LOCUS       sequence-208815-        8344 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 8344)
  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 8344)
  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: GEST-COMP-blac-flag-his
FEATURES             Location/Qualifiers
     source          1..8344
                     /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             1876..2661
                     /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             2668..2733
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             2740..2757
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    2795..2850
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(2849..2868)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(5086..5104)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5172..5276
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5277..5345,5346..6137)
                     /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      6308..6896
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6797..6816
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7050..7067
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    7184..7205
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7220..7225,7226..7243,7244..7250)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7258..7274
                     /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     7263..7285
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7282..7298
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7282..7298
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     enhancer        7569..7948
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        7949..8152
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     8103..8123
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     8149..8173
                     /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 ggccgccatt
      421 cagctgcaca gcgtgaacta cgcccctctg aactacatcg agatcggcga caccctgagc
      481 aacaaagtgg gcgagctgtg gcacagccac ctgaacagct tcatcgacat cgtgtccacc
      541 aaggtggtca acaagctgga agatgacctg tccaacggca acaacctcga gaagttcatg
      601 atcctgctcg aggacgacgc cgacatcttc gacgccagcg cctacgaggg caagaacctg
      661 gccctgatcc agaccgagtt catccggaag ctgaaggaca agttcaagaa cagcaagttc
      721 ggccagaagc tgaaaaagct gggcagcaag gccaaagaga agctgctgag cctgtaccag
      781 aagcacaagg gcaagctgcg gcacttcttc agcatcctgc tgaagtccct ggtcatccct
      841 atcgccgtgc agttcatcag aaagaacctg aacaagtgga agcagcggac cctggaagcc
      901 acacagaagc tggacgagca gagcaagaat atcgcccagc ccatcatcaa ccggctgtac
      961 aacagcttcg aggacaagat cgaagagtac agccaggaaa acaagatcaa cgtcgaggat
     1021 gagctgaacg ccctgggcca gctggacaag gacaagcagg acatccagaa actcgaggaa
     1081 caggaaaagg ctctgctgca gggcgcgccg tcgacctcca tcacggccta taagagtgag
     1141 ggggagtcag cggagttctc cttcccactc aaccttggag aggaaagcct gcagggagag
     1201 ttgagatgga aggcagagaa ggctccttct tcccagtcct ggatcacctt ctccctaaag
     1261 aaccaaaagg tgtctgtgca gaagtctact agcaacccca agttccagct gtccgaaacg
     1321 ctcccactca cccttcagat accccaggtc tcccttcagt ttgctggttc tggcaacctg
     1381 accctgactc tggacagagg gatactgtat caggaagtga acctggtggt gatgaaagtg
     1441 actcagcccg acagcaacac tttgacctgt gaggtgatgg gacccacctc acccaagatg
     1501 agactgatct tgaagcagga gaatcaggag gccagggtct ccaggcagga gaaagtgatt
     1561 caagtgcagg cccctgaagc aggggtgtgg caatgtctac tgagtgaagg tgaagaggtc
     1621 aagatggact ccaagatcca ggttttatcc aaagggttga attccggcgg cggctccgga
     1681 ggcggtaccg gtggcggcga cctagcccca cagatgcttc gagaactcca ggagactaat
     1741 gcggcgctgc aagacgtgag agagctcttg cgacagcagg tcaaggagat caccttcctg
     1801 aagaatacgg tgatggaatg tgacgcctgc ggtctagaca gaaacctgcc tccactagca
     1861 cctctgggcc cacacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt
     1921 gcacgagtgg gttacatcga actggatctc aacagcggta agatccttga gagttttcgc
