LOCUS       sequence-245758-        9189 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 9189)
  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 9189)
  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_P92-Cd4-bio-His
FEATURES             Location/Qualifiers
     source          1..9189
                     /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             3967..3984
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    4036..4091
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(4090..4109)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(6327..6345)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6413..6517
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6518..6586,6587..7378)
                     /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(6736..6755)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7549..8137
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8038..8057
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     8291..8308
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    8425..8446
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(8461..8466,8467..8484,8485..8491)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8499..8515
                     /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     8504..8526
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     8523..8539
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     8523..8539
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     enhancer        8810..9189
                     /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 gccgatgtga tcctggaagg cgagtttgag
      841 agcaagctgg gctacgacgg cgagcagatc gtgtacagca agaagaactt caagggcagc
      901 atctacgcct tcgtgcccgt gaagggcaag tgcggcatca gcgtgcagga aagcacagac
      961 ggcgtcgagt gggagaacaa cgccgccatc gaggtggaag atggcgtgaa cgaggccacc
     1021 ctgttcagcg tgttcaccac cgacgacgcc gtgatcgtga tcttccggtg cagcaacatc
     1081 tactatatcg ccaagcgggg cgagggcaac gcctggaaga acatcaaaca ggtgtccttc
     1141 agcgacttcc ccgccgacgc catccctgcc gtgtactctg gcagctacct gctgatgaac
     1201 aacgagtacg agaagttcat cctcgtgtgc gccagagagg accccaacgg cgccgaacac
     1261 ggcaacgagg ccgatagcga agtgaaggcc gtgcctttca gcgaactgca gctgctgggc
     1321 cggtgcaagt tcagcttcga cgagggcgag acatggcgcg agggcgtgac cagaatccac
     1381 agcgacgacc tgagcagcta tgacggcggc gaggatagca ccgagctgag ccacgatgac
     1441 gccctgaagt ccgtgaaaga agtgaacatg gccgaggaag aggaagtcaa gctggaaggc
     1501 cagggcaccg aggaatccaa agagggcgct cccctgcgga acggcatgca gatcagactg
     1561 atccagatgg gcggcaacct ggccatccgg gccgtgaaca ttgccaagga cagcaacagc
     1621 gccaagaccg tgatccaggt ctgcaacagc ctgtacaaga acgacctgta ctgcgacaaa
     1681 gtgaagctgc agatcaagga cgagtacgtg ctgcacacct acgagatggt caacggctac
     1741 cacatggccg tgctgagaga gagaaacggc gcccacctgg tgcccagctt cgtgtacaat
     1801 ctggccgaga cattcgagcc catcctgcac tacaaatacg agaacaatgg ggagggcaag
     1861 agcgacgtcg tgggcggcat ccctctggac aagaccacct atgagttcgt gctggccaag
     1921 ggcgatatgg tgttcctgtt ctccagcacc gtggggaaga agaacaaagt catcaagatc
     1981 aacgcccctc ggcggaaggc cggatgcgaa gtcctgcctg atggcgagag caacaagggc
     2041 tacagccaca ccttcagcat ccggaacatg cagaaccacc ggatctgcaa gatggccacc
     2101 agcgagctgg gaaccaccga ggacggcctg ttcaaggtgt tctacgtgac cctgcccaag
     2161 ggcgtgtccc tgaacgacga ctgcttcaga ttcagcttca gcaaagagct gaagtctgag
     2221 taccacacca ccgtggtcac caagagcacc accggcaatg ccgctcacga aggggagatc
     2281 cagtttagct tcccactgta ctacagcaag tacctgtaca actacaagaa ggcctactgc
     2341 accctgagca acggcctgat ggtggccgtg gaattcgact acctgagaaa catcctggac
     2401 ctgaactacg ccgacgagaa cgacgtgatc aagattacca gcaacgacat cgtggtgcat
     2461 gccaagcgga gatacgtcca ggaaaagggc ggcgacatct tccccagcgg cacctacatc
     2521 aacacctacc tgcccttcga gaaagagtac gtcatcagca acttcgtccc caagtctgtg
     2581 aatgaggtgt ccaccatcac cctgccccac ggctctgtgc ggaagctgca ctttctgcag
     2641 agcggcgagg tgctgcccta cgatggcatc gacctgtcca acgtggcccc taattacaag
