LOCUS       sequence-245752-        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        1..380
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        381..584
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     535..555
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     581..605
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     primer_bind     1086..1105
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     regulatory      1128..1137
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3885..3902
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    3954..4009
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(4008..4027)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(6245..6263)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6331..6435
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6436..6504,6505..7296)
                     /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(6654..6673)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7467..8055
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7956..7975
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     8209..8226
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    8343..8364
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(8379..8384,8385..8402,8403..8409)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8417..8433
                     /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     8422..8444
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     8441..8457
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     8441..8457
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc
       61 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
      121 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
      181 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
      241 aagtgtatca tatgccaagt ccgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
      301 ggcattatgc ccagtacatg accttacggg actttcctac ttggcagtac atctacgtat
      361 tagtcatcgc tattaccatg gtgatgcggt tttggcagta caccaatggg cgtggatagc
      421 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt
      481 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaataa ccccgccccg ttgacgcaaa
      541 tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctcgttta gtgaaccgtc
      601 agatcctcac tctcttccgc atcgctgtct gcgagggcca gctgttgggc tcgcggttga
      661 ggacaaactc ttcgcggtct ttccagtact cttggatcgg aaacccgtcg gcctccgaac
      721 ggtactccgc caccgaggga cctgagcgag tccgcatcga ccggatcgga aaacctctcg
      781 agaaaggcgt ctaaccagtc acagtcgcaa ggtaggctga gcaccgtggc gggcggcagc
      841 gggtggcggt cggggttgtt tctggcggag gtgctgctga tgatgtaatt aaagtaggcg
      901 gtcttgagac ggcggatggt cgaggtgagg tgtggcaggc ttgagatcca gctgttgggg
      961 tgagtactcc ctctcaaaag cgggcattac ttctgcgcta agattgtcag tttccaaaaa
     1021 cgaggaggat ttgatattca cctggcccga tctggccata cacttgagtg acaatgacat
     1081 ccactttgcc tttctctcca caggtgtcca ctcccaggtc caagtttgcc accatggagt
     1141 ttcagaccca ggtactcatg tccctgctgc tctgcatgtc tggtgcggcc gccctgatca
     1201 gaaacggcaa caatcctcag gccctggtgc ccgacaacag cggcgatcct aatgccggcc
     1261 agaacaaggc cagcagcgac aaccaggata cctgcgagat ccagaaaatg gccgaagaga
     1321 tgatggaaaa gatgatgaaa gaaaaggacg tgttcagcag catcatggaa cccctgcaga
     1381 gcaagctgac cgacgaccgg ctgtgcagca agatgaagta caccaacatc tgcctgcacg
     1441 agaaggacaa gacccctctg accttcccct gcaccaaccc tcagtacgag cagctgatcc
     1501 agcagttcac ctacaagaag ctgtgcaaca gcaaggtggc cttcagcaac gtgctgctca
     1561 agagcttcat cgacaagaag aacgaggaaa acaccttcaa cgccatcatc cagaactaca
     1621 aggtgctgag cacatgcatc gacgaggacc tgaaggacat ctacaacgcc gccatcgagc
     1681 tgttcagcga cctgagaacc agcgtgcgcg agatcaccga gaaactgtgg tccaagaaca
     1741 tgatcgaggt gctgaaaacc agagagcagg ccattgccgg catcctgtgc gagctgcgga
     1801 atggcaacaa tgccgccctg gtgtccaaca gcctgagcta cgagaacttt ggcatcctga
     1861 aagtgaacta cgagggcctg atcaaccagg cctacaaggc cttctccgac tactacagct
     1921 acttccccgc cttcgccatc cggctgctgg aaaaggatgg cctggtggaa cggctggtgg
     1981 ccatccacga gagcctgacc aactaccgga cccggaacat cctgaagaag atcaacgaga
