LOCUS       sequence-242834-        7686 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 7686)
  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 7686)
  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_MSP5-Cd4-bio-His
FEATURES             Location/Qualifiers
     source          1..7686
                     /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             2850..2867
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    2919..2974
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(2973..2992)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(5210..5228)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5296..5400
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5401..5469,5470..6261)
                     /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(5619..5638)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6432..7020
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6921..6940
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7174..7191
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    7308..7329
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7344..7349,7350..7367,7368..7374)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7382..7398
                     /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     7387..7409
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7406..7422
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7406..7422
                     /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 gcggccgccc
     1201 atcctctgaa gttcagcttc tggcgggaag agaagatcca cctccagatc tacaccaacc
     1261 ggctgctgag agaggaaggc aagaacgaca acgtggaaca gatggaaagc cccagcatca
     1321 gcggcaccga gggcaagaaa gaaatccaga tgatcagcca tctgcagctc cagagcggca
     1381 agcacgacca ggtgtccttt ctgagcgcca acgacgccaa cctgaagggc gccaatgagg
     1441 aagcctctgc cgagagcggc gagaacggca agaagtccga cgaggaaaac gtgaagaaaa
     1501 gcgacgaaga gaacgccaaa aagtctgatg aagagaacaa ggacgccaac agcaacacca
     1561 aggatgccga gtccgccgag ggcgaagaga atcccgtgtc ccaggaaaac cagatgaaga
     1621 ccctgaacaa cgaagaggac aagaccaacg atggcaagaa gaacggggac gaggaagata
     1681 agaagggcga aaagggcaag gacgacggca agggcgagga aggctccgag gaaaagatgg
     1741 aagagtccgc caagaaagag gaagagaata ccgaggccaa caagaacctg gaaagcaagc
     1801 acgccgaggc cgaggtgtcc gagcacaagc aggatgtgac aacccctgag ggcggcaagg
     1861 atagccccga aggcgagaac cagaaagaga acacccctaa caacgccaac caggaaaatc
     1921 aggaaaacgc cgacaacgac gagaacaagc tgcacctgga caagctggac gacgacgtgc
     1981 cacactacag cgccctgcgg aacaaccggg tggacaaggg catcaccgac agcatgatgc
     2041 tgaagaacat catcgaggat aacaccaaga gctgcagcgt gaacaacggc ggctgcgccg
     2101 acgaccagat ctgcatccgg atcaacaaca tgggcatcaa gtgcatctgc aaagagggcc
     2161 acctgttcgg cggcaagtgt atcctgagca gaagctctgg cgcgccgtcg acctccatca
     2221 cggcctataa gagtgagggg gagtcagcgg agttctcctt cccactcaac cttggagagg
     2281 aaagcctgca gggagagttg agatggaagg cagagaaggc tccttcttcc cagtcctgga
     2341 tcaccttctc cctaaagaac caaaaggtgt ctgtgcagaa gtctactagc aaccccaagt
     2401 tccagctgtc cgaaacgctc ccactcaccc ttcagatacc ccaggtctcc cttcagtttg
     2461 ctggttctgg caacctgacc ctgactctgg acagagggat actgtatcag gaagtgaacc
     2521 tggtggtgat gaaagtgact cagcccgaca gcaacacttt gacctgtgag gtgatgggac
     2581 ccacctcacc caagatgaga ctgatcttga agcaggagaa tcaggaggcc agggtctcca
     2641 ggcaggagaa agtgattcaa gtgcaggccc ctgaagcagg ggtgtggcaa tgtctactga
     2701 gtgaaggtga agaggtcaag atggactcca agatccaggt tttatccaaa gggttgaatt
     2761 ccggcagcct gcaccacatc ctggacgccc agaagatgct gtggaaccac agggacagga
     2821 acctgcctcc tctggcccct ctgggccccc accaccatca ccaccattga gtctagacac
     2881 gtgattaatt aaggatcccc cgacctcgac ctctggctaa taaaggaaat ttattttcat
     2941 tgcaatagtg tgttggaatt ttttgtgtct ctcactcgga aggacatatg ggagggcaaa
     3001 tcatttggtc gagatccctc ggagatctct agctagagga tcgatccccg ccccggacga
     3061 actaaacctg actacgacat ctctgcccct tcttcgcggg gcagtgcatg taatcccttc
     3121 agttggttgg tacaacttgc caactgggcc ctgttccaca tgtgacacgg ggggggacca
     3181 aacacaaagg ggttctctga ctgtagttga catccttata aatggatgtg cacatttgcc
     3241 aacactgagt ggctttcatc ctggagcaga ctttgcagtc tgtggactgc aacacaacat
     3301 tgcctttatg tgtaactctt ggctgaagct cttacaccaa tgctggggga catgtacctc
     3361 ccaggggccc aggaagacta cgggaggcta caccaacgtc aatcagaggg gcctgtgtag
     3421 ctaccgataa gcggaccctc aagagggcat tagcaatagt gtttataagg cccccttgtt
     3481 aaccctaaac gggtagcata tgcttcccgg gtagtagtat atactatcca gactaaccct
     3541 aattcaatag catatgttac ccaacgggaa gcatatgcta tcgaattagg gttagtaaaa
     3601 gggtcctaag gaacagcgat atctcccacc ccatgagctg tcacggtttt atttacatgg
     3661 ggtcaggatt ccacgagggt agtgaaccat tttagtcaca agggcagtgg ctgaagatca
     3721 aggagcgggc agtgaactct cctgaatctt cgcctgcttc ttcattctcc ttcgtttagc
