LOCUS       sequence-242837-        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"
     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             2967..2984
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    3036..3091
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(3090..3109)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(5327..5345)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5413..5517
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5518..5586,5587..6378)
                     /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(5736..5755)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6549..7137
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7038..7057
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7291..7308
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    7425..7446
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7461..7466,7467..7484,7485..7491)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7499..7515
                     /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     7504..7526
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7523..7539
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7523..7539
                     /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 gcggccgccg
     1201 aggaacacat ctgcgacttc accaaagaga agtacctgct gggcaagaac gagaaagaat
     1261 actgcgtggt ggaagccaag cccttcgaca gcgtgacctt catctgcccc aagaaaatcg
     1321 gcgcccagtg cttccagaac gtgaacaccg aggacgacat cagcaaagaa aagatggaaa
     1381 gcagcaagct ggccatcgac gagctgctgt acggcagcac cctgtacggc gataccctgc
     1441 tgatctcccc taccgtgaag cagagcaaga ccttctactg cttctgcaac ctcgagatga
     1501 aggacatgaa gaagtatctg cagaagaagc ggctgtctaa agagaaggaa aaagagaaag
     1561 agaacgccaa gaaaaagtcc accgtgaacg tggacaacct ggaaaacgcc gacgaggaca
     1621 tggaagtgat cgtgccccag aagcaggaag atcagaacct cgtccgggcc ctgcacagag
     1681 tgcgcaagat catgaacatc atcgagcgcg agaagaacaa agaggaagaa ggcgacgact
     1741 ccgagaaaaa gaaagaagag gacaaagaag aggaagagga agaagaagaa gaggaagaac
     1801 tggtcgtcga ggaaaaagag gaagaaaagg acaagaacaa gatcgagaag atcatcacca
     1861 aatacggcgt gatgaaggtg gtggtgtcca ccaaccacgc catcaccaag ggctgcgatt
     1921 tcggcaacaa cgtggtcaac tacttcagca agccctaccc catcgagaga tacggcggca
     1981 gcaaagtgtg ccggatcgag gccaagcctg gcgagttcgt gggcttcaag tgcatctacg
     2041 acaacaccgg cagcgtggaa ccccacaact gcttcgacaa ggtgttctac gagggcaaag
     2101 aaaccgacct gcagaccctg atgcccggct acatcagcta cggcaacaag aagaagggca
     2161 aatacgcctt ctacctgaag ctgccccact tcgtgcagca cagctacacc atccggtgca
     2221 agtgccgcag caccgtgtgg cagtacgacg actacgtgtt cgagctggcc gtggaaggcg
     2281 gcgagtctga tgtggtggcc aagagcttcc acgaaggcgc gccgtcgacc tccatcacgg
     2341 cctataagag tgagggggag tcagcggagt tctccttccc actcaacctt ggagaggaaa
     2401 gcctgcaggg agagttgaga tggaaggcag agaaggctcc ttcttcccag tcctggatca
     2461 ccttctccct aaagaaccaa aaggtgtctg tgcagaagtc tactagcaac cccaagttcc
     2521 agctgtccga aacgctccca ctcacccttc agatacccca ggtctccctt cagtttgctg
     2581 gttctggcaa cctgaccctg actctggaca gagggatact gtatcaggaa gtgaacctgg
     2641 tggtgatgaa agtgactcag cccgacagca acactttgac ctgtgaggtg atgggaccca
     2701 cctcacccaa gatgagactg atcttgaagc aggagaatca ggaggccagg gtctccaggc
     2761 aggagaaagt gattcaagtg caggcccctg aagcaggggt gtggcaatgt ctactgagtg
     2821 aaggtgaaga ggtcaagatg gactccaaga tccaggtttt atccaaaggg ttgaattccg
     2881 gcagcctgca ccacatcctg gacgcccaga agatgctgtg gaaccacagg gacaggaacc
     2941 tgcctcctct ggcccctctg ggcccccacc accatcacca ccattgagtc tagacacgtg
     3001 attaattaag gatcccccga cctcgacctc tggctaataa aggaaattta ttttcattgc
     3061 aatagtgtgt tggaattttt tgtgtctctc actcggaagg acatatggga gggcaaatca
     3121 tttggtcgag atccctcgga gatctctagc tagaggatcg atccccgccc cggacgaact
     3181 aaacctgact acgacatctc tgccccttct tcgcggggca gtgcatgtaa tcccttcagt
     3241 tggttggtac aacttgccaa ctgggccctg ttccacatgt gacacggggg gggaccaaac
     3301 acaaaggggt tctctgactg tagttgacat ccttataaat ggatgtgcac atttgccaac
     3361 actgagtggc tttcatcctg gagcagactt tgcagtctgt ggactgcaac acaacattgc
     3421 ctttatgtgt aactcttggc tgaagctctt acaccaatgc tgggggacat gtacctccca
     3481 ggggcccagg aagactacgg gaggctacac caacgtcaat cagaggggcc tgtgtagcta
     3541 ccgataagcg gaccctcaag agggcattag caatagtgtt tataaggccc ccttgttaac
     3601 cctaaacggg tagcatatgc ttcccgggta gtagtatata ctatccagac taaccctaat
     3661 tcaatagcat atgttaccca acgggaagca tatgctatcg aattagggtt agtaaaaggg
     3721 tcctaaggaa cagcgatatc tcccacccca tgagctgtca cggttttatt tacatggggt
     3781 caggattcca cgagggtagt gaaccatttt agtcacaagg gcagtggctg aagatcaagg
     3841 agcgggcagt gaactctcct gaatcttcgc ctgcttcttc attctccttc gtttagctaa
