LOCUS       sequence-245756-        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"
     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             2638..2655
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    2707..2762
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(2761..2780)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(4998..5016)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5084..5188
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5189..5257,5258..6049)
                     /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(5407..5426)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6220..6808
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6709..6728
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6962..6979
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    7096..7117
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7132..7137,7138..7155,7156..7162)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7170..7186
                     /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     7175..7197
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7194..7210
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7194..7210
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     enhancer        7481..7860
                     /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 gccaatggca atgagctgaa gggcacagat
      841 ggcgtggacc ctcccaagca ccaggacatc accaacgact acaacaccct ggaacacatc
      901 agcagcgagg gccagatcag ccggcggaac gacgatagca ccatccctca gagcaacacc
      961 cagcaggaag aggaaggcag cgacgtcaaa gagggcaaca ccttcctgca cggcaagaac
     1021 gtggaaaaag aggtggaaac ctacctgatc caggaagaac agaacaaagt caaccacgcc
     1081 gacctgggca ccggcaagca ggaaaaagcc agccaccacg tgcagaagaa cgccaacctg
     1141 cggagcacca gcaagagcag ctctgaggcc gccgtgaacc aggcctacaa cttcgtgcag
     1201 cggaacagcg cccctctggg caagaacaag gccaacatcg aacaggtggc cagagcccag
     1261 gacgacatcg acaccaccga gaacgacaac acccacgaca ccaaagagca caacgccgac
     1321 gccgacaacc tgaacggcga gggcgataac gccgagaagg gcaataccga gaatgccgac
     1381 gccaagaagg acaacgctga ggacgccaac gccgaaaagg atatcgccaa gaatgtgaac
     1441 gcagagaagg acattgccaa aaacgccaat gccgataccg gcaagggcgg cgaggaacac
     1501 gaggaaccca gaatcagccc tcccgccgag cagcctacca gaaacgcccc tcctctggac
     1561 catgccgaga agatcaagaa ccggctgctg aaagagggcc gcgagctgaa agaaaccacc
     1621 agcatgatcg acaataccgt gtacaacgtc gagcagatca tcctcaagac caagttctac
     1681 accaccgcca tccggaactt cgtcctgttc aaagtgaacc acatctgcga gtacagcaag
     1741 tgcggcccca acgcccggtg ctacatcgtg gaaaaggaca aagaggaatg ccgctgtatc
     1801 gccaactaca tgcccgacaa cagcgtggac tacttcaagt gcatccccaa gaccgtgaag
     1861 gactgcagca aagagaacgg caactgcgac gtgaacgccg agtgcagcat cgacaagaaa
     1921 gaaaacatca agtgccagtg caaccacggc tatatcggcg acggcatctt ctgcgtgctg
     1981 ggctctggcg cgccgtcgac ctccatcacg gcctataaga gtgaggggga gtcagcggag
     2041 ttctccttcc cactcaacct tggagaggaa agcctgcagg gagagttgag atggaaggca
     2101 gagaaggctc cttcttccca gtcctggatc accttctccc taaagaacca aaaggtgtct
     2161 gtgcagaagt ctactagcaa ccccaagttc cagctgtccg aaacgctccc actcaccctt
     2221 cagatacccc aggtctccct tcagtttgct ggttctggca acctgaccct gactctggac
     2281 agagggatac tgtatcagga agtgaacctg gtggtgatga aagtgactca gcccgacagc
     2341 aacactttga cctgtgaggt gatgggaccc acctcaccca agatgagact gatcttgaag
     2401 caggagaatc aggaggccag ggtctccagg caggagaaag tgattcaagt gcaggcccct
     2461 gaagcagggg tgtggcaatg tctactgagt gaaggtgaag aggtcaagat ggactccaag
     2521 atccaggttt tatccaaagg gttgaattcc ggcagcctgc accacatcct ggacgcccag
     2581 aagatgctgt ggaaccacag ggacaggaac ctgcctcctc tggcccctct gggcccccac
     2641 caccatcacc accattgagt ctagacacgt gattaattaa ggatcccccg acctcgacct
     2701 ctggctaata aaggaaattt attttcattg caatagtgtg ttggaatttt ttgtgtctct
     2761 cactcggaag gacatatggg agggcaaatc atttggtcga gatccctcgg agatctctag
     2821 ctagaggatc gatccccgcc ccggacgaac taaacctgac tacgacatct ctgccccttc
     2881 ttcgcggggc agtgcatgta atcccttcag ttggttggta caacttgcca actgggccct
     2941 gttccacatg tgacacgggg ggggaccaaa cacaaagggg ttctctgact gtagttgaca
     3001 tccttataaa tggatgtgca catttgccaa cactgagtgg ctttcatcct ggagcagact
     3061 ttgcagtctg tggactgcaa cacaacattg cctttatgtg taactcttgg ctgaagctct
     3121 tacaccaatg ctgggggaca tgtacctccc aggggcccag gaagactacg ggaggctaca
     3181 ccaacgtcaa tcagaggggc ctgtgtagct accgataagc ggaccctcaa gagggcatta
     3241 gcaatagtgt ttataaggcc cccttgttaa ccctaaacgg gtagcatatg cttcccgggt
     3301 agtagtatat actatccaga ctaaccctaa ttcaatagca tatgttaccc aacgggaagc
     3361 atatgctatc gaattagggt tagtaaaagg gtcctaagga acagcgatat ctcccacccc
     3421 atgagctgtc acggttttat ttacatgggg tcaggattcc acgagggtag tgaaccattt
     3481 tagtcacaag ggcagtggct gaagatcaag gagcgggcag tgaactctcc tgaatcttcg
     3541 cctgcttctt cattctcctt cgtttagcta atagaataac tgctgagttg tgaacagtaa
     3601 ggtgtatgtg aggtgctcga aaacaaggtt tcaggtgacg cccccagaat aaaatttgga
     3661 cggggggttc agtggtggca ttgtgctatg acaccaatat aaccctcaca aaccccttgg
