LOCUS       sequence-245744-        7803 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 7803)
  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 7803)
  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_P41-Cd4-bio-His
FEATURES             Location/Qualifiers
     source          1..7803
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..204
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     155..175
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     201..225
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     primer_bind     706..725
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     regulatory      748..757
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2581..2598
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    2650..2705
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(2704..2723)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(4941..4959)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5027..5131
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5132..5200,5201..5992)
                     /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(5350..5369)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6163..6751
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6652..6671
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6905..6922
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    7039..7060
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7075..7080,7081..7098,7099..7105)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7113..7129
                     /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     7118..7140
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7137..7153
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7137..7153
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     enhancer        7424..7803
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 gtgatgcggt tttggcagta caccaatggg cgtggatagc ggtttgactc acggggattt
       61 ccaagtctcc accccattga cgtcaatggg agtttgtttt ggcaccaaaa tcaacgggac
      121 tttccaaaat gtcgtaataa ccccgccccg ttgacgcaaa tgggcggtag gcgtgtacgg
      181 tgggaggtct atataagcag agctcgttta gtgaaccgtc agatcctcac tctcttccgc
      241 atcgctgtct gcgagggcca gctgttgggc tcgcggttga ggacaaactc ttcgcggtct
      301 ttccagtact cttggatcgg aaacccgtcg gcctccgaac ggtactccgc caccgaggga
      361 cctgagcgag tccgcatcga ccggatcgga aaacctctcg agaaaggcgt ctaaccagtc
      421 acagtcgcaa ggtaggctga gcaccgtggc gggcggcagc gggtggcggt cggggttgtt
      481 tctggcggag gtgctgctga tgatgtaatt aaagtaggcg gtcttgagac ggcggatggt
      541 cgaggtgagg tgtggcaggc ttgagatcca gctgttgggg tgagtactcc ctctcaaaag
      601 cgggcattac ttctgcgcta agattgtcag tttccaaaaa cgaggaggat ttgatattca
      661 cctggcccga tctggccata cacttgagtg acaatgacat ccactttgcc tttctctcca
      721 caggtgtcca ctcccaggtc caagtttgcc accatggagt ttcagaccca ggtactcatg
      781 tccctgctgc tctgcatgtc tggtgcggcc gccgaggaac acatctgcga cttcaccaaa
      841 gagaagtacc tgctgggcaa gaacgagaaa gaatactgcg tggtggaagc caagcccttc
      901 gacagcgtga ccttcatctg ccccaagaaa atcggcgccc agtgcttcca gaacgtgaac
      961 accgaggacg acatcagcaa agaaaagatg gaaagcagca agctggccat cgacgagctg
     1021 ctgtacggca gcaccctgta cggcgatacc ctgctgatct cccctaccgt gaagcagagc
     1081 aagaccttct actgcttctg caacctcgag atgaaggaca tgaagaagta tctgcagaag
     1141 aagcggctgt ctaaagagaa ggaaaaagag aaagagaacg ccaagaaaaa gtccaccgtg
     1201 aacgtggaca acctggaaaa cgccgacgag gacatggaag tgatcgtgcc ccagaagcag
     1261 gaagatcaga acctcgtccg ggccctgcac agagtgcgca agatcatgaa catcatcgag
     1321 cgcgagaaga acaaagagga agaaggcgac gactccgaga aaaagaaaga agaggacaaa
     1381 gaagaggaag aggaagaaga agaagaggaa gaactggtcg tcgaggaaaa agaggaagaa
     1441 aaggacaaga acaagatcga gaagatcatc accaaatacg gcgtgatgaa ggtggtggtg
     1501 tccaccaacc acgccatcac caagggctgc gatttcggca acaacgtggt caactacttc
