LOCUS       sequence-245944-        7686 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 7686)
  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 7686)
  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_MSP5-Cd4-bio-His
FEATURES             Location/Qualifiers
     source          1..7686
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..380
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        381..584
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     535..555
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     581..605
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     primer_bind     1086..1105
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     regulatory      1128..1137
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2844..2861
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    2913..2968
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(2967..2986)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(5204..5222)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5290..5394
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5395..5463,5464..6255)
                     /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(5613..5632)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6426..7014
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6915..6934
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7168..7185
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    7302..7323
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7338..7343,7344..7361,7362..7368)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7376..7392
                     /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     7381..7403
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7400..7416
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7400..7416
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc
       61 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
      121 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
      181 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
      241 aagtgtatca tatgccaagt ccgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
      301 ggcattatgc ccagtacatg accttacggg actttcctac ttggcagtac atctacgtat
      361 tagtcatcgc tattaccatg gtgatgcggt tttggcagta caccaatggg cgtggatagc
      421 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt
      481 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaataa ccccgccccg ttgacgcaaa
      541 tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctcgttta gtgaaccgtc
      601 agatcctcac tctcttccgc atcgctgtct gcgagggcca gctgttgggc tcgcggttga
      661 ggacaaactc ttcgcggtct ttccagtact cttggatcgg aaacccgtcg gcctccgaac
      721 ggtactccgc caccgaggga cctgagcgag tccgcatcga ccggatcgga aaacctctcg
      781 agaaaggcgt ctaaccagtc acagtcgcaa ggtaggctga gcaccgtggc gggcggcagc
      841 gggtggcggt cggggttgtt tctggcggag gtgctgctga tgatgtaatt aaagtaggcg
      901 gtcttgagac ggcggatggt cgaggtgagg tgtggcaggc ttgagatcca gctgttgggg
      961 tgagtactcc ctctcaaaag cgggcattac ttctgcgcta agattgtcag tttccaaaaa
     1021 cgaggaggat ttgatattca cctggcccga tctggccata cacttgagtg acaatgacat
     1081 ccactttgcc tttctctcca caggtgtcca ctcccaggtc caagtttgcc accatggagt
     1141 ttcagaccca ggtactcatg tccctgctgc tctgcatgtc tggtgcggcc gcccatcctc
     1201 tgaagttcag cttctggcgg gaagagaaga tccacctcca gatctacacc aaccggctgc
     1261 tgagagagga aggcaagaac gacaacgtgg aacagatgga aagccccagc atcagcggca
     1321 ccgagggcaa gaaagaaatc cagatgatca gccatctgca gctccagagc ggcaagcacg
     1381 accaggtgtc ctttctgagc gccaacgacg ccaacctgaa gggcgccaat gaggaagcct
     1441 ctgccgagag cggcgagaac ggcaagaagt ccgacgagga aaacgtgaag aaaagcgacg
     1501 aagagaacgc caaaaagtct gatgaagaga acaaggacgc caacagcaac accaaggatg
