LOCUS       sequence-242832-        8727 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 8727)
  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 8727)
  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_GAMA-Cd4-bio-His
FEATURES             Location/Qualifiers
     source          1..8727
                     /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             3891..3908
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    3960..4015
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(4014..4033)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(6251..6269)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6337..6441
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6442..6510,6511..7302)
                     /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(6660..6679)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7473..8061
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7962..7981
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     8215..8232
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    8349..8370
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(8385..8390,8391..8408,8409..8415)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8423..8439
                     /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     8428..8450
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     8447..8463
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     8447..8463
                     /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 tgatcagaaa cggcaacaat cctcaggccc tggtgcccga caacagcggc gatcctaatg
     1261 ccggccagaa caaggccagc agcgacaacc aggatacctg cgagatccag aaaatggccg
     1321 aagagatgat ggaaaagatg atgaaagaaa aggacgtgtt cagcagcatc atggaacccc
     1381 tgcagagcaa gctgaccgac gaccggctgt gcagcaagat gaagtacacc aacatctgcc
     1441 tgcacgagaa ggacaagacc cctctgacct tcccctgcac caaccctcag tacgagcagc
     1501 tgatccagca gttcacctac aagaagctgt gcaacagcaa ggtggccttc agcaacgtgc
     1561 tgctcaagag cttcatcgac aagaagaacg aggaaaacac cttcaacgcc atcatccaga
     1621 actacaaggt gctgagcaca tgcatcgacg aggacctgaa ggacatctac aacgccgcca
     1681 tcgagctgtt cagcgacctg agaaccagcg tgcgcgagat caccgagaaa ctgtggtcca
     1741 agaacatgat cgaggtgctg aaaaccagag agcaggccat tgccggcatc ctgtgcgagc
     1801 tgcggaatgg caacaatgcc gccctggtgt ccaacagcct gagctacgag aactttggca
     1861 tcctgaaagt gaactacgag ggcctgatca accaggccta caaggccttc tccgactact
     1921 acagctactt ccccgccttc gccatccggc tgctggaaaa ggatggcctg gtggaacggc
     1981 tggtggccat ccacgagagc ctgaccaact accggacccg gaacatcctg aagaagatca
     2041 acgagaagtc caagaatgag gtcctgaaca acgaggacat catgcacagc ctgtccagct
     2101 acaagcacca cgctggcggc accagaggca gcttcatgca gtatagaggc gtgggccagc
     2161 tgagcaaggg cggcctgtct gtggatgaga agggcggaca gcagatcgcc agcgccgtgg
     2221 gaaatcaggc cgccaatatg gtggcccctg cccctaagga cagcagccct acaatggccg
     2281 ctcccagcac cgacaccgtg accaacaaca tggccacaac caccaccgcc cctgccaccg
     2341 ctaatatggc tgcccctagc accaataccg ccaccgccaa catggcagct ccatccacca
     2401 acacagccac agcaaatatg gccgcacctt ccaccgatac agtgactacc aatatggcta
     2461 ccaccaccac agtgcccgcc acagctaata ccgccgccag ccccgatacc aacaccgaca
     2521 gctccaccta ccctctgtac ggcaccgaga gcagcaagac caaggacgtg gtggtgctcg
     2581 tgcgggacct gctgaaggat accaatatca tcaagttcga gaagaatgag cccaccagcc
     2641 agatcgacga cgagggcatt aagaagctga tcgagtcctc cttcttcgac ctgtccgaca
     2701 acaccatgct gatgcggctg atcatcaagc cccaggccag catcctgttt atcatccagt
     2761 cctttatcat gatgacccct agccccaccc gggacgcccg gatgtactgc aagaagaagc
     2821 tggtcaacgg acagctcatc gagaacaacg atctgaaggc cgaaaccgaa gaggaagata
