LOCUS       sequence-208812-        9655 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 9655)
  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 9655)
  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: PF3D7_1106200-COMP-blac-flag-his
FEATURES             Location/Qualifiers
     source          1..9655
                     /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             3187..3972
                     /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             3979..4044
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             4051..4068
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    4106..4161
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(4160..4179)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(6397..6415)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6483..6587
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6588..6656,6657..7448)
                     /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      7619..8207
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8108..8127
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     8361..8378
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    8495..8516
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(8531..8536,8537..8554,8555..8561)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8569..8585
                     /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     8574..8596
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     8593..8609
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     8593..8609
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     enhancer        8880..9259
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        9260..9463
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     9414..9434
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     9460..9484
                     /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 ggccgccaac
      421 aaagaaatcg ccatcctgtg cagcaaagtg cacctgaaga agcacatcta ctaccccaga
      481 aagggcagcc acttccacat ctacagcgtg gtggacaaca acgtgaacct caagaacaag
      541 aacgagtaca acagcaagct gaaggaccgg atcaagatct acgaacaggt gcacaacaac
      601 aagtgcaaga agatctactt cctgcccctg atcttcctga agaacaacaa taaggacgtg
      661 tacatcgagg accacaagga caagaacatc gataacaata ataacaacaa gaaagataac
      721 aagaaggacg acatcaacaa caacgacaag aagaacaaaa tcaacgacgc ccctgcccag
      781 aatatcgcca acgataagga taagaactac gccaacaact tcgagtatat cgagaacaac
      841 ctgctgaagc tgaaaaacaa cctggaagtg ttcagcagct atatcaacaa ggccgacgtg
      901 atcatcgtgc ccctgaacta ccacgacatc ctgaacaagc acctgatcga gatcatccct
      961 ttcaacaaca tcatcaacag cctggactac atcttcctgt tcaacaagaa tgactataac
     1021 aaagccgaca tcaatattaa caacaacaac aacaataaca acaacatgaa caagtacaac
     1081 atctgccctc cctacaacaa tatctacaat atcatcttct tcatctacaa ctacaacgac
     1141 caggacaacc ctctgaacat tgccagctac acccctagct ttttcaccaa caaaaaggac
     1201 aacattatcc acaaagagaa gttccagctg gtccacgtga tccagaagtt tctgaacgac
     1261 cactacaacc tcaaggatca gaatgagcac aagaagtacc tgaacatcaa ctttctgttc
     1321 accaaggaca atatcctgga cattaacaag atcgacaagg atatcgaaga gaacaagatg
     1381 ctgtactacc ctaagaagaa tatcctcaac aaagagacaa actacgtcaa cacagccaac
     1441 aacgctaaca atgccaacaa gcaggaagat cagaacctga tcaaatacgt ccacaagaac
     1501 aatgacgcct tctttaagaa gtacaagccc gacaccctga agaatctggc caacgagaac
     1561 tactttatct ataacttcct gagcagctac aagctggaca tcctcaaaga gtacatctac
     1621 attgccaagg acatcaccat cccaaacaac aatgtgatca agcacatcca gaacattaac
     1681 atcaagaaga agatccacaa tatcgagctg ctgctcaaca atttcaagaa ctgcaagctc
     1741 aacaaaaaca tcaatagcca cgtgaacgtg caggaaatca agaaaaagca gtacagctac
     1801 ctgctgcaca tcatcctgag cgacctgctc gagaagtacg acagcaatct ggaaattctg
     1861 aagaactacc tgtaccaggt gttcaaagag aatatccaga agatcaagat taactccgat
     1921 atcctgttcc agtttaagaa gcagatcaac cacatcgacc agctgttcga ctactacaac
     1981 tccaagatct tcatcctccc cagcaacaag aactatcagg cccagcagaa gtcctactac
     2041 atctattact actataagat caagctgctg cagctcctga accaggccgc caatgagatc
     2101 atcaattact acatcagcaa gggcctgtac gtgaagaatt acgccttcaa cgagttcagc
     2161 ttctccaaga agtataccct gatcaattac atcatcacct acatctttaa cctgattaag
     2221 aacaaagtga atatcaactt caactacctg agccccaacg ccttcggctt cagctcctac
     2281 aaggacgata tcagcctgag ccctaagaaa gacatcatga tcaccaccag cgagatggac
     2341 caggtccaga aaatcaacat ccccagaagc gcctacacca agaccatgct gatgatggac
