LOCUS       sequence-245946-        7782 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 7782)
  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 7782)
  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_P38-Cd4-bio-His
FEATURES             Location/Qualifiers
     source          1..7782
                     /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             2946..2963
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    3015..3070
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(3069..3088)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(5306..5324)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5392..5496
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5497..5565,5566..6357)
                     /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(5715..5734)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6528..7116
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7017..7036
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7270..7287
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    7404..7425
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7440..7445,7446..7463,7464..7470)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7478..7494
                     /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     7483..7505
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7502..7518
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7502..7518
                     /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 gcggccgcca
     1201 aacaccacgt ggacctgagc aatagcgcca agagcacctt cctgctgaac gtggtgcccg
     1261 gcgacaccgt ggtgtacaca tgcccctaca gcttcaacaa ggacatgaag cacatctgcg
     1321 ccagagagcg ggacttcttc gaccggaagc tgttctgctt cgaccacgtg atcatcaacc
     1381 ggaacgtgtt ccagctgcgg gactacgtgc ggggagccta caacatcgtg tccaaatacg
     1441 tgaacaacgt gtacacctcc gagttcaccg tgcctcccgt ggtgctgatg aaccggcact
     1501 tcgagtgcta ctgctacctg gacgacggcg gcagaatcca gaaaaagacc ctgaaggtgc
     1561 acatcagcaa gggcatcgtg aagcggaccc ctggctgcga cttcaacgac gagtacagag
     1621 agagcaccgc catcaccacc ttcaacaaca tgaaccggaa gatcgccaag gcctgcgaca
     1681 gctaccctag aggcggcgat accatcaccc tgctgtgccc cgtgaactac gccgtgcagc
     1741 ccgacggctg cttcaaccag gtgtacgtga agaaagagga catccagaac gacaagaaca
     1801 agctcgagga acggttcaat atcgcccgga agtgggagca ggacaagtac aaggtggtgt
     1861 ccatcgagac actgttcggc aagaaactgg acagcgtggg cgacgagatc accagattcg
     1921 ccaagatccc cgtgaccaac gatgagatca gcttcacctg tacctgcaag gccaacgacg
     1981 gcagcgacac cctgatgatg aatgtgtaca tcaacgaggc ctacgacaag ttcgtgcaga
     2041 gcagccacga caagagcgac cagtccaacc agggcaaccc cgtgcagtct aatcagggaa
     2101 atccagtgca cagcaaccag gggaatcctg tccagagtaa tcaggggaac cccatccaga
     2161 gcaactacaa caagtacatc gagagcaaca agcgcttcgt gtaccccaac gatggccggg
     2221 tcgagtacac ccggcacaag ttcagcagca cctccaccgt gtcccggacc gaggaaagaa
     2281 gcggagccag ctctggcgcg ccgtcgacct ccatcacggc ctataagagt gagggggagt
     2341 cagcggagtt ctccttccca ctcaaccttg gagaggaaag cctgcaggga gagttgagat
     2401 ggaaggcaga gaaggctcct tcttcccagt cctggatcac cttctcccta aagaaccaaa
     2461 aggtgtctgt gcagaagtct actagcaacc ccaagttcca gctgtccgaa acgctcccac
     2521 tcacccttca gataccccag gtctcccttc agtttgctgg ttctggcaac ctgaccctga