     1981 cccgaagaac gttttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta
     2041 tcccgtattg acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac
     2101 ttggttgagt actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa
     2161 ttatgcagtg ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg
     2221 atcggaggac cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc
     2281 cttgatcgtt gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg
     2341 atgcctgtag caatggcaac aacgttgcgc aaactattaa ctggcgaact acttactcta
     2401 gcttcccggc aacaattaat agactggatg gaggcggata aagttgcagg accacttctg
     2461 cgctcggccc ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggg
     2521 tctcgcggta tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc
     2581 tacacgacgg ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt
     2641 gcctcactga ttaagcattg gggccctgac tacaaagacc atgacggtga ttataaagat
     2701 catgacatcg attacaagga tgacgatgac aagggccctc accaccacca tcaccactaa
     2761 ttaattaagg atcccccgac ctcgacctct ggctaataaa ggaaatttat tttcattgca
     2821 atagtgtgtt ggaatttttt gtgtctctca ctcggaagga catatgggag ggcaaatcat
     2881 ttggtcgaga tccctcggag atctctagct agaggatcga tccccgcccc ggacgaacta
     2941 aacctgacta cgacatctct gccccttctt cgcggggcag tgcatgtaat cccttcagtt
     3001 ggttggtaca acttgccaac tgggccctgt tccacatgtg acacgggggg ggaccaaaca
     3061 caaaggggtt ctctgactgt agttgacatc cttataaatg gatgtgcaca tttgccaaca
     3121 ctgagtggct ttcatcctgg agcagacttt gcagtctgtg gactgcaaca caacattgcc
     3181 tttatgtgta actcttggct gaagctctta caccaatgct gggggacatg tacctcccag
     3241 gggcccagga agactacggg aggctacacc aacgtcaatc agaggggcct gtgtagctac
     3301 cgataagcgg accctcaaga gggcattagc aatagtgttt ataaggcccc cttgttaacc
     3361 ctaaacgggt agcatatgct tcccgggtag tagtatatac tatccagact aaccctaatt
     3421 caatagcata tgttacccaa cgggaagcat atgctatcga attagggtta gtaaaagggt
     3481 cctaaggaac agcgatatct cccaccccat gagctgtcac ggttttattt acatggggtc
     3541 aggattccac gagggtagtg aaccatttta gtcacaaggg cagtggctga agatcaagga
     3601 gcgggcagtg aactctcctg aatcttcgcc tgcttcttca ttctccttcg tttagctaat
     3661 agaataactg ctgagttgtg aacagtaagg tgtatgtgag gtgctcgaaa acaaggtttc
     3721 aggtgacgcc cccagaataa aatttggacg gggggttcag tggtggcatt gtgctatgac
     3781 accaatataa ccctcacaaa ccccttgggc aataaatact agtgtaggaa tgaaacattc
     3841 tgaatatctt taacaataga aatccatggg gtggggacaa gccgtaaaga ctggatgtcc
     3901 atctcacacg aatttatggc tatgggcaac acataatcct agtgcaatat gatactgggg
     3961 ttattaagat gtgtcccagg cagggaccaa gacaggtgaa ccatgttgtt acactctatt
     4021 tgtaacaagg ggaaagagag tggacgccga cagcagcgga ctccactggt tgtctctaac
     4081 acccccgaaa attaaacggg gctccacgcc aatggggccc ataaacaaag acaagtggcc
     4141 actctttttt ttgaaattgt ggagtggggg cacgcgtcag cccccacacg ccgccctgcg
     4201 gttttggact gtaaaataag ggtgtaataa cttggctgat tgtaaccccg ctaaccactg
     4261 cggtcaaacc acttgcccac aaaaccacta atggcacccc ggggaatacc tgcataagta
     4321 ggtgggcggg ccaagatagg ggcgcgattg ctgcgatctg gaggacaaat tacacacact
     4381 tgcgcctgag cgccaagcac agggttgttg gtcctcatat tcacgaggtc gctgagagca
     4441 cggtgggcta atgttgccat gggtagcata tactacccaa atatctggat agcatatgct
     4501 atcctaatct atatctgggt agcataggct atcctaatct atatctgggt agcatatgct
     4561 atcctaatct atatctgggt agtatatgct atcctaattt atatctgggt agcataggct
     4621 atcctaatct atatctgggt agcatatgct atcctaatct atatctgggt agtatatgct
     4681 atcctaatct gtatccgggt agcatatgct atcctaatag agattagggt agtatatgct
     4741 atcctaattt atatctgggt agcatatact acccaaatat ctggatagca tatgctatcc
     4801 taatctatat ctgggtagca tatgctatcc taatctatat ctgggtagca taggctatcc
     4861 taatctatat ctgggtagca tatgctatcc taatctatat ctgggtagta tatgctatcc
     4921 taatttatat ctgggtagca taggctatcc taatctatat ctgggtagca tatgctatcc
     4981 taatctatat ctgggtagta tatgctatcc taatctgtat ccgggtagca tatgctatcc
     5041 tcatgataag ctgtcaaaca tgagaatttt cttgaagacg aaagggcctc gtgatacgcc
     5101 tatttttata ggttaatgtc atgataataa tggtttctta gacgtcaggt ggcacttttc
