LOCUS       sequence-208765-        8332 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 8332)
  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 8332)
  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: SPECT1-COMP-blac-flag-his
FEATURES             Location/Qualifiers
     source          1..8332
                     /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             1864..2649
                     /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             2656..2721
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             2728..2745
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    2783..2838
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(2837..2856)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(5074..5092)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5160..5264
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5265..5333,5334..6125)
                     /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      6296..6884
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6785..6804
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7038..7055
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    7172..7193
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7208..7213,7214..7231,7232..7238)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7246..7262
                     /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     7251..7273
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7270..7286
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7270..7286
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     enhancer        7557..7936
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        7937..8140
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     8091..8111
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     8137..8161
                     /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 ggccgcctac
      421 aacctgaaca acgacctgag caagaacaac aacttcgccc tgaacaccta cgtgcggaag
      481 gacgacgtgg aagatgacag caagaacgag atcgtggaca acatccagaa gatggtcgac
      541 gacttcagcg acgacatcgg cttcgtgaaa accagcatga gagaggtgct gctggacacc
      601 gaggccagcc tggaagaggt gtccgatcac gtggtgcaga atatcgccaa gtacagcctg
      661 accatcgagg aaaagctgaa cctgttcgac ggcctgctgg aagagttcat cgaaaacaac
      721 aagggcctga tcagcaacct ggccaagcgg cagcagaagc tgaagggcga caagatcaag
      781 aaagtgtgcg acctgatcct gaagaagctc aagaagctgg aaaacgtcaa caagctgatc
      841 aagtacaaga tcatcctgaa gtacggcaac aaggacaaca agaaagagat gatccagacc
      901 ctgaagaacg aggaaggcct gagcgacgac tttaagaaca atctggccaa ctacgagaca
      961 gagcagaaca acgatgacat caaagagatc gagctggtca acttcatcag caccaattac
     1021 gacaagttcg tggtcaacct cgaggacctc aacaaagagc tgctgaagga cctgaacatg
     1081 gccctgagcg gcgcgccgtc gacctccatc acggcctata agagtgaggg ggagtcagcg
     1141 gagttctcct tcccactcaa ccttggagag gaaagcctgc agggagagtt gagatggaag
     1201 gcagagaagg ctccttcttc ccagtcctgg atcaccttct ccctaaagaa ccaaaaggtg
     1261 tctgtgcaga agtctactag caaccccaag ttccagctgt ccgaaacgct cccactcacc
     1321 cttcagatac cccaggtctc ccttcagttt gctggttctg gcaacctgac cctgactctg
     1381 gacagaggga tactgtatca ggaagtgaac ctggtggtga tgaaagtgac tcagcccgac
     1441 agcaacactt tgacctgtga ggtgatggga cccacctcac ccaagatgag actgatcttg
     1501 aagcaggaga atcaggaggc cagggtctcc aggcaggaga aagtgattca agtgcaggcc
     1561 cctgaagcag gggtgtggca atgtctactg agtgaaggtg aagaggtcaa gatggactcc
     1621 aagatccagg ttttatccaa agggttgaat tccggcggcg gctccggagg cggtaccggt
     1681 ggcggcgacc tagccccaca gatgcttcga gaactccagg agactaatgc ggcgctgcaa
     1741 gacgtgagag agctcttgcg acagcaggtc aaggagatca ccttcctgaa gaatacggtg
     1801 atggaatgtg acgcctgcgg tctagacaga aacctgcctc cactagcacc tctgggccca
     1861 cacccagaaa cgctggtgaa agtaaaagat gctgaagatc agttgggtgc acgagtgggt
     1921 tacatcgaac tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgt
     1981 tttccaatga tgagcacttt taaagttctg ctatgtggcg cggtattatc ccgtattgac
