LOCUS       sequence-242839-        7860 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of recombinant P. knowlesi protein in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7860)
  AUTHORS   Muller-Sienerth N, Shilts J, Kadir KA, Yman V, Homann MV, Asghar M, 
            Ngasala B, Singh B, Farnert A, Wright GJ
  TITLE     A panel of recombinant proteins from human-infective Plasmodium 
            species for serological surveillance.
  JOURNAL   Malar J. 2020 Jan 17;19(1):31. doi: 10.1186/s12936-020-3111-5.
  PUBMED    31952523
REFERENCE   2  (bases 1 to 7860)
  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: "Malar J."; 
            date: "2020-01-17"; volume: "19(1):31. doi"; pages: " 
            10.1186/s12936-020-3111-5"
COMMENT     Sequence Label: Pk_MSP10-Cd4-bio-His
FEATURES             Location/Qualifiers
     source          1..7860
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        7..386
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        387..590
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     541..561
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     587..611
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     primer_bind     1092..1111
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     regulatory      1134..1143
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3024..3041
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    3093..3148
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(3147..3166)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(5384..5402)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5470..5574
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5575..5643,5644..6435)
                     /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"
     primer_bind     complement(5793..5812)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6606..7194
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7095..7114
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7348..7365
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    7482..7503
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7518..7523,7524..7541,7542..7548)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7556..7572
                     /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     7561..7583
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7580..7596
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7580..7596
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 catgttgaca ttgattattg actagttatt aatagtaatc aattacgggg tcattagttc
       61 atagcccata tatggagttc cgcgttacat aacttacggt aaatggcccg cctggctgac
      121 cgcccaacga cccccgccca ttgacgtcaa taatgacgta tgttcccata gtaacgccaa
      181 tagggacttt ccattgacgt caatgggtgg agtatttacg gtaaactgcc cacttggcag
      241 tacatcaagt gtatcatatg ccaagtccgc cccctattga cgtcaatgac ggtaaatggc
      301 ccgcctggca ttatgcccag tacatgacct tacgggactt tcctacttgg cagtacatct
      361 acgtattagt catcgctatt accatggtga tgcggttttg gcagtacacc aatgggcgtg
      421 gatagcggtt tgactcacgg ggatttccaa gtctccaccc cattgacgtc aatgggagtt
      481 tgttttggca ccaaaatcaa cgggactttc caaaatgtcg taataacccc gccccgttga
      541 cgcaaatggg cggtaggcgt gtacggtggg aggtctatat aagcagagct cgtttagtga
      601 accgtcagat cctcactctc ttccgcatcg ctgtctgcga gggccagctg ttgggctcgc
      661 ggttgaggac aaactcttcg cggtctttcc agtactcttg gatcggaaac ccgtcggcct
      721 ccgaacggta ctccgccacc gagggacctg agcgagtccg catcgaccgg atcggaaaac
      781 ctctcgagaa aggcgtctaa ccagtcacag tcgcaaggta ggctgagcac cgtggcgggc
      841 ggcagcgggt ggcggtcggg gttgtttctg gcggaggtgc tgctgatgat gtaattaaag
      901 taggcggtct tgagacggcg gatggtcgag gtgaggtgtg gcaggcttga gatccagctg
      961 ttggggtgag tactccctct caaaagcggg cattacttct gcgctaagat tgtcagtttc
     1021 caaaaacgag gaggatttga tattcacctg gcccgatctg gccatacact tgagtgacaa
     1081 tgacatccac tttgcctttc tctccacagg tgtccactcc caggtccaag tttgccacca
     1141 tggagtttca gacccaggta ctcatgtccc tgctgctctg catgtctggt gcggccgcca
     1201 atggcaatga gctgaagggc acagatggcg tggaccctcc caagcaccag gacatcacca