     3781 taatagaata actgctgagt tgtgaacagt aaggtgtatg tgaggtgctc gaaaacaagg
     3841 tttcaggtga cgcccccaga ataaaatttg gacggggggt tcagtggtgg cattgtgcta
     3901 tgacaccaat ataaccctca caaacccctt gggcaataaa tactagtgta ggaatgaaac
     3961 attctgaata tctttaacaa tagaaatcca tggggtgggg acaagccgta aagactggat
     4021 gtccatctca cacgaattta tggctatggg caacacataa tcctagtgca atatgatact
     4081 ggggttatta agatgtgtcc caggcaggga ccaagacagg tgaaccatgt tgttacactc
     4141 tatttgtaac aaggggaaag agagtggacg ccgacagcag cggactccac tggttgtctc
     4201 taacaccccc gaaaattaaa cggggctcca cgccaatggg gcccataaac aaagacaagt
     4261 ggccactctt ttttttgaaa ttgtggagtg ggggcacgcg tcagccccca cacgccgccc
     4321 tgcggttttg gactgtaaaa taagggtgta ataacttggc tgattgtaac cccgctaacc
     4381 actgcggtca aaccacttgc ccacaaaacc actaatggca ccccggggaa tacctgcata
     4441 agtaggtggg cgggccaaga taggggcgcg attgctgcga tctggaggac aaattacaca
     4501 cacttgcgcc tgagcgccaa gcacagggtt gttggtcctc atattcacga ggtcgctgag
     4561 agcacggtgg gctaatgttg ccatgggtag catatactac ccaaatatct ggatagcata
     4621 tgctatccta atctatatct gggtagcata ggctatccta atctatatct gggtagcata
     4681 tgctatccta atctatatct gggtagtata tgctatccta atttatatct gggtagcata
     4741 ggctatccta atctatatct gggtagcata tgctatccta atctatatct gggtagtata
     4801 tgctatccta atctgtatcc gggtagcata tgctatccta atagagatta gggtagtata
     4861 tgctatccta atttatatct gggtagcata tactacccaa atatctggat agcatatgct
     4921 atcctaatct atatctgggt agcatatgct atcctaatct atatctgggt agcataggct
     4981 atcctaatct atatctgggt agcatatgct atcctaatct atatctgggt agtatatgct
     5041 atcctaattt atatctgggt agcataggct atcctaatct atatctgggt agcatatgct
     5101 atcctaatct atatctgggt agtatatgct atcctaatct gtatccgggt agcatatgct
     5161 atcctcatga taagctgtca aacatgagaa ttttcttgaa gacgaaaggg cctcgtgata
     5221 cgcctatttt tataggttaa tgtcatgata ataatggttt cttagacgtc aggtggcact
     5281 tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg
     5341 tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt
     5401 atgagtattc aacatttccg tgtcgccctt attccctttt ttgcggcatt ttgccttcct
     5461 gtttttgctc acccagaaac gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca
     5521 cgagtgggtt acatcgaact ggatctcaac agcggtaaga tccttgagag ttttcgcccc
     5581 gaagaacgtt ttccaatgat gagcactttt aaagttctgc tatgtggcgc ggtattatcc
     5641 cgtgttgacg ccgggcaaga gcaactcggt cgccgcatac actattctca gaatgacttg
     5701 gttgagtact caccagtcac agaaaagcat cttacggatg gcatgacagt aagagaatta
     5761 tgcagtgctg ccataaccat gagtgataac actgcggcca acttacttct gacaacgatc
     5821 ggaggaccga aggagctaac cgcttttttg cacaacatgg gggatcatgt aactcgcctt
     5881 gatcgttggg aaccggagct gaatgaagcc ataccaaacg acgagcgtga caccacgatg
     5941 cctgcagcaa tggcaacaac gttgcgcaaa ctattaactg gcgaactact tactctagct
     6001 tcccggcaac aattaataga ctggatggag gcggataaag ttgcaggacc acttctgcgc
     6061 tcggcccttc cggctggctg gtttattgct gataaatctg gagccggtga gcgtgggtct
     6121 cgcggtatca ttgcagcact ggggccagat ggtaagccct cccgtatcgt agttatctac
     6181 acgacgggga gtcaggcaac tatggatgaa cgaaatagac agatcgctga gataggtgcc
     6241 tcactgatta agcattggta actgtcagac caagtttact catatatact ttagattgat
     6301 ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga taatctcatg
     6361 accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc
     6421 aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa
     6481 ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag
     6541 gtaactggct tcagcagagc gcagatacca aatactgttc ttctagtgta gccgtagtta
     6601 ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta
     6661 ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag
     6721 ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg
     6781 gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg
     6841 cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag
     6901 cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc
     6961 cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa
     7021 aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg
     7081 ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgcctt tgagtgagct
     7141 gataccgctc gccgcagccg aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa
     7201 gagcgcccaa tacgcaaacc gcctctcccc gcgcgttggc cgattcatta atgcagctgg
     7261 cacgacaggt ttcccgactg gaaagcgggc agtgagcgca acgcaattaa tgtgagttag
     7321 ctcactcatt aggcacccca ggctttacac tttatgcttc cggctcgtat gttgtgtgga
     7381 attgtgagcg gataacaatt tcacacagga aacagctatg accatgatta cgccaagctc
     7441 tagctagagg tcgaccaatt ctcatgtttg acagcttatc atcgcagatc cgggcaacgt
     7501 tgttgccatt gctgcaggcg cagaactggt aggtatggaa gatctataca ttgaatcaat
     7561 attggcaatt agccatatta gtcattggtt atatagcata aatcaatatt ggctattggc
     7621 cattgcatac gttgtatcta tatcataata tgtacattta tattggctca tgtccaatat
     7681 gaccgc
//