     2041 gccgggcaag agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtac
     2101 tcaccagtca cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct
     2161 gccataacca tgagtgataa cactgcggcc aacttacttc tgacaacgat cggaggaccg
     2221 aaggagctaa ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg
     2281 gaaccggagc tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgtagca
     2341 atggcaacaa cgttgcgcaa actattaact ggcgaactac ttactctagc ttcccggcaa
     2401 caattaatag actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt
     2461 ccggctggct ggtttattgc tgataaatct ggagccggtg agcgtgggtc tcgcggtatc
     2521 attgcagcac tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg
     2581 agtcaggcaa ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt
     2641 aagcattggg gccctgacta caaagaccat gacggtgatt ataaagatca tgacatcgat
     2701 tacaaggatg acgatgacaa gggccctcac caccaccatc accactaatt aattaaggat
     2761 cccccgacct cgacctctgg ctaataaagg aaatttattt tcattgcaat agtgtgttgg
     2821 aattttttgt gtctctcact cggaaggaca tatgggaggg caaatcattt ggtcgagatc
     2881 cctcggagat ctctagctag aggatcgatc cccgccccgg acgaactaaa cctgactacg
     2941 acatctctgc cccttcttcg cggggcagtg catgtaatcc cttcagttgg ttggtacaac
     3001 ttgccaactg ggccctgttc cacatgtgac acgggggggg accaaacaca aaggggttct
     3061 ctgactgtag ttgacatcct tataaatgga tgtgcacatt tgccaacact gagtggcttt
     3121 catcctggag cagactttgc agtctgtgga ctgcaacaca acattgcctt tatgtgtaac
     3181 tcttggctga agctcttaca ccaatgctgg gggacatgta cctcccaggg gcccaggaag
     3241 actacgggag gctacaccaa cgtcaatcag aggggcctgt gtagctaccg ataagcggac
     3301 cctcaagagg gcattagcaa tagtgtttat aaggccccct tgttaaccct aaacgggtag
     3361 catatgcttc ccgggtagta gtatatacta tccagactaa ccctaattca atagcatatg
     3421 ttacccaacg ggaagcatat gctatcgaat tagggttagt aaaagggtcc taaggaacag
     3481 cgatatctcc caccccatga gctgtcacgg ttttatttac atggggtcag gattccacga
     3541 gggtagtgaa ccattttagt cacaagggca gtggctgaag atcaaggagc gggcagtgaa
     3601 ctctcctgaa tcttcgcctg cttcttcatt ctccttcgtt tagctaatag aataactgct
     3661 gagttgtgaa cagtaaggtg tatgtgaggt gctcgaaaac aaggtttcag gtgacgcccc
     3721 cagaataaaa tttggacggg gggttcagtg gtggcattgt gctatgacac caatataacc
     3781 ctcacaaacc ccttgggcaa taaatactag tgtaggaatg aaacattctg aatatcttta
     3841 acaatagaaa tccatggggt ggggacaagc cgtaaagact ggatgtccat ctcacacgaa
     3901 tttatggcta tgggcaacac ataatcctag tgcaatatga tactggggtt attaagatgt
     3961 gtcccaggca gggaccaaga caggtgaacc atgttgttac actctatttg taacaagggg
     4021 aaagagagtg gacgccgaca gcagcggact ccactggttg tctctaacac ccccgaaaat
     4081 taaacggggc tccacgccaa tggggcccat aaacaaagac aagtggccac tctttttttt
     4141 gaaattgtgg agtgggggca cgcgtcagcc cccacacgcc gccctgcggt tttggactgt
     4201 aaaataaggg tgtaataact tggctgattg taaccccgct aaccactgcg gtcaaaccac
     4261 ttgcccacaa aaccactaat ggcaccccgg ggaatacctg cataagtagg tgggcgggcc
     4321 aagatagggg cgcgattgct gcgatctgga ggacaaatta cacacacttg cgcctgagcg
     4381 ccaagcacag ggttgttggt cctcatattc acgaggtcgc tgagagcacg gtgggctaat
     4441 gttgccatgg gtagcatata ctacccaaat atctggatag catatgctat cctaatctat
     4501 atctgggtag cataggctat cctaatctat atctgggtag catatgctat cctaatctat
     4561 atctgggtag tatatgctat cctaatttat atctgggtag cataggctat cctaatctat
     4621 atctgggtag catatgctat cctaatctat atctgggtag tatatgctat cctaatctgt
     4681 atccgggtag catatgctat cctaatagag attagggtag tatatgctat cctaatttat
     4741 atctgggtag catatactac ccaaatatct ggatagcata tgctatccta atctatatct
     4801 gggtagcata tgctatccta atctatatct gggtagcata ggctatccta atctatatct
     4861 gggtagcata tgctatccta atctatatct gggtagtata tgctatccta atttatatct
     4921 gggtagcata ggctatccta atctatatct gggtagcata tgctatccta atctatatct
     4981 gggtagtata tgctatccta atctgtatcc gggtagcata tgctatcctc atgataagct
     5041 gtcaaacatg agaattttct tgaagacgaa agggcctcgt gatacgccta tttttatagg
     5101 ttaatgtcat gataataatg gtttcttaga cgtcaggtgg cacttttcgg ggaaatgtgc