     1561 ccgagtccgc cgagggcgaa gagaatcccg tgtcccagga aaaccagatg aagaccctga
     1621 acaacgaaga ggacaagacc aacgatggca agaagaacgg ggacgaggaa gataagaagg
     1681 gcgaaaaggg caaggacgac ggcaagggcg aggaaggctc cgaggaaaag atggaagagt
     1741 ccgccaagaa agaggaagag aataccgagg ccaacaagaa cctggaaagc aagcacgccg
     1801 aggccgaggt gtccgagcac aagcaggatg tgacaacccc tgagggcggc aaggatagcc
     1861 ccgaaggcga gaaccagaaa gagaacaccc ctaacaacgc caaccaggaa aatcaggaaa
     1921 acgccgacaa cgacgagaac aagctgcacc tggacaagct ggacgacgac gtgccacact
     1981 acagcgccct gcggaacaac cgggtggaca agggcatcac cgacagcatg atgctgaaga
     2041 acatcatcga ggataacacc aagagctgca gcgtgaacaa cggcggctgc gccgacgacc
     2101 agatctgcat ccggatcaac aacatgggca tcaagtgcat ctgcaaagag ggccacctgt
     2161 tcggcggcaa gtgtatcctg agcagaagct ctggcgcgcc gtcgacctcc atcacggcct
     2221 ataagagtga gggggagtca gcggagttct ccttcccact caaccttgga gaggaaagcc
     2281 tgcagggaga gttgagatgg aaggcagaga aggctccttc ttcccagtcc tggatcacct
     2341 tctccctaaa gaaccaaaag gtgtctgtgc agaagtctac tagcaacccc aagttccagc
     2401 tgtccgaaac gctcccactc acccttcaga taccccaggt ctcccttcag tttgctggtt
     2461 ctggcaacct gaccctgact ctggacagag ggatactgta tcaggaagtg aacctggtgg
     2521 tgatgaaagt gactcagccc gacagcaaca ctttgacctg tgaggtgatg ggacccacct
     2581 cacccaagat gagactgatc ttgaagcagg agaatcagga ggccagggtc tccaggcagg
     2641 agaaagtgat tcaagtgcag gcccctgaag caggggtgtg gcaatgtcta ctgagtgaag
     2701 gtgaagaggt caagatggac tccaagatcc aggttttatc caaagggttg aattccggca
     2761 gcctgcacca catcctggac gcccagaaga tgctgtggaa ccacagggac aggaacctgc
     2821 ctcctctggc ccctctgggc ccccaccacc atcaccacca ttgagtctag acacgtgatt
     2881 aattaaggat cccccgacct cgacctctgg ctaataaagg aaatttattt tcattgcaat
     2941 agtgtgttgg aattttttgt gtctctcact cggaaggaca tatgggaggg caaatcattt
     3001 ggtcgagatc cctcggagat ctctagctag aggatcgatc cccgccccgg acgaactaaa
     3061 cctgactacg acatctctgc cccttcttcg cggggcagtg catgtaatcc cttcagttgg
     3121 ttggtacaac ttgccaactg ggccctgttc cacatgtgac acgggggggg accaaacaca
     3181 aaggggttct ctgactgtag ttgacatcct tataaatgga tgtgcacatt tgccaacact
     3241 gagtggcttt catcctggag cagactttgc agtctgtgga ctgcaacaca acattgcctt
     3301 tatgtgtaac tcttggctga agctcttaca ccaatgctgg gggacatgta cctcccaggg
     3361 gcccaggaag actacgggag gctacaccaa cgtcaatcag aggggcctgt gtagctaccg
     3421 ataagcggac cctcaagagg gcattagcaa tagtgtttat aaggccccct tgttaaccct
     3481 aaacgggtag catatgcttc ccgggtagta gtatatacta tccagactaa ccctaattca
     3541 atagcatatg ttacccaacg ggaagcatat gctatcgaat tagggttagt aaaagggtcc
     3601 taaggaacag cgatatctcc caccccatga gctgtcacgg ttttatttac atggggtcag
     3661 gattccacga gggtagtgaa ccattttagt cacaagggca gtggctgaag atcaaggagc
     3721 gggcagtgaa ctctcctgaa tcttcgcctg cttcttcatt ctccttcgtt tagctaatag
     3781 aataactgct gagttgtgaa cagtaaggtg tatgtgaggt gctcgaaaac aaggtttcag
     3841 gtgacgcccc cagaataaaa tttggacggg gggttcagtg gtggcattgt gctatgacac
     3901 caatataacc ctcacaaacc ccttgggcaa taaatactag tgtaggaatg aaacattctg
     3961 aatatcttta acaatagaaa tccatggggt ggggacaagc cgtaaagact ggatgtccat
     4021 ctcacacgaa tttatggcta tgggcaacac ataatcctag tgcaatatga tactggggtt
     4081 attaagatgt gtcccaggca gggaccaaga caggtgaacc atgttgttac actctatttg
     4141 taacaagggg aaagagagtg gacgccgaca gcagcggact ccactggttg tctctaacac
     4201 ccccgaaaat taaacggggc tccacgccaa tggggcccat aaacaaagac aagtggccac
     4261 tctttttttt gaaattgtgg agtgggggca cgcgtcagcc cccacacgcc gccctgcggt
     4321 tttggactgt aaaataaggg tgtaataact tggctgattg taaccccgct aaccactgcg
     4381 gtcaaaccac ttgcccacaa aaccactaat ggcaccccgg ggaatacctg cataagtagg
     4441 tgggcgggcc aagatagggg cgcgattgct gcgatctgga ggacaaatta cacacacttg