     2041 agtccaagaa tgaggtcctg aacaacgagg acatcatgca cagcctgtcc agctacaagc
     2101 accacgctgg cggcaccaga ggcagcttca tgcagtatag aggcgtgggc cagctgagca
     2161 agggcggcct gtctgtggat gagaagggcg gacagcagat cgccagcgcc gtgggaaatc
     2221 aggccgccaa tatggtggcc cctgccccta aggacagcag ccctacaatg gccgctccca
     2281 gcaccgacac cgtgaccaac aacatggcca caaccaccac cgcccctgcc accgctaata
     2341 tggctgcccc tagcaccaat accgccaccg ccaacatggc agctccatcc accaacacag
     2401 ccacagcaaa tatggccgca ccttccaccg atacagtgac taccaatatg gctaccacca
     2461 ccacagtgcc cgccacagct aataccgccg ccagccccga taccaacacc gacagctcca
     2521 cctaccctct gtacggcacc gagagcagca agaccaagga cgtggtggtg ctcgtgcggg
     2581 acctgctgaa ggataccaat atcatcaagt tcgagaagaa tgagcccacc agccagatcg
     2641 acgacgaggg cattaagaag ctgatcgagt cctccttctt cgacctgtcc gacaacacca
     2701 tgctgatgcg gctgatcatc aagccccagg ccagcatcct gtttatcatc cagtccttta
     2761 tcatgatgac ccctagcccc acccgggacg cccggatgta ctgcaagaag aagctggtca
     2821 acggacagct catcgagaac aacgatctga aggccgaaac cgaagaggaa gatatgatca
     2881 acgaattcag ctccaagtac aacctgttct acgagcggct gaagatggaa gaactgagag
     2941 agatcgagca ggaccggaag tccctgaaga actccaaggg caacctggcc gtgctggaag
     3001 tgcggaacag ccagaacggc cccgacggca aagaagtgaa cggcgcaggc gacgccgcca
     3061 acggaaacaa catgaacggc ggcaacaacg gcgctgccag cagcctgatc gtggtcgtca
     3121 gagatgacct ggccgagaaa accgacgaca tcatcaagaa caatgtggac ctggaaagcc
     3181 tgaaagccga cgtggaacag gccttccgga acttcgagta ccagagcggc tctggcgcgc
     3241 cgtcgacctc catcacggcc tataagagtg agggggagtc agcggagttc tccttcccac
     3301 tcaaccttgg agaggaaagc ctgcagggag agttgagatg gaaggcagag aaggctcctt
     3361 cttcccagtc ctggatcacc ttctccctaa agaaccaaaa ggtgtctgtg cagaagtcta
     3421 ctagcaaccc caagttccag ctgtccgaaa cgctcccact cacccttcag ataccccagg
     3481 tctcccttca gtttgctggt tctggcaacc tgaccctgac tctggacaga gggatactgt
     3541 atcaggaagt gaacctggtg gtgatgaaag tgactcagcc cgacagcaac actttgacct
     3601 gtgaggtgat gggacccacc tcacccaaga tgagactgat cttgaagcag gagaatcagg
     3661 aggccagggt ctccaggcag gagaaagtga ttcaagtgca ggcccctgaa gcaggggtgt
     3721 ggcaatgtct actgagtgaa ggtgaagagg tcaagatgga ctccaagatc caggttttat
     3781 ccaaagggtt gaattccggc agcctgcacc acatcctgga cgcccagaag atgctgtgga
     3841 accacaggga caggaacctg cctcctctgg cccctctggg cccccaccac catcaccacc
     3901 attgagtcta gacacgtgat taattaagga tcccccgacc tcgacctctg gctaataaag
     3961 gaaatttatt ttcattgcaa tagtgtgttg gaattttttg tgtctctcac tcggaaggac
     4021 atatgggagg gcaaatcatt tggtcgagat ccctcggaga tctctagcta gaggatcgat
     4081 ccccgccccg gacgaactaa acctgactac gacatctctg ccccttcttc gcggggcagt
     4141 gcatgtaatc ccttcagttg gttggtacaa cttgccaact gggccctgtt ccacatgtga
     4201 cacggggggg gaccaaacac aaaggggttc tctgactgta gttgacatcc ttataaatgg
     4261 atgtgcacat ttgccaacac tgagtggctt tcatcctgga gcagactttg cagtctgtgg
     4321 actgcaacac aacattgcct ttatgtgtaa ctcttggctg aagctcttac accaatgctg
     4381 ggggacatgt acctcccagg ggcccaggaa gactacggga ggctacacca acgtcaatca
     4441 gaggggcctg tgtagctacc gataagcgga ccctcaagag ggcattagca atagtgttta
     4501 taaggccccc ttgttaaccc taaacgggta gcatatgctt cccgggtagt agtatatact
     4561 atccagacta accctaattc aatagcatat gttacccaac gggaagcata tgctatcgaa
     4621 ttagggttag taaaagggtc ctaaggaaca gcgatatctc ccaccccatg agctgtcacg
     4681 gttttattta catggggtca ggattccacg agggtagtga accattttag tcacaagggc
     4741 agtggctgaa gatcaaggag cgggcagtga actctcctga atcttcgcct gcttcttcat
     4801 tctccttcgt ttagctaata gaataactgc tgagttgtga acagtaaggt gtatgtgagg
     4861 tgctcgaaaa caaggtttca ggtgacgccc ccagaataaa atttggacgg ggggttcagt
     4921 ggtggcattg tgctatgaca ccaatataac cctcacaaac cccttgggca ataaatacta
     4981 gtgtaggaat gaaacattct gaatatcttt aacaatagaa atccatgggg tggggacaag
     5041 ccgtaaagac tggatgtcca tctcacacga atttatggct atgggcaaca cataatccta
     5101 gtgcaatatg atactggggt tattaagatg tgtcccaggc agggaccaag acaggtgaac
     5161 catgttgtta cactctattt gtaacaaggg gaaagagagt ggacgccgac agcagcggac
     5221 tccactggtt gtctctaaca cccccgaaaa ttaaacgggg ctccacgcca atggggccca
     5281 taaacaaaga caagtggcca ctcttttttt tgaaattgtg gagtgggggc acgcgtcagc