     2581 ctctggacag agggatactg tatcaggaag tgaacctggt ggtgatgaaa gtgactcagc
     2641 ccgacagcaa cactttgacc tgtgaggtga tgggacccac ctcacccaag atgagactga
     2701 tcttgaagca ggagaatcag gaggccaggg tctccaggca ggagaaagtg attcaagtgc
     2761 aggcccctga agcaggggtg tggcaatgtc tactgagtga aggtgaagag gtcaagatgg
     2821 actccaagat ccaggtttta tccaaagggt tgaattccgg cagcctgcac cacatcctgg
     2881 acgcccagaa gatgctgtgg aaccacaggg acaggaacct gcctcctctg gcccctctgg
     2941 gcccccacca ccatcaccac cattgagtct agacacgtga ttaattaagg atcccccgac
     3001 ctcgacctct ggctaataaa ggaaatttat tttcattgca atagtgtgtt ggaatttttt
     3061 gtgtctctca ctcggaagga catatgggag ggcaaatcat ttggtcgaga tccctcggag
     3121 atctctagct agaggatcga tccccgcccc ggacgaacta aacctgacta cgacatctct
     3181 gccccttctt cgcggggcag tgcatgtaat cccttcagtt ggttggtaca acttgccaac
     3241 tgggccctgt tccacatgtg acacgggggg ggaccaaaca caaaggggtt ctctgactgt
     3301 agttgacatc cttataaatg gatgtgcaca tttgccaaca ctgagtggct ttcatcctgg
     3361 agcagacttt gcagtctgtg gactgcaaca caacattgcc tttatgtgta actcttggct
     3421 gaagctctta caccaatgct gggggacatg tacctcccag gggcccagga agactacggg
     3481 aggctacacc aacgtcaatc agaggggcct gtgtagctac cgataagcgg accctcaaga
     3541 gggcattagc aatagtgttt ataaggcccc cttgttaacc ctaaacgggt agcatatgct
     3601 tcccgggtag tagtatatac tatccagact aaccctaatt caatagcata tgttacccaa
     3661 cgggaagcat atgctatcga attagggtta gtaaaagggt cctaaggaac agcgatatct
     3721 cccaccccat gagctgtcac ggttttattt acatggggtc aggattccac gagggtagtg
     3781 aaccatttta gtcacaaggg cagtggctga agatcaagga gcgggcagtg aactctcctg
     3841 aatcttcgcc tgcttcttca ttctccttcg tttagctaat agaataactg ctgagttgtg
     3901 aacagtaagg tgtatgtgag gtgctcgaaa acaaggtttc aggtgacgcc cccagaataa
     3961 aatttggacg gggggttcag tggtggcatt gtgctatgac accaatataa ccctcacaaa
     4021 ccccttgggc aataaatact agtgtaggaa tgaaacattc tgaatatctt taacaataga
     4081 aatccatggg gtggggacaa gccgtaaaga ctggatgtcc atctcacacg aatttatggc
     4141 tatgggcaac acataatcct agtgcaatat gatactgggg ttattaagat gtgtcccagg
     4201 cagggaccaa gacaggtgaa ccatgttgtt acactctatt tgtaacaagg ggaaagagag
     4261 tggacgccga cagcagcgga ctccactggt tgtctctaac acccccgaaa attaaacggg
     4321 gctccacgcc aatggggccc ataaacaaag acaagtggcc actctttttt ttgaaattgt
     4381 ggagtggggg cacgcgtcag cccccacacg ccgccctgcg gttttggact gtaaaataag
     4441 ggtgtaataa cttggctgat tgtaaccccg ctaaccactg cggtcaaacc acttgcccac
     4501 aaaaccacta atggcacccc ggggaatacc tgcataagta ggtgggcggg ccaagatagg
     4561 ggcgcgattg ctgcgatctg gaggacaaat tacacacact tgcgcctgag cgccaagcac
     4621 agggttgttg gtcctcatat tcacgaggtc gctgagagca cggtgggcta atgttgccat
     4681 gggtagcata tactacccaa atatctggat agcatatgct atcctaatct atatctgggt
     4741 agcataggct atcctaatct atatctgggt agcatatgct atcctaatct atatctgggt
     4801 agtatatgct atcctaattt atatctgggt agcataggct atcctaatct atatctgggt
     4861 agcatatgct atcctaatct atatctgggt agtatatgct atcctaatct gtatccgggt
     4921 agcatatgct atcctaatag agattagggt agtatatgct atcctaattt atatctgggt
     4981 agcatatact acccaaatat ctggatagca tatgctatcc taatctatat ctgggtagca
     5041 tatgctatcc taatctatat ctgggtagca taggctatcc taatctatat ctgggtagca