     1261 acgactacaa caccctggaa cacatcagca gcgagggcca gatcagccgg cggaacgacg
     1321 atagcaccat ccctcagagc aacacccagc aggaagagga aggcagcgac gtcaaagagg
     1381 gcaacacctt cctgcacggc aagaacgtgg aaaaagaggt ggaaacctac ctgatccagg
     1441 aagaacagaa caaagtcaac cacgccgacc tgggcaccgg caagcaggaa aaagccagcc
     1501 accacgtgca gaagaacgcc aacctgcgga gcaccagcaa gagcagctct gaggccgccg
     1561 tgaaccaggc ctacaacttc gtgcagcgga acagcgcccc tctgggcaag aacaaggcca
     1621 acatcgaaca ggtggccaga gcccaggacg acatcgacac caccgagaac gacaacaccc
     1681 acgacaccaa agagcacaac gccgacgccg acaacctgaa cggcgagggc gataacgccg
     1741 agaagggcaa taccgagaat gccgacgcca agaaggacaa cgctgaggac gccaacgccg
     1801 aaaaggatat cgccaagaat gtgaacgcag agaaggacat tgccaaaaac gccaatgccg
     1861 ataccggcaa gggcggcgag gaacacgagg aacccagaat cagccctccc gccgagcagc
     1921 ctaccagaaa cgcccctcct ctggaccatg ccgagaagat caagaaccgg ctgctgaaag
     1981 agggccgcga gctgaaagaa accaccagca tgatcgacaa taccgtgtac aacgtcgagc
     2041 agatcatcct caagaccaag ttctacacca ccgccatccg gaacttcgtc ctgttcaaag
     2101 tgaaccacat ctgcgagtac agcaagtgcg gccccaacgc ccggtgctac atcgtggaaa
     2161 aggacaaaga ggaatgccgc tgtatcgcca actacatgcc cgacaacagc gtggactact
     2221 tcaagtgcat ccccaagacc gtgaaggact gcagcaaaga gaacggcaac tgcgacgtga
     2281 acgccgagtg cagcatcgac aagaaagaaa acatcaagtg ccagtgcaac cacggctata
     2341 tcggcgacgg catcttctgc gtgctgggct ctggcgcgcc gtcgacctcc atcacggcct
     2401 ataagagtga gggggagtca gcggagttct ccttcccact caaccttgga gaggaaagcc
     2461 tgcagggaga gttgagatgg aaggcagaga aggctccttc ttcccagtcc tggatcacct
     2521 tctccctaaa gaaccaaaag gtgtctgtgc agaagtctac tagcaacccc aagttccagc
     2581 tgtccgaaac gctcccactc acccttcaga taccccaggt ctcccttcag tttgctggtt
     2641 ctggcaacct gaccctgact ctggacagag ggatactgta tcaggaagtg aacctggtgg
     2701 tgatgaaagt gactcagccc gacagcaaca ctttgacctg tgaggtgatg ggacccacct
     2761 cacccaagat gagactgatc ttgaagcagg agaatcagga ggccagggtc tccaggcagg
     2821 agaaagtgat tcaagtgcag gcccctgaag caggggtgtg gcaatgtcta ctgagtgaag
     2881 gtgaagaggt caagatggac tccaagatcc aggttttatc caaagggttg aattccggca
     2941 gcctgcacca catcctggac gcccagaaga tgctgtggaa ccacagggac aggaacctgc
     3001 ctcctctggc ccctctgggc ccccaccacc atcaccacca ttgagtctag acacgtgatt
     3061 aattaaggat cccccgacct cgacctctgg ctaataaagg aaatttattt tcattgcaat
     3121 agtgtgttgg aattttttgt gtctctcact cggaaggaca tatgggaggg caaatcattt
     3181 ggtcgagatc cctcggagat ctctagctag aggatcgatc cccgccccgg acgaactaaa
     3241 cctgactacg acatctctgc cccttcttcg cggggcagtg catgtaatcc cttcagttgg
     3301 ttggtacaac ttgccaactg ggccctgttc cacatgtgac acgggggggg accaaacaca
     3361 aaggggttct ctgactgtag ttgacatcct tataaatgga tgtgcacatt tgccaacact
     3421 gagtggcttt catcctggag cagactttgc agtctgtgga ctgcaacaca acattgcctt
     3481 tatgtgtaac tcttggctga agctcttaca ccaatgctgg gggacatgta cctcccaggg
     3541 gcccaggaag actacgggag gctacaccaa cgtcaatcag aggggcctgt gtagctaccg
     3601 ataagcggac cctcaagagg gcattagcaa tagtgtttat aaggccccct tgttaaccct
     3661 aaacgggtag catatgcttc ccgggtagta gtatatacta tccagactaa ccctaattca
     3721 atagcatatg ttacccaacg ggaagcatat gctatcgaat tagggttagt aaaagggtcc
     3781 taaggaacag cgatatctcc caccccatga gctgtcacgg ttttatttac atggggtcag
     3841 gattccacga gggtagtgaa ccattttagt cacaagggca gtggctgaag atcaaggagc
     3901 gggcagtgaa ctctcctgaa tcttcgcctg cttcttcatt ctccttcgtt tagctaatag
     3961 aataactgct gagttgtgaa cagtaaggtg tatgtgaggt gctcgaaaac aaggtttcag
     4021 gtgacgcccc cagaataaaa tttggacggg gggttcagtg gtggcattgt gctatgacac
     4081 caatataacc ctcacaaacc ccttgggcaa taaatactag tgtaggaatg aaacattctg
     4141 aatatcttta acaatagaaa tccatggggt ggggacaagc cgtaaagact ggatgtccat
     4201 ctcacacgaa tttatggcta tgggcaacac ataatcctag tgcaatatga tactggggtt
     4261 attaagatgt gtcccaggca gggaccaaga caggtgaacc atgttgttac actctatttg
     4321 taacaagggg aaagagagtg gacgccgaca gcagcggact ccactggttg tctctaacac
     4381 ccccgaaaat taaacggggc tccacgccaa tggggcccat aaacaaagac aagtggccac