     2881 tgatcaacga attcagctcc aagtacaacc tgttctacga gcggctgaag atggaagaac
     2941 tgagagagat cgagcaggac cggaagtccc tgaagaactc caagggcaac ctggccgtgc
     3001 tggaagtgcg gaacagccag aacggccccg acggcaaaga agtgaacggc gcaggcgacg
     3061 ccgccaacgg aaacaacatg aacggcggca acaacggcgc tgccagcagc ctgatcgtgg
     3121 tcgtcagaga tgacctggcc gagaaaaccg acgacatcat caagaacaat gtggacctgg
     3181 aaagcctgaa agccgacgtg gaacaggcct tccggaactt cgagtaccag agcggctctg
     3241 gcgcgccgtc gacctccatc acggcctata agagtgaggg ggagtcagcg gagttctcct
     3301 tcccactcaa ccttggagag gaaagcctgc agggagagtt gagatggaag gcagagaagg
     3361 ctccttcttc ccagtcctgg atcaccttct ccctaaagaa ccaaaaggtg tctgtgcaga
     3421 agtctactag caaccccaag ttccagctgt ccgaaacgct cccactcacc cttcagatac
     3481 cccaggtctc ccttcagttt gctggttctg gcaacctgac cctgactctg gacagaggga
     3541 tactgtatca ggaagtgaac ctggtggtga tgaaagtgac tcagcccgac agcaacactt
     3601 tgacctgtga ggtgatggga cccacctcac ccaagatgag actgatcttg aagcaggaga
     3661 atcaggaggc cagggtctcc aggcaggaga aagtgattca agtgcaggcc cctgaagcag
     3721 gggtgtggca atgtctactg agtgaaggtg aagaggtcaa gatggactcc aagatccagg
     3781 ttttatccaa agggttgaat tccggcagcc tgcaccacat cctggacgcc cagaagatgc
     3841 tgtggaacca cagggacagg aacctgcctc ctctggcccc tctgggcccc caccaccatc
     3901 accaccattg agtctagaca cgtgattaat taaggatccc ccgacctcga cctctggcta
     3961 ataaaggaaa tttattttca ttgcaatagt gtgttggaat tttttgtgtc tctcactcgg
     4021 aaggacatat gggagggcaa atcatttggt cgagatccct cggagatctc tagctagagg
     4081 atcgatcccc gccccggacg aactaaacct gactacgaca tctctgcccc ttcttcgcgg
     4141 ggcagtgcat gtaatccctt cagttggttg gtacaacttg ccaactgggc cctgttccac
     4201 atgtgacacg gggggggacc aaacacaaag gggttctctg actgtagttg acatccttat
     4261 aaatggatgt gcacatttgc caacactgag tggctttcat cctggagcag actttgcagt
     4321 ctgtggactg caacacaaca ttgcctttat gtgtaactct tggctgaagc tcttacacca
     4381 atgctggggg acatgtacct cccaggggcc caggaagact acgggaggct acaccaacgt
     4441 caatcagagg ggcctgtgta gctaccgata agcggaccct caagagggca ttagcaatag
     4501 tgtttataag gcccccttgt taaccctaaa cgggtagcat atgcttcccg ggtagtagta
     4561 tatactatcc agactaaccc taattcaata gcatatgtta cccaacggga agcatatgct
     4621 atcgaattag ggttagtaaa agggtcctaa ggaacagcga tatctcccac cccatgagct
     4681 gtcacggttt tatttacatg gggtcaggat tccacgaggg tagtgaacca ttttagtcac
     4741 aagggcagtg gctgaagatc aaggagcggg cagtgaactc tcctgaatct tcgcctgctt
     4801 cttcattctc cttcgtttag ctaatagaat aactgctgag ttgtgaacag taaggtgtat
     4861 gtgaggtgct cgaaaacaag gtttcaggtg acgcccccag aataaaattt ggacgggggg
     4921 ttcagtggtg gcattgtgct atgacaccaa tataaccctc acaaacccct tgggcaataa
     4981 atactagtgt aggaatgaaa cattctgaat atctttaaca atagaaatcc atggggtggg
     5041 gacaagccgt aaagactgga tgtccatctc acacgaattt atggctatgg gcaacacata
     5101 atcctagtgc aatatgatac tggggttatt aagatgtgtc ccaggcaggg accaagacag
     5161 gtgaaccatg ttgttacact ctatttgtaa caaggggaaa gagagtggac gccgacagca
     5221 gcggactcca ctggttgtct ctaacacccc cgaaaattaa acggggctcc acgccaatgg
     5281 ggcccataaa caaagacaag tggccactct tttttttgaa attgtggagt gggggcacgc
     5341 gtcagccccc acacgccgcc ctgcggtttt ggactgtaaa ataagggtgt aataacttgg
     5401 ctgattgtaa ccccgctaac cactgcggtc aaaccacttg cccacaaaac cactaatggc
     5461 accccgggga atacctgcat aagtaggtgg gcgggccaag ataggggcgc gattgctgcg
     5521 atctggagga caaattacac acacttgcgc ctgagcgcca agcacagggt tgttggtcct
     5581 catattcacg aggtcgctga gagcacggtg ggctaatgtt gccatgggta gcatatacta
     5641 cccaaatatc tggatagcat atgctatcct aatctatatc tgggtagcat aggctatcct
     5701 aatctatatc tgggtagcat atgctatcct aatctatatc tgggtagtat atgctatcct
