LOCUS       sequence-242835-        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        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             2751..2768
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    2820..2875
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(2874..2893)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(5111..5129)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5197..5301
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5302..5370,5371..6162)
                     /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(5520..5539)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6333..6921
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6822..6841
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7075..7092
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    7209..7230
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7245..7250,7251..7268,7269..7275)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7283..7299
                     /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     7288..7310
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7307..7323
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7307..7323
                     /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 gcggccgcct
     1201 ttacccacac ctgtgacttc aacgacgagc tggtgctgga cttcgacgac cagcagatga
     1261 ccaacagaga gcggatctgc agcctgaagc ccgacgtgtt cgacaaggtg gtcatcaagt
     1321 gcggcagcga gaagaacaaa tacgagctgc tgcccaacaa ctgcttcgaa caggtgtaca
     1381 caagcaagag catcaagaac acccagaacc tggtgcagta cctgcacggc gtggccgcca
     1441 tcgtgaagcg gaagcagaac aagtacaaca gccgggacaa gcggaactac gacgacgtgt
     1501 ccttccgggt ggcccctaac atggactccg agaagaaggc catctactgc atctgccaga
     1561 acaaggccaa gatccgcgtg aagaagaagt ccggcctgat ctacgacaag gacatcttca
     1621 acaccggcat cgtggaagtg gtcatcccca ccctgcccca gagaatcgac ggctgcgact
     1681 tcacccagaa taccgccacc ctgttcacca agggctacga cgtgaacttc tacaagaaca
     1741 tcgagaacga ggacgacgtg atctgcaaag tgcgggccac cgagggcaag ttcatcggct
     1801 tcaagtgccc cagcaactac gccatcaagc ccgaagagtg ctttctgcag ggcttcaacc
     1861 tggccggcaa gaaagagcag ctgcagagca aagtgaatct ggccgacctg gtcatggacc
     1921 actacaacaa ggtgttctac gcccgggtgc ccgagaggat ctaccagagc atgaactttt
     1981 tctgcacctg tgtgctggaa gagaagcggc tggtggccca cttcgagttt atcgccacca
     2041 gcagcgacgg catcagcatg agcgaggctg gcggcatgaa cgtgctgcag ggcagatctg
     2101 gcgcgccgtc gacctccatc acggcctata agagtgaggg ggagtcagcg gagttctcct
     2161 tcccactcaa ccttggagag gaaagcctgc agggagagtt gagatggaag gcagagaagg
     2221 ctccttcttc ccagtcctgg atcaccttct ccctaaagaa ccaaaaggtg tctgtgcaga
     2281 agtctactag caaccccaag ttccagctgt ccgaaacgct cccactcacc cttcagatac
     2341 cccaggtctc ccttcagttt gctggttctg gcaacctgac cctgactctg gacagaggga
     2401 tactgtatca ggaagtgaac ctggtggtga tgaaagtgac tcagcccgac agcaacactt
     2461 tgacctgtga ggtgatggga cccacctcac ccaagatgag actgatcttg aagcaggaga
     2521 atcaggaggc cagggtctcc aggcaggaga aagtgattca agtgcaggcc cctgaagcag
     2581 gggtgtggca atgtctactg agtgaaggtg aagaggtcaa gatggactcc aagatccagg
     2641 ttttatccaa agggttgaat tccggcagcc tgcaccacat cctggacgcc cagaagatgc
     2701 tgtggaacca cagggacagg aacctgcctc ctctggcccc tctgggcccc caccaccatc
     2761 accaccattg agtctagaca cgtgattaat taaggatccc ccgacctcga cctctggcta
     2821 ataaaggaaa tttattttca ttgcaatagt gtgttggaat tttttgtgtc tctcactcgg
     2881 aaggacatat gggagggcaa atcatttggt cgagatccct cggagatctc tagctagagg
     2941 atcgatcccc gccccggacg aactaaacct gactacgaca tctctgcccc ttcttcgcgg
     3001 ggcagtgcat gtaatccctt cagttggttg gtacaacttg ccaactgggc cctgttccac
     3061 atgtgacacg gggggggacc aaacacaaag gggttctctg actgtagttg acatccttat
     3121 aaatggatgt gcacatttgc caacactgag tggctttcat cctggagcag actttgcagt
     3181 ctgtggactg caacacaaca ttgcctttat gtgtaactct tggctgaagc tcttacacca
     3241 atgctggggg acatgtacct cccaggggcc caggaagact acgggaggct acaccaacgt
     3301 caatcagagg ggcctgtgta gctaccgata agcggaccct caagagggca ttagcaatag
     3361 tgtttataag gcccccttgt taaccctaaa cgggtagcat atgcttcccg ggtagtagta
     3421 tatactatcc agactaaccc taattcaata gcatatgtta cccaacggga agcatatgct
     3481 atcgaattag ggttagtaaa agggtcctaa ggaacagcga tatctcccac cccatgagct
     3541 gtcacggttt tatttacatg gggtcaggat tccacgaggg tagtgaacca ttttagtcac
     3601 aagggcagtg gctgaagatc aaggagcggg cagtgaactc tcctgaatct tcgcctgctt
     3661 cttcattctc cttcgtttag ctaatagaat aactgctgag ttgtgaacag taaggtgtat
     3721 gtgaggtgct cgaaaacaag gtttcaggtg acgcccccag aataaaattt ggacgggggg
     3781 ttcagtggtg gcattgtgct atgacaccaa tataaccctc acaaacccct tgggcaataa