     3721 gcaataaata ctagtgtagg aatgaaacat tctgaatatc tttaacaata gaaatccatg
     3781 gggtggggac aagccgtaaa gactggatgt ccatctcaca cgaatttatg gctatgggca
     3841 acacataatc ctagtgcaat atgatactgg ggttattaag atgtgtccca ggcagggacc
     3901 aagacaggtg aaccatgttg ttacactcta tttgtaacaa ggggaaagag agtggacgcc
     3961 gacagcagcg gactccactg gttgtctcta acacccccga aaattaaacg gggctccacg
     4021 ccaatggggc ccataaacaa agacaagtgg ccactctttt ttttgaaatt gtggagtggg
     4081 ggcacgcgtc agcccccaca cgccgccctg cggttttgga ctgtaaaata agggtgtaat
     4141 aacttggctg attgtaaccc cgctaaccac tgcggtcaaa ccacttgccc acaaaaccac
     4201 taatggcacc ccggggaata cctgcataag taggtgggcg ggccaagata ggggcgcgat
     4261 tgctgcgatc tggaggacaa attacacaca cttgcgcctg agcgccaagc acagggttgt
     4321 tggtcctcat attcacgagg tcgctgagag cacggtgggc taatgttgcc atgggtagca
     4381 tatactaccc aaatatctgg atagcatatg ctatcctaat ctatatctgg gtagcatagg
     4441 ctatcctaat ctatatctgg gtagcatatg ctatcctaat ctatatctgg gtagtatatg
     4501 ctatcctaat ttatatctgg gtagcatagg ctatcctaat ctatatctgg gtagcatatg
     4561 ctatcctaat ctatatctgg gtagtatatg ctatcctaat ctgtatccgg gtagcatatg
     4621 ctatcctaat agagattagg gtagtatatg ctatcctaat ttatatctgg gtagcatata
     4681 ctacccaaat atctggatag catatgctat cctaatctat atctgggtag catatgctat
     4741 cctaatctat atctgggtag cataggctat cctaatctat atctgggtag catatgctat
     4801 cctaatctat atctgggtag tatatgctat cctaatttat atctgggtag cataggctat
     4861 cctaatctat atctgggtag catatgctat cctaatctat atctgggtag tatatgctat
     4921 cctaatctgt atccgggtag catatgctat cctcatgata agctgtcaaa catgagaatt
     4981 ttcttgaaga cgaaagggcc tcgtgatacg cctattttta taggttaatg tcatgataat
     5041 aatggtttct tagacgtcag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg
     5101 tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat
     5161 gcttcaataa tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat
     5221 tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt
     5281 aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag
     5341 cggtaagatc cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa
     5401 agttctgcta tgtggcgcgg tattatcccg tgttgacgcc gggcaagagc aactcggtcg
     5461 ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct
     5521 tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac
     5581 tgcggccaac ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca
     5641 caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat
     5701 accaaacgac gagcgtgaca ccacgatgcc tgcagcaatg gcaacaacgt tgcgcaaact
     5761 attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc
     5821 ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga
     5881 taaatctgga gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg
     5941 taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg
     6001 aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca
     6061 agtttactca tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta
     6121 ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca
     6181 ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg
     6241 cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga
     6301 tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa
     6361 tactgttctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc
     6421 tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg
     6481 tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac
     6541 ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct
     6601 acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc
     6661 ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg
     6721 gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg
     6781 ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct
     6841 ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga
     6901 taaccgtatt accgcctttg agtgagctga taccgctcgc cgcagccgaa cgaccgagcg
     6961 cagcgagtca gtgagcgagg aagcggaaga gcgcccaata cgcaaaccgc ctctccccgc
     7021 gcgttggccg attcattaat gcagctggca cgacaggttt cccgactgga aagcgggcag
     7081 tgagcgcaac gcaattaatg tgagttagct cactcattag gcaccccagg ctttacactt
     7141 tatgcttccg gctcgtatgt tgtgtggaat tgtgagcgga taacaatttc acacaggaaa
     7201 cagctatgac catgattacg ccaagctcta gctagaggtc gaccaattct catgtttgac
     7261 agcttatcat cgcagatccg ggcaacgttg ttgccattgc tgcaggcgca gaactggtag
     7321 gtatggaaga tctatacatt gaatcaatat tggcaattag ccatattagt cattggttat
     7381 atagcataaa tcaatattgg ctattggcca ttgcatacgt tgtatctata tcataatatg
     7441 tacatttata ttggctcatg tccaatatga ccgccatgtt gacattgatt attgactagt
     7501 tattaatagt aatcaattac ggggtcatta gttcatagcc catatatgga gttccgcgtt
     7561 acataactta cggtaaatgg cccgcctggc tgaccgccca acgacccccg cccattgacg
     7621 tcaataatga cgtatgttcc catagtaacg ccaataggga ctttccattg acgtcaatgg
     7681 gtggagtatt tacggtaaac tgcccacttg gcagtacatc aagtgtatca tatgccaagt
     7741 ccgcccccta ttgacgtcaa tgacggtaaa tggcccgcct ggcattatgc ccagtacatg
     7801 accttacggg actttcctac ttggcagtac atctacgtat tagtcatcgc tattaccatg
//
