttctg
     5761 cgctcggccc ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggg
     5821 tctcgcggta tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc
     5881 tacacgacgg ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt
     5941 gcctcactga ttaagcattg gtaactgtca gaccaagttt actcatatat actttagatt
     6001 gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc
     6061 atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag
     6121 atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa
     6181 aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg
     6241 aaggtaactg gcttcagcag agcgcagata ccaaatactg ttcttctagt gtagccgtag
     6301 ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg
     6361 ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga
     6421 tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc
     6481 ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc
     6541 acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga
     6601 gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt
     6661 cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg
     6721 aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac
     6781 atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc ctttgagtga
     6841 gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg
     6901 gaagagcgcc caatacgcaa accgcctctc cccgcgcgtt ggccgattca ttaatgcagc
     6961 tggcacgaca ggtttcccga ctggaaagcg ggcagtgagc gcaacgcaat taatgtgagt
     7021 tagctcactc attaggcacc ccaggcttta cactttatgc ttccggctcg tatgttgtgt
     7081 ggaattgtga gcggataaca atttcacaca ggaaacagct atgaccatga ttacgccaag
     7141 ctctagctag aggtcgacca attctcatgt ttgacagctt atcatcgcag atccgggcaa
     7201 cgttgttgcc attgctgcag gcgcagaact ggtaggtatg gaagatctat acattgaatc
     7261 aatattggca attagccata ttagtcattg gttatatagc ataaatcaat attggctatt
     7321 ggccattgca tacgttgtat ctatatcata atatgtacat ttatattggc tcatgtccaa
     7381 tatgaccgcc atgttgacat tgattattga ctagttatta atagtaatca attacggggt
     7441 cattagttca tagcccatat atggagttcc gcgttacata acttacggta aatggcccgc
     7501 ctggctgacc gcccaacgac ccccgcccat tgacgtcaat aatgacgtat gttcccatag
     7561 taacgccaat agggactttc cattgacgtc aatgggtgga gtatttacgg taaactgccc
     7621 acttggcagt acatcaagtg tatcatatgc caagtccgcc ccctattgac gtcaatgacg
     7681 gtaaatggcc cgcctggcat tatgcccagt acatgacctt acgggacttt cctacttggc
     7741 agtacatcta cgtattagtc atcgctatta ccatggtgat gcggttttgg cagtacacca
     7801 atgggcgtgg atagcggttt gactcacggg gatttccaag tctccacccc attgacgtca
     7861 atgggagttt gttttggcac caaaatcaac gggactttcc aaaatgtcgt aataaccccg
     7921 ccccgttgac gcaaatgggc ggtaggcgtg tacggtggga ggtctatata agcagagctc
     7981 gtttagtgaa ccgtcagatc ctcactctct tccgcatcgc tgtctgcgag ggccagctgt
     8041 tgggctcgcg gttgaggaca aactcttcgc ggtctttcca gtactcttgg atcggaaacc
     8101 cgtcggcctc cgaacggtac tccgccaccg agggacctga gcgagtccgc atcgaccgga
     8161 tcggaaaacc t
//