     2701 aagctgacta agtacgagct gaaccgcaag accctggatg tgatcatctc caacttcaag
     2761 tccgatgaga agattatcgg actcgtgtgc cctctgagca gcgaggatga gggcctgacc
     2821 tgcttcgaca aggtgtacct gcagaagggc ggcctgcaca acatcgagtt cgtgtttggc
     2881 agcaacgata tcttcgtgat ccctcagcgg cggctgctga ggcctggcgg caaaatggcc
     2941 atggaaagcc tgctgtacct gaacaagcgg aacgtggaca agctgaagaa agagaagaat
     3001 atcatccact ttttctgcga gtgcaagggc gacgaggaca ccgagagcgt gcgcgtgaac
     3061 tactacatca gcgcctacta cgacagcgag tttgtgaagg cccaggtcgg aaagtccggc
     3121 gagtccagct tcatcaacgg ccacatcatg gtgcagcggg aagaggccgg caaagaatcc
     3181 accgtggata aggtggacga caaggtcgac tggatcagca gcgacaccaa cctgaacatg
     3241 gaagagggcg tccggatgat cggccagaac atgctgttta gcatgctgag cggcggcaac
     3301 agcaagggca accctggcgc gccgtcgacc tccatcacgg cctataagag tgagggggag
     3361 tcagcggagt tctccttccc actcaacctt ggagaggaaa gcctgcaggg agagttgaga
     3421 tggaaggcag agaaggctcc ttcttcccag tcctggatca ccttctccct aaagaaccaa
     3481 aaggtgtctg tgcagaagtc tactagcaac cccaagttcc agctgtccga aacgctccca
     3541 ctcacccttc agatacccca ggtctccctt cagtttgctg gttctggcaa cctgaccctg
     3601 actctggaca gagggatact gtatcaggaa gtgaacctgg tggtgatgaa agtgactcag
     3661 cccgacagca acactttgac ctgtgaggtg atgggaccca cctcacccaa gatgagactg
     3721 atcttgaagc aggagaatca ggaggccagg gtctccaggc aggagaaagt gattcaagtg
     3781 caggcccctg aagcaggggt gtggcaatgt ctactgagtg aaggtgaaga ggtcaagatg
     3841 gactccaaga tccaggtttt atccaaaggg ttgaattccg gcagcctgca ccacatcctg
     3901 gacgcccaga agatgctgtg gaaccacagg gacaggaacc tgcctcctct ggcccctctg
     3961 ggcccccacc accatcacca ccattgagtc tagacacgtg attaattaag gatcccccga
     4021 cctcgacctc tggctaataa aggaaattta ttttcattgc aatagtgtgt tggaattttt
     4081 tgtgtctctc actcggaagg acatatggga gggcaaatca tttggtcgag atccctcgga
     4141 gatctctagc tagaggatcg atccccgccc cggacgaact aaacctgact acgacatctc
     4201 tgccccttct tcgcggggca gtgcatgtaa tcccttcagt tggttggtac aacttgccaa
     4261 ctgggccctg ttccacatgt gacacggggg gggaccaaac acaaaggggt tctctgactg
     4321 tagttgacat ccttataaat ggatgtgcac atttgccaac actgagtggc tttcatcctg
     4381 gagcagactt tgcagtctgt ggactgcaac acaacattgc ctttatgtgt aactcttggc
     4441 tgaagctctt acaccaatgc tgggggacat gtacctccca ggggcccagg aagactacgg
     4501 gaggctacac caacgtcaat cagaggggcc tgtgtagcta ccgataagcg gaccctcaag
     4561 agggcattag caatagtgtt tataaggccc ccttgttaac cctaaacggg tagcatatgc
     4621 ttcccgggta gtagtatata ctatccagac taaccctaat tcaatagcat atgttaccca
     4681 acgggaagca tatgctatcg aattagggtt agtaaaaggg tcctaaggaa cagcgatatc
     4741 tcccacccca tgagctgtca cggttttatt tacatggggt caggattcca cgagggtagt
     4801 gaaccatttt agtcacaagg gcagtggctg aagatcaagg agcgggcagt gaactctcct
     4861 gaatcttcgc ctgcttcttc attctccttc gtttagctaa tagaataact gctgagttgt
     4921 gaacagtaag gtgtatgtga ggtgctcgaa aacaaggttt caggtgacgc ccccagaata
     4981 aaatttggac ggggggttca gtggtggcat tgtgctatga caccaatata accctcacaa
     5041 accccttggg caataaatac tagtgtagga atgaaacatt ctgaatatct ttaacaatag
     5101 aaatccatgg ggtggggaca agccgtaaag actggatgtc catctcacac gaatttatgg
     5161 ctatgggcaa cacataatcc tagtgcaata tgatactggg gttattaaga tgtgtcccag
     5221 gcagggacca agacaggtga accatgttgt tacactctat ttgtaacaag gggaaagaga
     5281 gtggacgccg acagcagcgg actccactgg ttgtctctaa cacccccgaa aattaaacgg
     5341 ggctccacgc caatggggcc cataaacaaa gacaagtggc cactcttttt tttgaaattg
     5401 tggagtgggg gcacgcgtca gcccccacac gccgccctgc ggttttggac tgtaaaataa
     5461 gggtgtaata acttggctga ttgtaacccc gctaaccact gcggtcaaac cacttgccca
     5521 caaaaccact aatggcaccc cggggaatac ctgcataagt aggtgggcgg gccaagatag
     5581 gggcgcgatt gctgcgatct ggaggacaaa ttacacacac ttgcgcctga gcgccaagca
     5641 cagggttgtt ggtcctcata ttcacgaggt cgctgagagc acggtgggct aatgttgcca
     5701 tgggtagcat atactaccca aatatctgga tagcatatgc tatcctaatc tatatctggg
     5761 tagcataggc tatcctaatc tatatctggg tagcatatgc tatcctaatc tatatctggg
     5821 tagtatatgc tatcctaatt tatatctggg tagcataggc tatcctaatc tatatctggg
     5881 tagcatatgc tatcctaatc tatatctggg tagtatatgc tatcctaatc tgtatccggg