     5761 aatttatatc tgggtagcat aggctatcct aatctatatc tgggtagcat atgctatcct
     5821 aatctatatc tgggtagtat atgctatcct aatctgtatc cgggtagcat atgctatcct
     5881 aatagagatt agggtagtat atgctatcct aatttatatc tgggtagcat atactaccca
     5941 aatatctgga tagcatatgc tatcctaatc tatatctggg tagcatatgc tatcctaatc
     6001 tatatctggg tagcataggc tatcctaatc tatatctggg tagcatatgc tatcctaatc
     6061 tatatctggg tagtatatgc tatcctaatt tatatctggg tagcataggc tatcctaatc
     6121 tatatctggg tagcatatgc tatcctaatc tatatctggg tagtatatgc tatcctaatc
     6181 tgtatccggg tagcatatgc tatcctcatg ataagctgtc aaacatgaga attttcttga
     6241 agacgaaagg gcctcgtgat acgcctattt ttataggtta atgtcatgat aataatggtt
     6301 tcttagacgt caggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt
     6361 ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa
     6421 taatattgaa aaaggaagag tatgagtatt caacatttcc gtgtcgccct tattcccttt
     6481 tttgcggcat tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat
     6541 gctgaagatc agttgggtgc acgagtgggt tacatcgaac tggatctcaa cagcggtaag
     6601 atccttgaga gttttcgccc cgaagaacgt tttccaatga tgagcacttt taaagttctg
     6661 ctatgtggcg cggtattatc ccgtgttgac gccgggcaag agcaactcgg tcgccgcata
     6721 cactattctc agaatgactt ggttgagtac tcaccagtca cagaaaagca tcttacggat
     6781 ggcatgacag taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc
     6841 aacttacttc tgacaacgat cggaggaccg aaggagctaa ccgctttttt gcacaacatg
     6901 ggggatcatg taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac
     6961 gacgagcgtg acaccacgat gcctgcagca atggcaacaa cgttgcgcaa actattaact
     7021 ggcgaactac ttactctagc ttcccggcaa caattaatag actggatgga ggcggataaa
     7081 gttgcaggac cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct
     7141 ggagccggtg agcgtgggtc tcgcggtatc attgcagcac tggggccaga tggtaagccc
     7201 tcccgtatcg tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga
     7261 cagatcgctg agataggtgc ctcactgatt aagcattggt aactgtcaga ccaagtttac
     7321 tcatatatac tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag
     7381 atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg
     7441 tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc
     7501 tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag
     7561 ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtt
     7621 cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac
     7681 ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc
     7741 gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt
     7801 tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt
     7861 gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc
     7921 ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt
     7981 tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca
     8041 ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt
     8101 tgctggcctt ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt
     8161 attaccgcct ttgagtgagc tgataccgct cgccgcagcc gaacgaccga gcgcagcgag
     8221 tcagtgagcg aggaagcgga agagcgccca atacgcaaac cgcctctccc cgcgcgttgg
     8281 ccgattcatt aatgcagctg gcacgacagg tttcccgact ggaaagcggg cagtgagcgc
     8341 aacgcaatta atgtgagtta gctcactcat taggcacccc aggctttaca ctttatgctt
     8401 ccggctcgta tgttgtgtgg aattgtgagc ggataacaat ttcacacagg aaacagctat
     8461 gaccatgatt acgccaagct ctagctagag gtcgaccaat tctcatgttt gacagcttat
     8521 catcgcagat ccgggcaacg ttgttgccat tgctgcaggc gcagaactgg taggtatgga
     8581 agatctatac attgaatcaa tattggcaat tagccatatt agtcattggt tatatagcat
     8641 aaatcaatat tggctattgg ccattgcata cgttgtatct atatcataat atgtacattt
     8701 atattggctc atgtccaata tgaccgc
//
