LOCUS       sequence-208689-        8005 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 8005)
  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 8005)
  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: TRSP-COMP-blac-flag-his
FEATURES             Location/Qualifiers
     source          1..8005
                     /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             1537..2322
                     /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             2329..2394
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             2401..2418
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    2456..2511
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(2510..2529)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(4747..4765)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4833..4937
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4938..5006,5007..5798)
                     /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      5969..6557
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6458..6477
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6711..6728
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    6845..6866
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6881..6886,6887..6904,6905..6911)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6919..6935
                     /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     6924..6946
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     6943..6959
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6943..6959
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     enhancer        7230..7609
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        7610..7813
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     7764..7784
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     7810..7834
                     /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 ggccgccgac
      421 ttcttcctgc ggtatcggac cggcttcatc agaagccggc tggaaaccta catcggcaac
      481 agcgacgtgc ggtacaacaa ggccttcatc aacaaccggc tgctgaacga gcacgcccac
      541 tgcgacgctt ggagcgagtg gtccgcctgc agcaagacct gcgactacgg catcaagatc
      601 agagtgaaga tcagcaccga ccagaccaag agcaaggcct gctccaatat cgccgagagc
      661 accatctgcc acgagcacat ctgtccccgg accttcgagg aagccgagga aacatacctg
      721 cacaacaaag agaaagagaa gaagaaaaag ttccggacca ccggcgcgcc gtcgacctcc
      781 atcacggcct ataagagtga gggggagtca gcggagttct ccttcccact caaccttgga
      841 gaggaaagcc tgcagggaga gttgagatgg aaggcagaga aggctccttc ttcccagtcc
      901 tggatcacct tctccctaaa gaaccaaaag gtgtctgtgc agaagtctac tagcaacccc
      961 aagttccagc tgtccgaaac gctcccactc acccttcaga taccccaggt ctcccttcag
     1021 tttgctggtt ctggcaacct gaccctgact ctggacagag ggatactgta tcaggaagtg
     1081 aacctggtgg tgatgaaagt gactcagccc gacagcaaca ctttgacctg tgaggtgatg
     1141 ggacccacct cacccaagat gagactgatc ttgaagcagg agaatcagga ggccagggtc
     1201 tccaggcagg agaaagtgat tcaagtgcag gcccctgaag caggggtgtg gcaatgtcta
     1261 ctgagtgaag gtgaagaggt caagatggac tccaagatcc aggttttatc caaagggttg
     1321 aattccggcg gcggctccgg aggcggtacc ggtggcggcg acctagcccc acagatgctt
     1381 cgagaactcc aggagactaa tgcggcgctg caagacgtga gagagctctt gcgacagcag
     1441 gtcaaggaga tcaccttcct gaagaatacg gtgatggaat gtgacgcctg cggtctagac
     1501 agaaacctgc ctccactagc acctctgggc ccacacccag aaacgctggt gaaagtaaaa
     1561 gatgctgaag atcagttggg tgcacgagtg ggttacatcg aactggatct caacagcggt
     1621 aagatccttg agagttttcg ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt
     1681 ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc aagagcaact cggtcgccgc
     1741 atacactatt ctcagaatga cttggttgag tactcaccag tcacagaaaa gcatcttacg
     1801 gatggcatga cagtaagaga attatgcagt gctgccataa ccatgagtga taacactgcg