     1561 agcaagccct accccatcga gagatacggc ggcagcaaag tgtgccggat cgaggccaag
     1621 cctggcgagt tcgtgggctt caagtgcatc tacgacaaca ccggcagcgt ggaaccccac
     1681 aactgcttcg acaaggtgtt ctacgagggc aaagaaaccg acctgcagac cctgatgccc
     1741 ggctacatca gctacggcaa caagaagaag ggcaaatacg ccttctacct gaagctgccc
     1801 cacttcgtgc agcacagcta caccatccgg tgcaagtgcc gcagcaccgt gtggcagtac
     1861 gacgactacg tgttcgagct ggccgtggaa ggcggcgagt ctgatgtggt ggccaagagc
     1921 ttccacgaag gcgcgccgtc gacctccatc acggcctata agagtgaggg ggagtcagcg
     1981 gagttctcct tcccactcaa ccttggagag gaaagcctgc agggagagtt gagatggaag
     2041 gcagagaagg ctccttcttc ccagtcctgg atcaccttct ccctaaagaa ccaaaaggtg
     2101 tctgtgcaga agtctactag caaccccaag ttccagctgt ccgaaacgct cccactcacc
     2161 cttcagatac cccaggtctc ccttcagttt gctggttctg gcaacctgac cctgactctg
     2221 gacagaggga tactgtatca ggaagtgaac ctggtggtga tgaaagtgac tcagcccgac
     2281 agcaacactt tgacctgtga ggtgatggga cccacctcac ccaagatgag actgatcttg
     2341 aagcaggaga atcaggaggc cagggtctcc aggcaggaga aagtgattca agtgcaggcc
     2401 cctgaagcag gggtgtggca atgtctactg agtgaaggtg aagaggtcaa gatggactcc
     2461 aagatccagg ttttatccaa agggttgaat tccggcagcc tgcaccacat cctggacgcc
     2521 cagaagatgc tgtggaacca cagggacagg aacctgcctc ctctggcccc tctgggcccc
     2581 caccaccatc accaccattg agtctagaca cgtgattaat taaggatccc ccgacctcga
     2641 cctctggcta ataaaggaaa tttattttca ttgcaatagt gtgttggaat tttttgtgtc
     2701 tctcactcgg aaggacatat gggagggcaa atcatttggt cgagatccct cggagatctc
     2761 tagctagagg atcgatcccc gccccggacg aactaaacct gactacgaca tctctgcccc
     2821 ttcttcgcgg ggcagtgcat gtaatccctt cagttggttg gtacaacttg ccaactgggc
     2881 cctgttccac atgtgacacg gggggggacc aaacacaaag gggttctctg actgtagttg
     2941 acatccttat aaatggatgt gcacatttgc caacactgag tggctttcat cctggagcag
     3001 actttgcagt ctgtggactg caacacaaca ttgcctttat gtgtaactct tggctgaagc
     3061 tcttacacca atgctggggg acatgtacct cccaggggcc caggaagact acgggaggct
     3121 acaccaacgt caatcagagg ggcctgtgta gctaccgata agcggaccct caagagggca
     3181 ttagcaatag tgtttataag gcccccttgt taaccctaaa cgggtagcat atgcttcccg
     3241 ggtagtagta tatactatcc agactaaccc taattcaata gcatatgtta cccaacggga
     3301 agcatatgct atcgaattag ggttagtaaa agggtcctaa ggaacagcga tatctcccac
     3361 cccatgagct gtcacggttt tatttacatg gggtcaggat tccacgaggg tagtgaacca
     3421 ttttagtcac aagggcagtg gctgaagatc aaggagcggg cagtgaactc tcctgaatct
     3481 tcgcctgctt cttcattctc cttcgtttag ctaatagaat aactgctgag ttgtgaacag
     3541 taaggtgtat gtgaggtgct cgaaaacaag gtttcaggtg acgcccccag aataaaattt
     3601 ggacgggggg ttcagtggtg gcattgtgct atgacaccaa tataaccctc acaaacccct
     3661 tgggcaataa atactagtgt aggaatgaaa cattctgaat atctttaaca atagaaatcc
     3721 atggggtggg gacaagccgt aaagactgga tgtccatctc acacgaattt atggctatgg
     3781 gcaacacata atcctagtgc aatatgatac tggggttatt aagatgtgtc ccaggcaggg
     3841 accaagacag gtgaaccatg ttgttacact ctatttgtaa caaggggaaa gagagtggac
     3901 gccgacagca gcggactcca ctggttgtct ctaacacccc cgaaaattaa acggggctcc
     3961 acgccaatgg ggcccataaa caaagacaag tggccactct tttttttgaa attgtggagt
     4021 gggggcacgc gtcagccccc acacgccgcc ctgcggtttt ggactgtaaa ataagggtgt
     4081 aataacttgg ctgattgtaa ccccgctaac cactgcggtc aaaccacttg cccacaaaac
     4141 cactaatggc accccgggga atacctgcat aagtaggtgg gcgggccaag ataggggcgc
     4201 gattgctgcg atctggagga caaattacac acacttgcgc ctgagcgcca agcacagggt
     4261 tgttggtcct catattcacg aggtcgctga gagcacggtg ggctaatgtt gccatgggta
     4321 gcatatacta cccaaatatc tggatagcat atgctatcct aatctatatc tgggtagcat
     4381 aggctatcct aatctatatc tgggtagcat atgctatcct aatctatatc tgggtagtat
     4441 atgctatcct aatttatatc tgggtagcat aggctatcct aatctatatc tgggtagcat
     4501 atgctatcct aatctatatc tgggtagtat atgctatcct aatctgtatc cgggtagcat
     4561 atgctatcct aatagagatt agggtagtat atgctatcct aatttatatc tgggtagcat