     5941 tagcatatgc tatcctaata gagattaggg tagtatatgc tatcctaatt tatatctggg
     6001 tagcatatac tacccaaata tctggatagc atatgctatc ctaatctata tctgggtagc
     6061 atatgctatc ctaatctata tctgggtagc ataggctatc ctaatctata tctgggtagc
     6121 atatgctatc ctaatctata tctgggtagt atatgctatc ctaatttata tctgggtagc
     6181 ataggctatc ctaatctata tctgggtagc atatgctatc ctaatctata tctgggtagt
     6241 atatgctatc ctaatctgta tccgggtagc atatgctatc ctcatgataa gctgtcaaac
     6301 atgagaattt tcttgaagac gaaagggcct cgtgatacgc ctatttttat aggttaatgt
     6361 catgataata atggtttctt agacgtcagg tggcactttt cggggaaatg tgcgcggaac
     6421 ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc
     6481 ctgataaatg cttcaataat attgaaaaag gaagagtatg agtattcaac atttccgtgt
     6541 cgcccttatt cccttttttg cggcattttg ccttcctgtt tttgctcacc cagaaacgct
     6601 ggtgaaagta aaagatgctg aagatcagtt gggtgcacga gtgggttaca tcgaactgga
     6661 tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc caatgatgag
     6721 cacttttaaa gttctgctat gtggcgcggt attatcccgt gttgacgccg ggcaagagca
     6781 actcggtcgc cgcatacact attctcagaa tgacttggtt gagtactcac cagtcacaga
     6841 aaagcatctt acggatggca tgacagtaag agaattatgc agtgctgcca taaccatgag
     6901 tgataacact gcggccaact tacttctgac aacgatcgga ggaccgaagg agctaaccgc
     6961 ttttttgcac aacatggggg atcatgtaac tcgccttgat cgttgggaac cggagctgaa
     7021 tgaagccata ccaaacgacg agcgtgacac cacgatgcct gcagcaatgg caacaacgtt
     7081 gcgcaaacta ttaactggcg aactacttac tctagcttcc cggcaacaat taatagactg
     7141 gatggaggcg gataaagttg caggaccact tctgcgctcg gcccttccgg ctggctggtt
     7201 tattgctgat aaatctggag ccggtgagcg tgggtctcgc ggtatcattg cagcactggg
     7261 gccagatggt aagccctccc gtatcgtagt tatctacacg acggggagtc aggcaactat
     7321 ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc attggtaact
     7381 gtcagaccaa gtttactcat atatacttta gattgattta aaacttcatt tttaatttaa
     7441 aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt
     7501 ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt
     7561 ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg
     7621 tttgccggat caagagctac caactctttt tccgaaggta actggcttca gcagagcgca
     7681 gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt
     7741 agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga
     7801 taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc
     7861 gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact
     7921 gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga
     7981 caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg
     8041 aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt
     8101 tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt
     8161 acggttcctg gccttttgct ggccttttgc tcacatgttc tttcctgcgt tatcccctga
     8221 ttctgtggat aaccgtatta ccgcctttga gtgagctgat accgctcgcc gcagccgaac
     8281 gaccgagcgc agcgagtcag tgagcgagga agcggaagag cgcccaatac gcaaaccgcc
     8341 tctccccgcg cgttggccga ttcattaatg cagctggcac gacaggtttc ccgactggaa
     8401 agcgggcagt gagcgcaacg caattaatgt gagttagctc actcattagg caccccaggc
     8461 tttacacttt atgcttccgg ctcgtatgtt gtgtggaatt gtgagcggat aacaatttca
     8521 cacaggaaac agctatgacc atgattacgc caagctctag ctagaggtcg accaattctc
     8581 atgtttgaca gcttatcatc gcagatccgg gcaacgttgt tgccattgct gcaggcgcag
     8641 aactggtagg tatggaagat ctatacattg aatcaatatt ggcaattagc catattagtc
     8701 attggttata tagcataaat caatattggc tattggccat tgcatacgtt gtatctatat
     8761 cataatatgt acatttatat tggctcatgt ccaatatgac cgccatgttg acattgatta
     8821 ttgactagtt attaatagta atcaattacg gggtcattag ttcatagccc atatatggag
     8881 ttccgcgtta cataacttac ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc
     8941 ccattgacgt caataatgac gtatgttccc atagtaacgc caatagggac tttccattga
     9001 cgtcaatggg tggagtattt acggtaaact gcccacttgg cagtacatca agtgtatcat
     9061 atgccaagtc cgccccctat tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc
     9121 cagtacatga ccttacggga ctttcctact tggcagtaca tctacgtatt agtcatcgct
     9181 attaccatg
//