     1861 gccaacttac ttctgacaac gatcggagga ccgaaggagc taaccgcttt tttgcacaac
     1921 atgggggatc atgtaactcg ccttgatcgt tgggaaccgg agctgaatga agccatacca
     1981 aacgacgagc gtgacaccac gatgcctgta gcaatggcaa caacgttgcg caaactatta
     2041 actggcgaac tacttactct agcttcccgg caacaattaa tagactggat ggaggcggat
     2101 aaagttgcag gaccacttct gcgctcggcc cttccggctg gctggtttat tgctgataaa
     2161 tctggagccg gtgagcgtgg gtctcgcggt atcattgcag cactggggcc agatggtaag
     2221 ccctcccgta tcgtagttat ctacacgacg gggagtcagg caactatgga tgaacgaaat
     2281 agacagatcg ctgagatagg tgcctcactg attaagcatt ggggccctga ctacaaagac
     2341 catgacggtg attataaaga tcatgacatc gattacaagg atgacgatga caagggccct
     2401 caccaccacc atcaccacta attaattaag gatcccccga cctcgacctc tggctaataa
     2461 aggaaattta ttttcattgc aatagtgtgt tggaattttt tgtgtctctc actcggaagg
     2521 acatatggga gggcaaatca tttggtcgag atccctcgga gatctctagc tagaggatcg
     2581 atccccgccc cggacgaact aaacctgact acgacatctc tgccccttct tcgcggggca
     2641 gtgcatgtaa tcccttcagt tggttggtac aacttgccaa ctgggccctg ttccacatgt
     2701 gacacggggg gggaccaaac acaaaggggt tctctgactg tagttgacat ccttataaat
     2761 ggatgtgcac atttgccaac actgagtggc tttcatcctg gagcagactt tgcagtctgt
     2821 ggactgcaac acaacattgc ctttatgtgt aactcttggc tgaagctctt acaccaatgc
     2881 tgggggacat gtacctccca ggggcccagg aagactacgg gaggctacac caacgtcaat
     2941 cagaggggcc tgtgtagcta ccgataagcg gaccctcaag agggcattag caatagtgtt
     3001 tataaggccc ccttgttaac cctaaacggg tagcatatgc ttcccgggta gtagtatata
     3061 ctatccagac taaccctaat tcaatagcat atgttaccca acgggaagca tatgctatcg
     3121 aattagggtt agtaaaaggg tcctaaggaa cagcgatatc tcccacccca tgagctgtca
     3181 cggttttatt tacatggggt caggattcca cgagggtagt gaaccatttt agtcacaagg
     3241 gcagtggctg aagatcaagg agcgggcagt gaactctcct gaatcttcgc ctgcttcttc
     3301 attctccttc gtttagctaa tagaataact gctgagttgt gaacagtaag gtgtatgtga
     3361 ggtgctcgaa aacaaggttt caggtgacgc ccccagaata aaatttggac ggggggttca
     3421 gtggtggcat tgtgctatga caccaatata accctcacaa accccttggg caataaatac
     3481 tagtgtagga atgaaacatt ctgaatatct ttaacaatag aaatccatgg ggtggggaca
     3541 agccgtaaag actggatgtc catctcacac gaatttatgg ctatgggcaa cacataatcc
     3601 tagtgcaata tgatactggg gttattaaga tgtgtcccag gcagggacca agacaggtga
     3661 accatgttgt tacactctat ttgtaacaag gggaaagaga gtggacgccg acagcagcgg
     3721 actccactgg ttgtctctaa cacccccgaa aattaaacgg ggctccacgc caatggggcc
     3781 cataaacaaa gacaagtggc cactcttttt tttgaaattg tggagtgggg gcacgcgtca
     3841 gcccccacac gccgccctgc ggttttggac tgtaaaataa gggtgtaata acttggctga
     3901 ttgtaacccc gctaaccact gcggtcaaac cacttgccca caaaaccact aatggcaccc
     3961 cggggaatac ctgcataagt aggtgggcgg gccaagatag gggcgcgatt gctgcgatct
     4021 ggaggacaaa ttacacacac ttgcgcctga gcgccaagca cagggttgtt ggtcctcata
     4081 ttcacgaggt cgctgagagc acggtgggct aatgttgcca tgggtagcat atactaccca
     4141 aatatctgga tagcatatgc tatcctaatc tatatctggg tagcataggc tatcctaatc
     4201 tatatctggg tagcatatgc tatcctaatc tatatctggg tagtatatgc tatcctaatt
     4261 tatatctggg tagcataggc tatcctaatc tatatctggg tagcatatgc tatcctaatc
     4321 tatatctggg tagtatatgc tatcctaatc tgtatccggg tagcatatgc tatcctaata
     4381 gagattaggg tagtatatgc tatcctaatt tatatctggg tagcatatac tacccaaata
     4441 tctggatagc atatgctatc ctaatctata tctgggtagc atatgctatc ctaatctata
     4501 tctgggtagc ataggctatc ctaatctata tctgggtagc atatgctatc ctaatctata
     4561 tctgggtagt atatgctatc ctaatttata tctgggtagc ataggctatc ctaatctata
     4621 tctgggtagc atatgctatc ctaatctata tctgggtagt atatgctatc ctaatctgta
     4681 tccgggtagc atatgctatc ctcatgataa gctgtcaaac atgagaattt tcttgaagac
     4741 gaaagggcct cgtgatacgc ctatttttat aggttaatgt catgataata atggtttctt
     4801 agacgtcagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct
     4861 aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat
     4921 attgaaaaag gaagagtatg agtattcaac atttccgtgt cgcccttatt cccttttttg
     4981 cggcattttg ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta aaagatgctg
     5041 aagatcagtt gggtgcacga gtgggttaca tcgaactgga tctcaacagc ggtaagatcc
     5101 ttgagagttt tcgccccgaa gaacgttttc caatgatgag cacttttaaa gttctgctat
     5161 gtggcgcggt attatcccgt gttgacgccg ggcaagagca actcggtcgc cgcatacact
     5221 attctcagaa tgacttggtt gagtactcac cagtcacaga aaagcatctt acggatggca
     5281 tgacagtaag agaattatgc agtgctgcca taaccatgag tgataacact gcggccaact
     5341 tacttctgac aacgatcgga ggaccgaagg agctaaccgc ttttttgcac aacatggggg
     5401 atcatgtaac tcgccttgat cgttgggaac cggagctgaa tgaagccata ccaaacgacg
     5461 agcgtgacac cacgatgcct gcagcaatgg caacaacgtt gcgcaaacta ttaactggcg
     5521 aactacttac tctagcttcc cggcaacaat taatagactg gatggaggcg gataaagttg
     5581 caggaccact tctgcgctcg gcccttccgg ctggctggtt tattgctgat aaatctggag
     5641 ccggtgagcg tgggtctcgc ggtatcattg cagcactggg gccagatggt aagccctccc
     5701 gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga
     5761 tcgctgagat aggtgcctca ctgattaagc attggtaact gtcagaccaa gtttactcat
     5821 atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc
     5881 tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag
     5941 accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct
     6001 gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac
     6061 caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc
     6121 tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg
     6181 ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt
     6241 tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt
     6301 gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc
     6361 tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca
     6421 gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata
     6481 gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg
     6541 ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct
     6601 ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta
     6661 ccgcctttga gtgagctgat accgctcgcc gcagccgaac gaccgagcgc agcgagtcag
     6721 tgagcgagga agcggaagag cgcccaatac gcaaaccgcc tctccccgcg cgttggccga
     6781 ttcattaatg cagctggcac gacaggtttc ccgactggaa agcgggcagt gagcgcaacg
     6841 caattaatgt gagttagctc actcattagg caccccaggc tttacacttt atgcttccgg
     6901 ctcgtatgtt gtgtggaatt gtgagcggat aacaatttca cacaggaaac agctatgacc
     6961 atgattacgc caagctctag ctagaggtcg accaattctc atgtttgaca gcttatcatc
     7021 gcagatccgg gcaacgttgt tgccattgct gcaggcgcag aactggtagg tatggaagat
     7081 ctatacattg aatcaatatt ggcaattagc catattagtc attggttata tagcataaat
     7141 caatattggc tattggccat tgcatacgtt gtatctatat cataatatgt acatttatat
     7201 tggctcatgt ccaatatgac cgccatgttg acattgatta ttgactagtt attaatagta
     7261 atcaattacg gggtcattag ttcatagccc atatatggag ttccgcgtta cataacttac
     7321 ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc ccattgacgt caataatgac
     7381 gtatgttccc atagtaacgc caatagggac tttccattga cgtcaatggg tggagtattt
     7441 acggtaaact gcccacttgg cagtacatca agtgtatcat atgccaagtc cgccccctat
     7501 tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc cagtacatga ccttacggga
     7561 ctttcctact tggcagtaca tctacgtatt agtcatcgct attaccatgg tgatgcggtt
     7621 ttggcagtac accaatgggc gtggatagcg gtttgactca cggggatttc caagtctcca
     7681 ccccattgac gtcaatggga gtttgttttg gcaccaaaat caacgggact ttccaaaatg
     7741 tcgtaataac cccgccccgt tgacgcaaat gggcggtagg cgtgtacggt gggaggtcta
     7801 tataagcaga gctcgtttag tgaaccgtca gatcctcact ctcttccgca tcgctgtctg
     7861 cgagggccag ctgttgggct cgcggttgag gacaaactct tcgcggtctt tccagtactc
     7921 ttggatcgga aacccgtcgg cctccgaacg gtactccgcc accgagggac ctgagcgagt
     7981 ccgcatcgac cggatcggaa aacct
//