     3901 tagaataact gctgagttgt gaacagtaag gtgtatgtga ggtgctcgaa aacaaggttt
     3961 caggtgacgc ccccagaata aaatttggac ggggggttca gtggtggcat tgtgctatga
     4021 caccaatata accctcacaa accccttggg caataaatac tagtgtagga atgaaacatt
     4081 ctgaatatct ttaacaatag aaatccatgg ggtggggaca agccgtaaag actggatgtc
     4141 catctcacac gaatttatgg ctatgggcaa cacataatcc tagtgcaata tgatactggg
     4201 gttattaaga tgtgtcccag gcagggacca agacaggtga accatgttgt tacactctat
     4261 ttgtaacaag gggaaagaga gtggacgccg acagcagcgg actccactgg ttgtctctaa
     4321 cacccccgaa aattaaacgg ggctccacgc caatggggcc cataaacaaa gacaagtggc
     4381 cactcttttt tttgaaattg tggagtgggg gcacgcgtca gcccccacac gccgccctgc
     4441 ggttttggac tgtaaaataa gggtgtaata acttggctga ttgtaacccc gctaaccact
     4501 gcggtcaaac cacttgccca caaaaccact aatggcaccc cggggaatac ctgcataagt
     4561 aggtgggcgg gccaagatag gggcgcgatt gctgcgatct ggaggacaaa ttacacacac
     4621 ttgcgcctga gcgccaagca cagggttgtt ggtcctcata ttcacgaggt cgctgagagc
     4681 acggtgggct aatgttgcca tgggtagcat atactaccca aatatctgga tagcatatgc
     4741 tatcctaatc tatatctggg tagcataggc tatcctaatc tatatctggg tagcatatgc
     4801 tatcctaatc tatatctggg tagtatatgc tatcctaatt tatatctggg tagcataggc
     4861 tatcctaatc tatatctggg tagcatatgc tatcctaatc tatatctggg tagtatatgc
     4921 tatcctaatc tgtatccggg tagcatatgc tatcctaata gagattaggg tagtatatgc
     4981 tatcctaatt tatatctggg tagcatatac tacccaaata tctggatagc atatgctatc
     5041 ctaatctata tctgggtagc atatgctatc ctaatctata tctgggtagc ataggctatc
     5101 ctaatctata tctgggtagc atatgctatc ctaatctata tctgggtagt atatgctatc
     5161 ctaatttata tctgggtagc ataggctatc ctaatctata tctgggtagc atatgctatc
     5221 ctaatctata tctgggtagt atatgctatc ctaatctgta tccgggtagc atatgctatc
     5281 ctcatgataa gctgtcaaac atgagaattt tcttgaagac gaaagggcct cgtgatacgc
     5341 ctatttttat aggttaatgt catgataata atggtttctt agacgtcagg tggcactttt
     5401 cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat
     5461 ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtatg
     5521 agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt
     5581 tttgctcacc cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcacga
     5641 gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa
     5701 gaacgttttc caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt
     5761 gttgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt
     5821 gagtactcac cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc
     5881 agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac aacgatcgga
     5941 ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat
     6001 cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct
     6061 gcagcaatgg caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc
     6121 cggcaacaat taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg
     6181 gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc
     6241 ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg
     6301 acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca
     6361 ctgattaagc attggtaact gtcagaccaa gtttactcat atatacttta gattgattta
     6421 aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc
     6481 aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa
     6541 ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca
     6601 ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta
     6661 actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc
     6721 caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca
     6781 gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta
     6841 ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag
     6901 cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt
     6961 cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc
     7021 acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac
     7081 ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac
     7141 gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc
     7201 tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgcctttga gtgagctgat
     7261 accgctcgcc gcagccgaac gaccgagcgc agcgagtcag tgagcgagga agcggaagag
     7321 cgcccaatac gcaaaccgcc tctccccgcg cgttggccga ttcattaatg cagctggcac
     7381 gacaggtttc ccgactggaa agcgggcagt gagcgcaacg caattaatgt gagttagctc
     7441 actcattagg caccccaggc tttacacttt atgcttccgg ctcgtatgtt gtgtggaatt
     7501 gtgagcggat aacaatttca cacaggaaac agctatgacc atgattacgc caagctctag
     7561 ctagaggtcg accaattctc atgtttgaca gcttatcatc gcagatccgg gcaacgttgt
     7621 tgccattgct gcaggcgcag aactggtagg tatggaagat ctatacattg aatcaatatt
     7681 ggcaattagc catattagtc attggttata tagcataaat caatattggc tattggccat
     7741 tgcatacgtt gtatctatat cataatatgt acatttatat tggctcatgt ccaatatgac
     7801 cgc
//