     5161 gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac
     5221 aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtatgagt attcaacatt
     5281 tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt gctcacccag
     5341 aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg ggttacatcg
     5401 aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa cgttttccaa
     5461 tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtgtt gacgccgggc
     5521 aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag tactcaccag
     5581 tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt gctgccataa
     5641 ccatgagtga taacactgcg gccaacttac ttctgacaac gatcggagga ccgaaggagc
     5701 taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt tgggaaccgg
     5761 agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgca gcaatggcaa
     5821 caacgttgcg caaactatta actggcgaac tacttactct agcttcccgg caacaattaa
     5881 tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc cttccggctg
     5941 gctggtttat tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt atcattgcag
     6001 cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg gggagtcagg
     6061 caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg attaagcatt
     6121 ggtaactgtc agaccaagtt tactcatata tactttagat tgatttaaaa cttcattttt
     6181 aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac
     6241 gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag
     6301 atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg
     6361 tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca
     6421 gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac cacttcaaga
     6481 actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca
     6541 gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc
     6601 agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca
     6661 ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa
     6721 aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc
     6781 cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc
     6841 gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg
     6901 cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt cctgcgttat
     6961 cccctgattc tgtggataac cgtattaccg cctttgagtg agctgatacc gctcgccgca
     7021 gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc ggaagagcgc ccaatacgca
     7081 aaccgcctct ccccgcgcgt tggccgattc attaatgcag ctggcacgac aggtttcccg
     7141 actggaaagc gggcagtgag cgcaacgcaa ttaatgtgag ttagctcact cattaggcac
     7201 cccaggcttt acactttatg cttccggctc gtatgttgtg tggaattgtg agcggataac
     7261 aatttcacac aggaaacagc tatgaccatg attacgccaa gctctagcta gaggtcgacc
     7321 aattctcatg tttgacagct tatcatcgca gatccgggca acgttgttgc cattgctgca
     7381 ggcgcagaac tggtaggtat ggaagatcta tacattgaat caatattggc aattagccat
     7441 attagtcatt ggttatatag cataaatcaa tattggctat tggccattgc atacgttgta
     7501 tctatatcat aatatgtaca tttatattgg ctcatgtcca atatgaccgc catgttgaca
     7561 ttgattattg actagttatt aatagtaatc aattacgggg tcattagttc atagcccata
     7621 tatggagttc cgcgttacat aacttacggt aaatggcccg cctggctgac cgcccaacga
     7681 cccccgccca ttgacgtcaa taatgacgta tgttcccata gtaacgccaa tagggacttt
     7741 ccattgacgt caatgggtgg agtatttacg gtaaactgcc cacttggcag tacatcaagt
     7801 gtatcatatg ccaagtccgc cccctattga cgtcaatgac ggtaaatggc ccgcctggca
     7861 ttatgcccag tacatgacct tacgggactt tcctacttgg cagtacatct acgtattagt
     7921 catcgctatt accatggtga tgcggttttg gcagtacacc aatgggcgtg gatagcggtt
     7981 tgactcacgg ggatttccaa gtctccaccc cattgacgtc aatgggagtt tgttttggca
     8041 ccaaaatcaa cgggactttc caaaatgtcg taataacccc gccccgttga cgcaaatggg
     8101 cggtaggcgt gtacggtggg aggtctatat aagcagagct cgtttagtga accgtcagat
     8161 cctcactctc ttccgcatcg ctgtctgcga gggccagctg ttgggctcgc ggttgaggac
     8221 aaactcttcg cggtctttcc agtactcttg gatcggaaac ccgtcggcct ccgaacggta
     8281 ctccgccacc gagggacctg agcgagtccg catcgaccgg atcggaaaac ct
//