     3841 atactagtgt aggaatgaaa cattctgaat atctttaaca atagaaatcc atggggtggg
     3901 gacaagccgt aaagactgga tgtccatctc acacgaattt atggctatgg gcaacacata
     3961 atcctagtgc aatatgatac tggggttatt aagatgtgtc ccaggcaggg accaagacag
     4021 gtgaaccatg ttgttacact ctatttgtaa caaggggaaa gagagtggac gccgacagca
     4081 gcggactcca ctggttgtct ctaacacccc cgaaaattaa acggggctcc acgccaatgg
     4141 ggcccataaa caaagacaag tggccactct tttttttgaa attgtggagt gggggcacgc
     4201 gtcagccccc acacgccgcc ctgcggtttt ggactgtaaa ataagggtgt aataacttgg
     4261 ctgattgtaa ccccgctaac cactgcggtc aaaccacttg cccacaaaac cactaatggc
     4321 accccgggga atacctgcat aagtaggtgg gcgggccaag ataggggcgc gattgctgcg
     4381 atctggagga caaattacac acacttgcgc ctgagcgcca agcacagggt tgttggtcct
     4441 catattcacg aggtcgctga gagcacggtg ggctaatgtt gccatgggta gcatatacta
     4501 cccaaatatc tggatagcat atgctatcct aatctatatc tgggtagcat aggctatcct
     4561 aatctatatc tgggtagcat atgctatcct aatctatatc tgggtagtat atgctatcct
     4621 aatttatatc tgggtagcat aggctatcct aatctatatc tgggtagcat atgctatcct
     4681 aatctatatc tgggtagtat atgctatcct aatctgtatc cgggtagcat atgctatcct
     4741 aatagagatt agggtagtat atgctatcct aatttatatc tgggtagcat atactaccca
     4801 aatatctgga tagcatatgc tatcctaatc tatatctggg tagcatatgc tatcctaatc
     4861 tatatctggg tagcataggc tatcctaatc tatatctggg tagcatatgc tatcctaatc
     4921 tatatctggg tagtatatgc tatcctaatt tatatctggg tagcataggc tatcctaatc
     4981 tatatctggg tagcatatgc tatcctaatc tatatctggg tagtatatgc tatcctaatc
     5041 tgtatccggg tagcatatgc tatcctcatg ataagctgtc aaacatgaga attttcttga
     5101 agacgaaagg gcctcgtgat acgcctattt ttataggtta atgtcatgat aataatggtt
     5161 tcttagacgt caggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt
     5221 ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa
     5281 taatattgaa aaaggaagag tatgagtatt caacatttcc gtgtcgccct tattcccttt
     5341 tttgcggcat tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat
     5401 gctgaagatc agttgggtgc acgagtgggt tacatcgaac tggatctcaa cagcggtaag
     5461 atccttgaga gttttcgccc cgaagaacgt tttccaatga tgagcacttt taaagttctg
     5521 ctatgtggcg cggtattatc ccgtgttgac gccgggcaag agcaactcgg tcgccgcata
     5581 cactattctc agaatgactt ggttgagtac tcaccagtca cagaaaagca tcttacggat
     5641 ggcatgacag taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc
     5701 aacttacttc tgacaacgat cggaggaccg aaggagctaa ccgctttttt gcacaacatg
     5761 ggggatcatg taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac
     5821 gacgagcgtg acaccacgat gcctgcagca atggcaacaa cgttgcgcaa actattaact
     5881 ggcgaactac ttactctagc ttcccggcaa caattaatag actggatgga ggcggataaa
     5941 gttgcaggac cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct
     6001 ggagccggtg agcgtgggtc tcgcggtatc attgcagcac tggggccaga tggtaagccc
     6061 tcccgtatcg tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga
     6121 cagatcgctg agataggtgc ctcactgatt aagcattggt aactgtcaga ccaagtttac
     6181 tcatatatac tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag
     6241 atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg
     6301 tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc
     6361 tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag
     6421 ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtt
     6481 cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac
     6541 ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc
     6601 gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt
     6661 tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt
     6721 gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc
     6781 ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt
     6841 tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca
     6901 ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt
     6961 tgctggcctt ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt
     7021 attaccgcct ttgagtgagc tgataccgct cgccgcagcc gaacgaccga gcgcagcgag
     7081 tcagtgagcg aggaagcgga agagcgccca atacgcaaac cgcctctccc cgcgcgttgg
     7141 ccgattcatt aatgcagctg gcacgacagg tttcccgact ggaaagcggg cagtgagcgc
     7201 aacgcaatta atgtgagtta gctcactcat taggcacccc aggctttaca ctttatgctt
     7261 ccggctcgta tgttgtgtgg aattgtgagc ggataacaat ttcacacagg aaacagctat
     7321 gaccatgatt acgccaagct ctagctagag gtcgaccaat tctcatgttt gacagcttat
     7381 catcgcagat ccgggcaacg ttgttgccat tgctgcaggc gcagaactgg taggtatgga
     7441 agatctatac attgaatcaa tattggcaat tagccatatt agtcattggt tatatagcat
     7501 aaatcaatat tggctattgg ccattgcata cgttgtatct atatcataat atgtacattt
     7561 atattggctc atgtccaata tgaccgc
//
