LOCUS       sequence-245652-        7587 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 7587)
  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 7587)
  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_P12-Cd4-bio-His
FEATURES             Location/Qualifiers
     source          1..7587
                     /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             2745..2762
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    2814..2869
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(2868..2887)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(5105..5123)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5191..5295
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5296..5364,5365..6156)
                     /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(5514..5533)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6327..6915
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6816..6835
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7069..7086
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    7203..7224
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7239..7244,7245..7262,7263..7269)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7277..7293
                     /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     7282..7304
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7301..7317
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7301..7317
                     /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 gcctttaccc
     1201 acacctgtga cttcaacgac gagctggtgc tggacttcga cgaccagcag atgaccaaca
     1261 gagagcggat ctgcagcctg aagcccgacg tgttcgacaa ggtggtcatc aagtgcggca
     1321 gcgagaagaa caaatacgag ctgctgccca acaactgctt cgaacaggtg tacacaagca
     1381 agagcatcaa gaacacccag aacctggtgc agtacctgca cggcgtggcc gccatcgtga
     1441 agcggaagca gaacaagtac aacagccggg acaagcggaa ctacgacgac gtgtccttcc
     1501 gggtggcccc taacatggac tccgagaaga aggccatcta ctgcatctgc cagaacaagg
     1561 ccaagatccg cgtgaagaag aagtccggcc tgatctacga caaggacatc ttcaacaccg
     1621 gcatcgtgga agtggtcatc cccaccctgc cccagagaat cgacggctgc gacttcaccc
     1681 agaataccgc caccctgttc accaagggct acgacgtgaa cttctacaag aacatcgaga
     1741 acgaggacga cgtgatctgc aaagtgcggg ccaccgaggg caagttcatc ggcttcaagt
     1801 gccccagcaa ctacgccatc aagcccgaag agtgctttct gcagggcttc aacctggccg
     1861 gcaagaaaga gcagctgcag agcaaagtga atctggccga cctggtcatg gaccactaca
     1921 acaaggtgtt ctacgcccgg gtgcccgaga ggatctacca gagcatgaac tttttctgca
     1981 cctgtgtgct ggaagagaag cggctggtgg cccacttcga gtttatcgcc accagcagcg
     2041 acggcatcag catgagcgag gctggcggca tgaacgtgct gcagggcaga tctggcgcgc
     2101 cgtcgacctc catcacggcc tataagagtg agggggagtc agcggagttc tccttcccac
     2161 tcaaccttgg agaggaaagc ctgcagggag agttgagatg gaaggcagag aaggctcctt
     2221 cttcccagtc ctggatcacc ttctccctaa agaaccaaaa ggtgtctgtg cagaagtcta
     2281 ctagcaaccc caagttccag ctgtccgaaa cgctcccact cacccttcag ataccccagg
     2341 tctcccttca gtttgctggt tctggcaacc tgaccctgac tctggacaga gggatactgt
     2401 atcaggaagt gaacctggtg gtgatgaaag tgactcagcc cgacagcaac actttgacct
     2461 gtgaggtgat gggacccacc tcacccaaga tgagactgat cttgaagcag gagaatcagg
     2521 aggccagggt ctccaggcag gagaaagtga ttcaagtgca ggcccctgaa gcaggggtgt
     2581 ggcaatgtct actgagtgaa ggtgaagagg tcaagatgga ctccaagatc caggttttat
     2641 ccaaagggtt gaattccggc agcctgcacc acatcctgga cgcccagaag atgctgtgga
     2701 accacaggga caggaacctg cctcctctgg cccctctggg cccccaccac catcaccacc
     2761 attgagtcta gacacgtgat taattaagga tcccccgacc tcgacctctg gctaataaag
     2821 gaaatttatt ttcattgcaa tagtgtgttg gaattttttg tgtctctcac tcggaaggac
     2881 atatgggagg gcaaatcatt tggtcgagat ccctcggaga tctctagcta gaggatcgat
     2941 ccccgccccg gacgaactaa acctgactac gacatctctg ccccttcttc gcggggcagt
     3001 gcatgtaatc ccttcagttg gttggtacaa cttgccaact gggccctgtt ccacatgtga
     3061 cacggggggg gaccaaacac aaaggggttc tctgactgta gttgacatcc ttataaatgg
     3121 atgtgcacat ttgccaacac tgagtggctt tcatcctgga gcagactttg cagtctgtgg
     3181 actgcaacac aacattgcct ttatgtgtaa ctcttggctg aagctcttac accaatgctg
     3241 ggggacatgt acctcccagg ggcccaggaa gactacggga ggctacacca acgtcaatca
     3301 gaggggcctg tgtagctacc gataagcgga ccctcaagag ggcattagca atagtgttta
     3361 taaggccccc ttgttaaccc taaacgggta gcatatgctt cccgggtagt agtatatact
     3421 atccagacta accctaattc aatagcatat gttacccaac gggaagcata tgctatcgaa
     3481 ttagggttag taaaagggtc ctaaggaaca gcgatatctc ccaccccatg agctgtcacg
     3541 gttttattta catggggtca ggattccacg agggtagtga accattttag tcacaagggc
     3601 agtggctgaa gatcaaggag cgggcagtga actctcctga atcttcgcct gcttcttcat
     3661 tctccttcgt ttagctaata gaataactgc tgagttgtga acagtaaggt gtatgtgagg