     2401 aaggacagaa acggcgcgcc gtcgacctcc atcacggcct ataagagtga gggggagtca
     2461 gcggagttct ccttcccact caaccttgga gaggaaagcc tgcagggaga gttgagatgg
     2521 aaggcagaga aggctccttc ttcccagtcc tggatcacct tctccctaaa gaaccaaaag
     2581 gtgtctgtgc agaagtctac tagcaacccc aagttccagc tgtccgaaac gctcccactc
     2641 acccttcaga taccccaggt ctcccttcag tttgctggtt ctggcaacct gaccctgact
     2701 ctggacagag ggatactgta tcaggaagtg aacctggtgg tgatgaaagt gactcagccc
     2761 gacagcaaca ctttgacctg tgaggtgatg ggacccacct cacccaagat gagactgatc
     2821 ttgaagcagg agaatcagga ggccagggtc tccaggcagg agaaagtgat tcaagtgcag
     2881 gcccctgaag caggggtgtg gcaatgtcta ctgagtgaag gtgaagaggt caagatggac
     2941 tccaagatcc aggttttatc caaagggttg aattccggcg gcggctccgg aggcggtacc
     3001 ggtggcggcg acctagcccc acagatgctt cgagaactcc aggagactaa tgcggcgctg
     3061 caagacgtga gagagctctt gcgacagcag gtcaaggaga tcaccttcct gaagaatacg
     3121 gtgatggaat gtgacgcctg cggtctagac agaaacctgc ctccactagc acctctgggc
     3181 ccacacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg
     3241 ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa
     3301 cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt
     3361 gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag
     3421 tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt
     3481 gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac gatcggagga
     3541 ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt
     3601 tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgta
     3661 gcaatggcaa caacgttgcg caaactatta actggcgaac tacttactct agcttcccgg
     3721 caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc
     3781 cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt
     3841 atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg
     3901 gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg
     3961 attaagcatt ggggccctga ctacaaagac catgacggtg attataaaga tcatgacatc
     4021 gattacaagg atgacgatga caagggccct caccaccacc atcaccacta attaattaag
     4081 gatcccccga cctcgacctc tggctaataa aggaaattta ttttcattgc aatagtgtgt
     4141 tggaattttt tgtgtctctc actcggaagg acatatggga gggcaaatca tttggtcgag
     4201 atccctcgga gatctctagc tagaggatcg atccccgccc cggacgaact aaacctgact
     4261 acgacatctc tgccccttct tcgcggggca gtgcatgtaa tcccttcagt tggttggtac
     4321 aacttgccaa ctgggccctg ttccacatgt gacacggggg gggaccaaac acaaaggggt
     4381 tctctgactg tagttgacat ccttataaat ggatgtgcac atttgccaac actgagtggc
     4441 tttcatcctg gagcagactt tgcagtctgt ggactgcaac acaacattgc ctttatgtgt
     4501 aactcttggc tgaagctctt acaccaatgc tgggggacat gtacctccca ggggcccagg
     4561 aagactacgg gaggctacac caacgtcaat cagaggggcc tgtgtagcta ccgataagcg
     4621 gaccctcaag agggcattag caatagtgtt tataaggccc ccttgttaac cctaaacggg
     4681 tagcatatgc ttcccgggta gtagtatata ctatccagac taaccctaat tcaatagcat
     4741 atgttaccca acgggaagca tatgctatcg aattagggtt agtaaaaggg tcctaaggaa
     4801 cagcgatatc tcccacccca tgagctgtca cggttttatt tacatggggt caggattcca
     4861 cgagggtagt gaaccatttt agtcacaagg gcagtggctg aagatcaagg agcgggcagt
     4921 gaactctcct gaatcttcgc ctgcttcttc attctccttc gtttagctaa tagaataact
     4981 gctgagttgt gaacagtaag gtgtatgtga ggtgctcgaa aacaaggttt caggtgacgc
     5041 ccccagaata aaatttggac ggggggttca gtggtggcat tgtgctatga caccaatata
     5101 accctcacaa accccttggg caataaatac tagtgtagga atgaaacatt ctgaatatct
     5161 ttaacaatag aaatccatgg ggtggggaca agccgtaaag actggatgtc catctcacac
     5221 gaatttatgg ctatgggcaa cacataatcc tagtgcaata tgatactggg gttattaaga
     5281 tgtgtcccag gcagggacca agacaggtga accatgttgt tacactctat ttgtaacaag
     5341 gggaaagaga gtggacgccg acagcagcgg actccactgg ttgtctctaa cacccccgaa
     5401 aattaaacgg ggctccacgc caatggggcc cataaacaaa gacaagtggc cactcttttt
     5461 tttgaaattg tggagtgggg gcacgcgtca gcccccacac gccgccctgc ggttttggac
     5521 tgtaaaataa gggtgtaata acttggctga ttgtaacccc gctaaccact gcggtcaaac
     5581 cacttgccca caaaaccact aatggcaccc cggggaatac ctgcataagt aggtgggcgg
     5641 gccaagatag gggcgcgatt gctgcgatct ggaggacaaa ttacacacac ttgcgcctga
     5701 gcgccaagca cagggttgtt ggtcctcata ttcacgaggt cgctgagagc acggtgggct
     5761 aatgttgcca tgggtagcat atactaccca aatatctgga tagcatatgc tatcctaatc
     5821 tatatctggg tagcataggc tatcctaatc tatatctggg tagcatatgc tatcctaatc
     5881 tatatctggg tagtatatgc tatcctaatt tatatctggg tagcataggc tatcctaatc
     5941 tatatctggg tagcatatgc tatcctaatc tatatctggg tagtatatgc tatcctaatc