     4441 tctttttttt gaaattgtgg agtgggggca cgcgtcagcc cccacacgcc gccctgcggt
     4501 tttggactgt aaaataaggg tgtaataact tggctgattg taaccccgct aaccactgcg
     4561 gtcaaaccac ttgcccacaa aaccactaat ggcaccccgg ggaatacctg cataagtagg
     4621 tgggcgggcc aagatagggg cgcgattgct gcgatctgga ggacaaatta cacacacttg
     4681 cgcctgagcg ccaagcacag ggttgttggt cctcatattc acgaggtcgc tgagagcacg
     4741 gtgggctaat gttgccatgg gtagcatata ctacccaaat atctggatag catatgctat
     4801 cctaatctat atctgggtag cataggctat cctaatctat atctgggtag catatgctat
     4861 cctaatctat atctgggtag tatatgctat cctaatttat atctgggtag cataggctat
     4921 cctaatctat atctgggtag catatgctat cctaatctat atctgggtag tatatgctat
     4981 cctaatctgt atccgggtag catatgctat cctaatagag attagggtag tatatgctat
     5041 cctaatttat atctgggtag catatactac ccaaatatct ggatagcata tgctatccta
     5101 atctatatct gggtagcata tgctatccta atctatatct gggtagcata ggctatccta
     5161 atctatatct gggtagcata tgctatccta atctatatct gggtagtata tgctatccta
     5221 atttatatct gggtagcata ggctatccta atctatatct gggtagcata tgctatccta
     5281 atctatatct gggtagtata tgctatccta atctgtatcc gggtagcata tgctatcctc
     5341 atgataagct gtcaaacatg agaattttct tgaagacgaa agggcctcgt gatacgccta
     5401 tttttatagg ttaatgtcat gataataatg gtttcttaga cgtcaggtgg cacttttcgg
     5461 ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg
     5521 ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtatgagt
     5581 attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt
     5641 gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg
     5701 ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa
     5761 cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtgtt
     5821 gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag
     5881 tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt
     5941 gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac gatcggagga
     6001 ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt
     6061 tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgca
     6121 gcaatggcaa caacgttgcg caaactatta actggcgaac tacttactct agcttcccgg
     6181 caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc
     6241 cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt
     6301 atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg
     6361 gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg
     6421 attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat tgatttaaaa
     6481 cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa
     6541 atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga
     6601 tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg
     6661 ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact
     6721 ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac
     6781 cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg
     6841 gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg
     6901 gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga
     6961 acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc
     7021 gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg
     7081 agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc
     7141 tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc
     7201 agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt
     7261 cctgcgttat cccctgattc tgtggataac cgtattaccg cctttgagtg agctgatacc
     7321 gctcgccgca gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc ggaagagcgc
     7381 ccaatacgca aaccgcctct ccccgcgcgt tggccgattc attaatgcag ctggcacgac
     7441 aggtttcccg actggaaagc gggcagtgag cgcaacgcaa ttaatgtgag ttagctcact
     7501 cattaggcac cccaggcttt acactttatg cttccggctc gtatgttgtg tggaattgtg
     7561 agcggataac aatttcacac aggaaacagc tatgaccatg attacgccaa gctctagcta
     7621 gaggtcgacc aattctcatg tttgacagct tatcatcgca gatccgggca acgttgttgc
     7681 cattgctgca ggcgcagaac tggtaggtat ggaagatcta tacattgaat caatattggc
     7741 aattagccat attagtcatt ggttatatag cataaatcaa tattggctat tggccattgc
     7801 atacgttgta tctatatcat aatatgtaca tttatattgg ctcatgtcca atatgaccgc
//