     3721 tgctcgaaaa caaggtttca ggtgacgccc ccagaataaa atttggacgg ggggttcagt
     3781 ggtggcattg tgctatgaca ccaatataac cctcacaaac cccttgggca ataaatacta
     3841 gtgtaggaat gaaacattct gaatatcttt aacaatagaa atccatgggg tggggacaag
     3901 ccgtaaagac tggatgtcca tctcacacga atttatggct atgggcaaca cataatccta
     3961 gtgcaatatg atactggggt tattaagatg tgtcccaggc agggaccaag acaggtgaac
     4021 catgttgtta cactctattt gtaacaaggg gaaagagagt ggacgccgac agcagcggac
     4081 tccactggtt gtctctaaca cccccgaaaa ttaaacgggg ctccacgcca atggggccca
     4141 taaacaaaga caagtggcca ctcttttttt tgaaattgtg gagtgggggc acgcgtcagc
     4201 ccccacacgc cgccctgcgg ttttggactg taaaataagg gtgtaataac ttggctgatt
     4261 gtaaccccgc taaccactgc ggtcaaacca cttgcccaca aaaccactaa tggcaccccg
     4321 gggaatacct gcataagtag gtgggcgggc caagataggg gcgcgattgc tgcgatctgg
     4381 aggacaaatt acacacactt gcgcctgagc gccaagcaca gggttgttgg tcctcatatt
     4441 cacgaggtcg ctgagagcac ggtgggctaa tgttgccatg ggtagcatat actacccaaa
     4501 tatctggata gcatatgcta tcctaatcta tatctgggta gcataggcta tcctaatcta
     4561 tatctgggta gcatatgcta tcctaatcta tatctgggta gtatatgcta tcctaattta
     4621 tatctgggta gcataggcta tcctaatcta tatctgggta gcatatgcta tcctaatcta
     4681 tatctgggta gtatatgcta tcctaatctg tatccgggta gcatatgcta tcctaataga
     4741 gattagggta gtatatgcta tcctaattta tatctgggta gcatatacta cccaaatatc
     4801 tggatagcat atgctatcct aatctatatc tgggtagcat atgctatcct aatctatatc
     4861 tgggtagcat aggctatcct aatctatatc tgggtagcat atgctatcct aatctatatc
     4921 tgggtagtat atgctatcct aatttatatc tgggtagcat aggctatcct aatctatatc
     4981 tgggtagcat atgctatcct aatctatatc tgggtagtat atgctatcct aatctgtatc
     5041 cgggtagcat atgctatcct catgataagc tgtcaaacat gagaattttc ttgaagacga
     5101 aagggcctcg tgatacgcct atttttatag gttaatgtca tgataataat ggtttcttag
     5161 acgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa
     5221 atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat
     5281 tgaaaaagga agagtatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg
     5341 gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa
     5401 gatcagttgg gtgcacgagt gggttacatc gaactggatc tcaacagcgg taagatcctt
     5461 gagagttttc gccccgaaga acgttttcca atgatgagca cttttaaagt tctgctatgt
     5521 ggcgcggtat tatcccgtgt tgacgccggg caagagcaac tcggtcgccg catacactat
     5581 tctcagaatg acttggttga gtactcacca gtcacagaaa agcatcttac ggatggcatg
     5641 acagtaagag aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta
     5701 cttctgacaa cgatcggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat
     5761 catgtaactc gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag
     5821 cgtgacacca cgatgcctgc agcaatggca acaacgttgc gcaaactatt aactggcgaa
     5881 ctacttactc tagcttcccg gcaacaatta atagactgga tggaggcgga taaagttgca
     5941 ggaccacttc tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc
     6001 ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt
     6061 atcgtagtta tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc
     6121 gctgagatag gtgcctcact gattaagcat tggtaactgt cagaccaagt ttactcatat
     6181 atactttaga ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt
     6241 tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac
     6301 cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc
     6361 ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca
     6421 actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta
     6481 gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct
     6541 ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg
     6601 gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc
     6661 acacagccca gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta
     6721 tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg
     6781 gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt
     6841 cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg
     6901 cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg
     6961 ccttttgctc acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc
     7021 gcctttgagt gagctgatac cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg
     7081 agcgaggaag cggaagagcg cccaatacgc aaaccgcctc tccccgcgcg ttggccgatt
     7141 cattaatgca gctggcacga caggtttccc gactggaaag cgggcagtga gcgcaacgca
     7201 attaatgtga gttagctcac tcattaggca ccccaggctt tacactttat gcttccggct
     7261 cgtatgttgt gtggaattgt gagcggataa caatttcaca caggaaacag ctatgaccat
     7321 gattacgcca agctctagct agaggtcgac caattctcat gtttgacagc ttatcatcgc
     7381 agatccgggc aacgttgttg ccattgctgc aggcgcagaa ctggtaggta tggaagatct
     7441 atacattgaa tcaatattgg caattagcca tattagtcat tggttatata gcataaatca
     7501 atattggcta ttggccattg catacgttgt atctatatca taatatgtac atttatattg
     7561 gctcatgtcc aatatgaccg ccatgtt
//