     4501 cgcctgagcg ccaagcacag ggttgttggt cctcatattc acgaggtcgc tgagagcacg
     4561 gtgggctaat gttgccatgg gtagcatata ctacccaaat atctggatag catatgctat
     4621 cctaatctat atctgggtag cataggctat cctaatctat atctgggtag catatgctat
     4681 cctaatctat atctgggtag tatatgctat cctaatttat atctgggtag cataggctat
     4741 cctaatctat atctgggtag catatgctat cctaatctat atctgggtag tatatgctat
     4801 cctaatctgt atccgggtag catatgctat cctaatagag attagggtag tatatgctat
     4861 cctaatttat atctgggtag catatactac ccaaatatct ggatagcata tgctatccta
     4921 atctatatct gggtagcata tgctatccta atctatatct gggtagcata ggctatccta
     4981 atctatatct gggtagcata tgctatccta atctatatct gggtagtata tgctatccta
     5041 atttatatct gggtagcata ggctatccta atctatatct gggtagcata tgctatccta
     5101 atctatatct gggtagtata tgctatccta atctgtatcc gggtagcata tgctatcctc
     5161 atgataagct gtcaaacatg agaattttct tgaagacgaa agggcctcgt gatacgccta
     5221 tttttatagg ttaatgtcat gataataatg gtttcttaga cgtcaggtgg cacttttcgg
     5281 ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg
     5341 ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtatgagt
     5401 attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt
     5461 gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg
     5521 ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa
     5581 cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtgtt
     5641 gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag
     5701 tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt
     5761 gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac gatcggagga
     5821 ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt
     5881 tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgca
     5941 gcaatggcaa caacgttgcg caaactatta actggcgaac tacttactct agcttcccgg
     6001 caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc
     6061 cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt
     6121 atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg
     6181 gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg
     6241 attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat tgatttaaaa
     6301 cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa
     6361 atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga
     6421 tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg
     6481 ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact
     6541 ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac
     6601 cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg
     6661 gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg
     6721 gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga
     6781 acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc
     6841 gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg
     6901 agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc
     6961 tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc
     7021 agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt
     7081 cctgcgttat cccctgattc tgtggataac cgtattaccg cctttgagtg agctgatacc
     7141 gctcgccgca gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc ggaagagcgc
     7201 ccaatacgca aaccgcctct ccccgcgcgt tggccgattc attaatgcag ctggcacgac
     7261 aggtttcccg actggaaagc gggcagtgag cgcaacgcaa ttaatgtgag ttagctcact
     7321 cattaggcac cccaggcttt acactttatg cttccggctc gtatgttgtg tggaattgtg
     7381 agcggataac aatttcacac aggaaacagc tatgaccatg attacgccaa gctctagcta
     7441 gaggtcgacc aattctcatg tttgacagct tatcatcgca gatccgggca acgttgttgc
     7501 cattgctgca ggcgcagaac tggtaggtat ggaagatcta tacattgaat caatattggc
     7561 aattagccat attagtcatt ggttatatag cataaatcaa tattggctat tggccattgc
     7621 atacgttgta tctatatcat aatatgtaca tttatattgg ctcatgtcca atatgaccgc
     7681 catgtt
//