     6001 tgtatccggg tagcatatgc tatcctaata gagattaggg tagtatatgc tatcctaatt
     6061 tatatctggg tagcatatac tacccaaata tctggatagc atatgctatc ctaatctata
     6121 tctgggtagc atatgctatc ctaatctata tctgggtagc ataggctatc ctaatctata
     6181 tctgggtagc atatgctatc ctaatctata tctgggtagt atatgctatc ctaatttata
     6241 tctgggtagc ataggctatc ctaatctata tctgggtagc atatgctatc ctaatctata
     6301 tctgggtagt atatgctatc ctaatctgta tccgggtagc atatgctatc ctcatgataa
     6361 gctgtcaaac atgagaattt tcttgaagac gaaagggcct cgtgatacgc ctatttttat
     6421 aggttaatgt catgataata atggtttctt agacgtcagg tggcactttt cggggaaatg
     6481 tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga
     6541 gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtatg agtattcaac
     6601 atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt tttgctcacc
     6661 cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcacga gtgggttaca
     6721 tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc
     6781 caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt gttgacgccg
     6841 ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt gagtactcac
     6901 cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc agtgctgcca
     6961 taaccatgag tgataacact gcggccaact tacttctgac aacgatcgga ggaccgaagg
     7021 agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat cgttgggaac
     7081 cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct gcagcaatgg
     7141 caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc cggcaacaat
     7201 taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg gcccttccgg
     7261 ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc ggtatcattg
     7321 cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg acggggagtc
     7381 aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc
     7441 attggtaact gtcagaccaa gtttactcat atatacttta gattgattta aaacttcatt
     7501 tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt
     7561 aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt
     7621 gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag
     7681 cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta actggcttca
     7741 gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca
     7801 agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg
     7861 ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg
     7921 cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct
     7981 acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga
     8041 gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc
     8101 ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg
     8161 agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg
     8221 cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc tttcctgcgt
     8281 tatcccctga ttctgtggat aaccgtatta ccgcctttga gtgagctgat accgctcgcc
     8341 gcagccgaac gaccgagcgc agcgagtcag tgagcgagga agcggaagag cgcccaatac
     8401 gcaaaccgcc tctccccgcg cgttggccga ttcattaatg cagctggcac gacaggtttc
     8461 ccgactggaa agcgggcagt gagcgcaacg caattaatgt gagttagctc actcattagg
     8521 caccccaggc tttacacttt atgcttccgg ctcgtatgtt gtgtggaatt gtgagcggat
     8581 aacaatttca cacaggaaac agctatgacc atgattacgc caagctctag ctagaggtcg
     8641 accaattctc atgtttgaca gcttatcatc gcagatccgg gcaacgttgt tgccattgct
     8701 gcaggcgcag aactggtagg tatggaagat ctatacattg aatcaatatt ggcaattagc
     8761 catattagtc attggttata tagcataaat caatattggc tattggccat tgcatacgtt
     8821 gtatctatat cataatatgt acatttatat tggctcatgt ccaatatgac cgccatgttg
     8881 acattgatta ttgactagtt attaatagta atcaattacg gggtcattag ttcatagccc
     8941 atatatggag ttccgcgtta cataacttac ggtaaatggc ccgcctggct gaccgcccaa
     9001 cgacccccgc ccattgacgt caataatgac gtatgttccc atagtaacgc caatagggac
     9061 tttccattga cgtcaatggg tggagtattt acggtaaact gcccacttgg cagtacatca
     9121 agtgtatcat atgccaagtc cgccccctat tgacgtcaat gacggtaaat ggcccgcctg
     9181 gcattatgcc cagtacatga ccttacggga ctttcctact tggcagtaca tctacgtatt
     9241 agtcatcgct attaccatgg tgatgcggtt ttggcagtac accaatgggc gtggatagcg
     9301 gtttgactca cggggatttc caagtctcca ccccattgac gtcaatggga gtttgttttg
     9361 gcaccaaaat caacgggact ttccaaaatg tcgtaataac cccgccccgt tgacgcaaat
     9421 gggcggtagg cgtgtacggt gggaggtcta tataagcaga gctcgtttag tgaaccgtca
     9481 gatcctcact ctcttccgca tcgctgtctg cgagggccag ctgttgggct cgcggttgag
     9541 gacaaactct tcgcggtctt tccagtactc ttggatcgga aacccgtcgg cctccgaacg
     9601 gtactccgcc accgagggac ctgagcgagt ccgcatcgac cggatcggaa aacct
//