     5101 tatgctatcc taatctatat ctgggtagta tatgctatcc taatttatat ctgggtagca
     5161 taggctatcc taatctatat ctgggtagca tatgctatcc taatctatat ctgggtagta
     5221 tatgctatcc taatctgtat ccgggtagca tatgctatcc tcatgataag ctgtcaaaca
     5281 tgagaatttt cttgaagacg aaagggcctc gtgatacgcc tatttttata ggttaatgtc
     5341 atgataataa tggtttctta gacgtcaggt ggcacttttc ggggaaatgt gcgcggaacc
     5401 cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc
     5461 tgataaatgc ttcaataata ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc
     5521 gcccttattc ccttttttgc ggcattttgc cttcctgttt ttgctcaccc agaaacgctg
     5581 gtgaaagtaa aagatgctga agatcagttg ggtgcacgag tgggttacat cgaactggat
     5641 ctcaacagcg gtaagatcct tgagagtttt cgccccgaag aacgttttcc aatgatgagc
     5701 acttttaaag ttctgctatg tggcgcggta ttatcccgtg ttgacgccgg gcaagagcaa
     5761 ctcggtcgcc gcatacacta ttctcagaat gacttggttg agtactcacc agtcacagaa
     5821 aagcatctta cggatggcat gacagtaaga gaattatgca gtgctgccat aaccatgagt
     5881 gataacactg cggccaactt acttctgaca acgatcggag gaccgaagga gctaaccgct
     5941 tttttgcaca acatggggga tcatgtaact cgccttgatc gttgggaacc ggagctgaat
     6001 gaagccatac caaacgacga gcgtgacacc acgatgcctg cagcaatggc aacaacgttg
     6061 cgcaaactat taactggcga actacttact ctagcttccc ggcaacaatt aatagactgg
     6121 atggaggcgg ataaagttgc aggaccactt ctgcgctcgg cccttccggc tggctggttt
     6181 attgctgata aatctggagc cggtgagcgt gggtctcgcg gtatcattgc agcactgggg
     6241 ccagatggta agccctcccg tatcgtagtt atctacacga cggggagtca ggcaactatg
     6301 gatgaacgaa atagacagat cgctgagata ggtgcctcac tgattaagca ttggtaactg
     6361 tcagaccaag tttactcata tatactttag attgatttaa aacttcattt ttaatttaaa
     6421 aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt
     6481 tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt
     6541 tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt
     6601 ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag
     6661 ataccaaata ctgttcttct agtgtagccg tagttaggcc accacttcaa gaactctgta
     6721 gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat
     6781 aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg
     6841 ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg
     6901 agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac
     6961 aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga
     7021 aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt
     7081 ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta
     7141 cggttcctgg ccttttgctg gccttttgct cacatgttct ttcctgcgtt atcccctgat
     7201 tctgtggata accgtattac cgcctttgag tgagctgata ccgctcgccg cagccgaacg
     7261 accgagcgca gcgagtcagt gagcgaggaa gcggaagagc gcccaatacg caaaccgcct
     7321 ctccccgcgc gttggccgat tcattaatgc agctggcacg acaggtttcc cgactggaaa
     7381 gcgggcagtg agcgcaacgc aattaatgtg agttagctca ctcattaggc accccaggct
     7441 ttacacttta tgcttccggc tcgtatgttg tgtggaattg tgagcggata acaatttcac
     7501 acaggaaaca gctatgacca tgattacgcc aagctctagc tagaggtcga ccaattctca
     7561 tgtttgacag cttatcatcg cagatccggg caacgttgtt gccattgctg caggcgcaga
     7621 actggtaggt atggaagatc tatacattga atcaatattg gcaattagcc atattagtca
     7681 ttggttatat agcataaatc aatattggct attggccatt gcatacgttg tatctatatc
     7741 ataatatgta catttatatt ggctcatgtc caatatgacc gc
//