     5341 ccccacacgc cgccctgcgg ttttggactg taaaataagg gtgtaataac ttggctgatt
     5401 gtaaccccgc taaccactgc ggtcaaacca cttgcccaca aaaccactaa tggcaccccg
     5461 gggaatacct gcataagtag gtgggcgggc caagataggg gcgcgattgc tgcgatctgg
     5521 aggacaaatt acacacactt gcgcctgagc gccaagcaca gggttgttgg tcctcatatt
     5581 cacgaggtcg ctgagagcac ggtgggctaa tgttgccatg ggtagcatat actacccaaa
     5641 tatctggata gcatatgcta tcctaatcta tatctgggta gcataggcta tcctaatcta
     5701 tatctgggta gcatatgcta tcctaatcta tatctgggta gtatatgcta tcctaattta
     5761 tatctgggta gcataggcta tcctaatcta tatctgggta gcatatgcta tcctaatcta
     5821 tatctgggta gtatatgcta tcctaatctg tatccgggta gcatatgcta tcctaataga
     5881 gattagggta gtatatgcta tcctaattta tatctgggta gcatatacta cccaaatatc
     5941 tggatagcat atgctatcct aatctatatc tgggtagcat atgctatcct aatctatatc
     6001 tgggtagcat aggctatcct aatctatatc tgggtagcat atgctatcct aatctatatc
     6061 tgggtagtat atgctatcct aatttatatc tgggtagcat aggctatcct aatctatatc
     6121 tgggtagcat atgctatcct aatctatatc tgggtagtat atgctatcct aatctgtatc
     6181 cgggtagcat atgctatcct catgataagc tgtcaaacat gagaattttc ttgaagacga
     6241 aagggcctcg tgatacgcct atttttatag gttaatgtca tgataataat ggtttcttag
     6301 acgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa
     6361 atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat
     6421 tgaaaaagga agagtatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg
     6481 gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa
     6541 gatcagttgg gtgcacgagt gggttacatc gaactggatc tcaacagcgg taagatcctt
     6601 gagagttttc gccccgaaga acgttttcca atgatgagca cttttaaagt tctgctatgt
     6661 ggcgcggtat tatcccgtgt tgacgccggg caagagcaac tcggtcgccg catacactat
     6721 tctcagaatg acttggttga gtactcacca gtcacagaaa agcatcttac ggatggcatg
     6781 acagtaagag aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta
     6841 cttctgacaa cgatcggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat
     6901 catgtaactc gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag
     6961 cgtgacacca cgatgcctgc agcaatggca acaacgttgc gcaaactatt aactggcgaa
     7021 ctacttactc tagcttcccg gcaacaatta atagactgga tggaggcgga taaagttgca
     7081 ggaccacttc tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc
     7141 ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt
     7201 atcgtagtta tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc
     7261 gctgagatag gtgcctcact gattaagcat tggtaactgt cagaccaagt ttactcatat
     7321 atactttaga ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt
     7381 tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac
     7441 cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc
     7501 ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca
     7561 actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta
     7621 gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct
     7681 ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg
     7741 gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc
     7801 acacagccca gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta
     7861 tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg
     7921 gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt
     7981 cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg
     8041 cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg
     8101 ccttttgctc acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc
     8161 gcctttgagt gagctgatac cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg
     8221 agcgaggaag cggaagagcg cccaatacgc aaaccgcctc tccccgcgcg ttggccgatt
     8281 cattaatgca gctggcacga caggtttccc gactggaaag cgggcagtga gcgcaacgca
     8341 attaatgtga gttagctcac tcattaggca ccccaggctt tacactttat gcttccggct
     8401 cgtatgttgt gtggaattgt gagcggataa caatttcaca caggaaacag ctatgaccat
     8461 gattacgcca agctctagct agaggtcgac caattctcat gtttgacagc ttatcatcgc
     8521 agatccgggc aacgttgttg ccattgctgc aggcgcagaa ctggtaggta tggaagatct
     8581 atacattgaa tcaatattgg caattagcca tattagtcat tggttatata gcataaatca
     8641 atattggcta ttggccattg catacgttgt atctatatca taatatgtac atttatattg
     8701 gctcatgtcc aatatgaccg ccatgtt
//