     5161 ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc
     5221 cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtatga
     5281 gtattcaaca tttccgtgtc gcccttattc ccttttttgc ggcattttgc cttcctgttt
     5341 ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga agatcagttg ggtgcacgag
     5401 tgggttacat cgaactggat ctcaacagcg gtaagatcct tgagagtttt cgccccgaag
     5461 aacgttttcc aatgatgagc acttttaaag ttctgctatg tggcgcggta ttatcccgtg
     5521 ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat gacttggttg
     5581 agtactcacc agtcacagaa aagcatctta cggatggcat gacagtaaga gaattatgca
     5641 gtgctgccat aaccatgagt gataacactg cggccaactt acttctgaca acgatcggag
     5701 gaccgaagga gctaaccgct tttttgcaca acatggggga tcatgtaact cgccttgatc
     5761 gttgggaacc ggagctgaat gaagccatac caaacgacga gcgtgacacc acgatgcctg
     5821 cagcaatggc aacaacgttg cgcaaactat taactggcga actacttact ctagcttccc
     5881 ggcaacaatt aatagactgg atggaggcgg ataaagttgc aggaccactt ctgcgctcgg
     5941 cccttccggc tggctggttt attgctgata aatctggagc cggtgagcgt gggtctcgcg
     6001 gtatcattgc agcactgggg ccagatggta agccctcccg tatcgtagtt atctacacga
     6061 cggggagtca ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac
     6121 tgattaagca ttggtaactg tcagaccaag tttactcata tatactttag attgatttaa
     6181 aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca
     6241 aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag
     6301 gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac
     6361 cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa
     6421 ctggcttcag cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttaggcc
     6481 accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag
     6541 tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac
     6601 cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc
     6661 gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc
     6721 ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca
     6781 cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc
     6841 tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg
     6901 ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgttct
     6961 ttcctgcgtt atcccctgat tctgtggata accgtattac cgcctttgag tgagctgata
     7021 ccgctcgccg cagccgaacg accgagcgca gcgagtcagt gagcgaggaa gcggaagagc
     7081 gcccaatacg caaaccgcct ctccccgcgc gttggccgat tcattaatgc agctggcacg
     7141 acaggtttcc cgactggaaa gcgggcagtg agcgcaacgc aattaatgtg agttagctca
     7201 ctcattaggc accccaggct ttacacttta tgcttccggc tcgtatgttg tgtggaattg
     7261 tgagcggata acaatttcac acaggaaaca gctatgacca tgattacgcc aagctctagc
     7321 tagaggtcga ccaattctca tgtttgacag cttatcatcg cagatccggg caacgttgtt
     7381 gccattgctg caggcgcaga actggtaggt atggaagatc tatacattga atcaatattg
     7441 gcaattagcc atattagtca ttggttatat agcataaatc aatattggct attggccatt
     7501 gcatacgttg tatctatatc ataatatgta catttatatt ggctcatgtc caatatgacc
     7561 gccatgttga cattgattat tgactagtta ttaatagtaa tcaattacgg ggtcattagt
     7621 tcatagccca tatatggagt tccgcgttac ataacttacg gtaaatggcc cgcctggctg
     7681 accgcccaac gacccccgcc cattgacgtc aataatgacg tatgttccca tagtaacgcc
     7741 aatagggact ttccattgac gtcaatgggt ggagtattta cggtaaactg cccacttggc
     7801 agtacatcaa gtgtatcata tgccaagtcc gccccctatt gacgtcaatg acggtaaatg
     7861 gcccgcctgg cattatgccc agtacatgac cttacgggac tttcctactt ggcagtacat
     7921 ctacgtatta gtcatcgcta ttaccatggt gatgcggttt tggcagtaca ccaatgggcg
     7981 tggatagcgg tttgactcac ggggatttcc aagtctccac cccattgacg tcaatgggag
     8041 tttgttttgg caccaaaatc aacgggactt tccaaaatgt cgtaataacc ccgccccgtt
     8101 gacgcaaatg ggcggtaggc gtgtacggtg ggaggtctat ataagcagag ctcgtttagt
     8161 gaaccgtcag atcctcactc tcttccgcat cgctgtctgc gagggccagc tgttgggctc
     8221 gcggttgagg acaaactctt cgcggtcttt ccagtactct tggatcggaa acccgtcggc
     8281 ctccgaacgg tactccgcca ccgagggacc tgagcgagtc cgcatcgacc ggatcggaaa
     8341 acct
//