     4621 atactaccca aatatctgga tagcatatgc tatcctaatc tatatctggg tagcatatgc
     4681 tatcctaatc tatatctggg tagcataggc tatcctaatc tatatctggg tagcatatgc
     4741 tatcctaatc tatatctggg tagtatatgc tatcctaatt tatatctggg tagcataggc
     4801 tatcctaatc tatatctggg tagcatatgc tatcctaatc tatatctggg tagtatatgc
     4861 tatcctaatc tgtatccggg tagcatatgc tatcctcatg ataagctgtc aaacatgaga
     4921 attttcttga agacgaaagg gcctcgtgat acgcctattt ttataggtta atgtcatgat
     4981 aataatggtt tcttagacgt caggtggcac ttttcgggga aatgtgcgcg gaacccctat
     5041 ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata
     5101 aatgcttcaa taatattgaa aaaggaagag tatgagtatt caacatttcc gtgtcgccct
     5161 tattcccttt tttgcggcat tttgccttcc tgtttttgct cacccagaaa cgctggtgaa
     5221 agtaaaagat gctgaagatc agttgggtgc acgagtgggt tacatcgaac tggatctcaa
     5281 cagcggtaag atccttgaga gttttcgccc cgaagaacgt tttccaatga tgagcacttt
     5341 taaagttctg ctatgtggcg cggtattatc ccgtgttgac gccgggcaag agcaactcgg
     5401 tcgccgcata cactattctc agaatgactt ggttgagtac tcaccagtca cagaaaagca
     5461 tcttacggat ggcatgacag taagagaatt atgcagtgct gccataacca tgagtgataa
     5521 cactgcggcc aacttacttc tgacaacgat cggaggaccg aaggagctaa ccgctttttt
     5581 gcacaacatg ggggatcatg taactcgcct tgatcgttgg gaaccggagc tgaatgaagc
     5641 cataccaaac gacgagcgtg acaccacgat gcctgcagca atggcaacaa cgttgcgcaa
     5701 actattaact ggcgaactac ttactctagc ttcccggcaa caattaatag actggatgga
     5761 ggcggataaa gttgcaggac cacttctgcg ctcggccctt ccggctggct ggtttattgc
     5821 tgataaatct ggagccggtg agcgtgggtc tcgcggtatc attgcagcac tggggccaga
     5881 tggtaagccc tcccgtatcg tagttatcta cacgacgggg agtcaggcaa ctatggatga
     5941 acgaaataga cagatcgctg agataggtgc ctcactgatt aagcattggt aactgtcaga
     6001 ccaagtttac tcatatatac tttagattga tttaaaactt catttttaat ttaaaaggat
     6061 ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt
     6121 ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct
     6181 gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc
     6241 ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc
     6301 aaatactgtt cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc
     6361 gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc
     6421 gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg
     6481 aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata
     6541 cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta
     6601 tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc
     6661 ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg
     6721 atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt
     6781 cctggccttt tgctggcctt ttgctcacat gttctttcct gcgttatccc ctgattctgt
     6841 ggataaccgt attaccgcct ttgagtgagc tgataccgct cgccgcagcc gaacgaccga
     6901 gcgcagcgag tcagtgagcg aggaagcgga agagcgccca atacgcaaac cgcctctccc
     6961 cgcgcgttgg ccgattcatt aatgcagctg gcacgacagg tttcccgact ggaaagcggg
     7021 cagtgagcgc aacgcaatta atgtgagtta gctcactcat taggcacccc aggctttaca
     7081 ctttatgctt ccggctcgta tgttgtgtgg aattgtgagc ggataacaat ttcacacagg
     7141 aaacagctat gaccatgatt acgccaagct ctagctagag gtcgaccaat tctcatgttt
     7201 gacagcttat catcgcagat ccgggcaacg ttgttgccat tgctgcaggc gcagaactgg
     7261 taggtatgga agatctatac attgaatcaa tattggcaat tagccatatt agtcattggt
     7321 tatatagcat aaatcaatat tggctattgg ccattgcata cgttgtatct atatcataat
     7381 atgtacattt atattggctc atgtccaata tgaccgccat gttgacattg attattgact
     7441 agttattaat agtaatcaat tacggggtca ttagttcata gcccatatat ggagttccgc
     7501 gttacataac ttacggtaaa tggcccgcct ggctgaccgc ccaacgaccc ccgcccattg
     7561 acgtcaataa tgacgtatgt tcccatagta acgccaatag ggactttcca ttgacgtcaa
     7621 tgggtggagt atttacggta aactgcccac ttggcagtac atcaagtgta tcatatgcca
     7681 agtccgcccc ctattgacgt caatgacggt aaatggcccg cctggcatta tgcccagtac
     7741 atgaccttac gggactttcc tacttggcag tacatctacg tattagtcat cgctattacc
     7801 atg
//
