LOCUS       sequence-245651-        8916 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of recombinant P. malariae protein in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8916)
  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 8916)
  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: Pm_GAMA-Cd4-bio-His
FEATURES             Location/Qualifiers
     source          1..8916
                     /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             4074..4091
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    4143..4198
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(4197..4216)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(6434..6452)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6520..6624
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6625..6693,6694..7485)
                     /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(6843..6862)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7656..8244
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8145..8164
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     8398..8415
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    8532..8553
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(8568..8573,8574..8591,8592..8598)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8606..8622
                     /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     8611..8633
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     8630..8646
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     8630..8646
                     /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 caacaacgcc caggccctga ttcccgacaa gaacaatgcc aagaacgaca
     1261 accagaacag catgagtaac aacaacaaca acaacgagaa tacctgcgag atccagaaca
     1321 tggccgagga aatgatggaa aagctgctga acgagaagga cgtgttcagc tccatcatgg
     1381 aagccctgca gagcaagctg agcgacgaca gcctgtgcac ccagaccaag tacaccaaca
     1441 tctgcatcaa cgaaaaggac aaggtgcccc tgagcttccc ctgcaccagc cagaagtacg
     1501 agcacctgat ccaccagttc acctacaaga agctgtgcca cagcaagatc gccttcagca
     1561 acatcctgct gaagtccttc atcgacaaaa acaacgaaga gaacaccttc aacaatatca
     1621 tcaagaacta caatgtgctg ctgagctgca tcgataacga cctgaaggac atctacacca
     1681 gcagcatcaa gctgttcagc gacctgcggt ccagcatcgt ggacgtgacc gagaaactgt
     1741 ggtccaagaa tatcgccgac gtgctgaaga agagagagca tctgatcgcc ggcatcttct
     1801 gcggcctgag gaacggcgcc aagagcaacc tggtgtccaa cagcctgagc tacgagaacc
     1861 tgggcatcct gaagatcaac agcgaggaac tgatcaacga ggccttctac gccttctccg
     1921 agtactacta caacttccct ctgttcacca tgcagctgct ggaaaggggc ggactggtgg
     1981 aaaggctggt ggccatccac gagaagctga ccaactaccg gacccggaat atcctgaaca
     2041 agatcaatga gcggagcaag aatgaggtgc tgaacaatga ggatgtcctg cacaacctgg
     2101 ccaactacaa gcaccactcc cagaaaacaa atggcgcctt cgtgcagtcc aaagagatca
     2161 agatcatcag caacgtgggc ccctacaaca atgcaaacga gaacgccaac cctgccgcca
     2221 acgccgccac aaatgccgcc accaataccg acgccaatgc taacaccaac gccaatagta
     2281 atgccaacac ctacgtggac gccaacacaa ataccaacgc taacaaaaac gacaatctga
     2341 acgcaaacac caattccaac ctgaatgcca ataccaatag caacctcaac gccaacaaga
     2401 acgtgaacac aaacgccaac actaacgcta acgctaatac caacactaat gccaactcca
     2461 acgctaattc taacgccaac agcaacagca actccaatgc aaacagcaat gctaataccg
     2521 ccactaactc tgatgcaaac acaaacgcaa ataccaatgc aaacacaaat gccaacacaa
     2581 atgctaatac caacgcaaat accgccacca acctggacgc caatactgcc atcaacagcc
     2641 agggcgggct gcacgccaac gaccctctga acaaccctct gtacagcacc tccagcctga
     2701 agcccaagaa tgtggcccag ctgatcaagg acctgattaa gaatctggac atcgtgaagt
     2761 tcgaaaacaa tgaacccacc aacaaaatcg acgaggaagg catcaaaaag ctgatcggca
     2821 acagcttcct ggacctgtcc gacaacatca tgctcgtgcg gctgctcatc aagccccagg
     2881 ccgccatcct gttcatcatc cagagcttcg tgatgatgac ccctagcccc acccgggacg
     2941 ctaagacctt ctgcaagaag gccctggtca acggacagct gattgacagc accaatctgg
     3001 gcgtggaatc cgaggaagag aatctcatca acaacttcgc cagcaagtac aacctgatct
     3061 acgaaaagat caagctcgaa gaactgcgcg agctggaaca gagcaaaaag accctgaagt
     3121 ctagcaatag ccacctgagc gcactgcaag tccgaaacac cgagaacaac aaagagtcca
     3181 acacagagag caacaccgaa agcaatactg agagtaatac cgagtctaac aagcagaaca
     3241 acaaccaggg catgaacttc aacaacaatt tcaatctcaa caacgccgct cccctgatcg
     3301 ctatcgtgtc cgatcctctg ggcgagaagg ccgatgacat cattaagaac aacctcgacc
     3361 tgagcagcct gtccagcggc gtcgagcaga ccttccagac actgaacgcc aattccagat
     3421 ctggcgcgcc gtcgacctcc atcacggcct ataagagtga gggggagtca gcggagttct
     3481 ccttcccact caaccttgga gaggaaagcc tgcagggaga gttgagatgg aaggcagaga
     3541 aggctccttc ttcccagtcc tggatcacct tctccctaaa gaaccaaaag gtgtctgtgc
     3601 agaagtctac tagcaacccc aagttccagc tgtccgaaac gctcccactc acccttcaga
     3661 taccccaggt ctcccttcag tttgctggtt ctggcaacct gaccctgact ctggacagag
     3721 ggatactgta tcaggaagtg aacctggtgg tgatgaaagt gactcagccc gacagcaaca
     3781 ctttgacctg tgaggtgatg ggacccacct cacccaagat gagactgatc ttgaagcagg
     3841 agaatcagga ggccagggtc tccaggcagg agaaagtgat tcaagtgcag gcccctgaag
     3901 caggggtgtg gcaatgtcta ctgagtgaag gtgaagaggt caagatggac tccaagatcc
     3961 aggttttatc caaagggttg aattccggca gcctgcacca catcctggac gcccagaaga
     4021 tgctgtggaa ccacagggac aggaacctgc ctcctctggc ccctctgggc ccccaccacc
     4081 atcaccacca ttgagtctag acacgtgatt aattaaggat cccccgacct cgacctctgg
     4141 ctaataaagg aaatttattt tcattgcaat agtgtgttgg aattttttgt gtctctcact
     4201 cggaaggaca tatgggaggg caaatcattt ggtcgagatc cctcggagat ctctagctag
     4261 aggatcgatc cccgccccgg acgaactaaa cctgactacg acatctctgc cccttcttcg
     4321 cggggcagtg catgtaatcc cttcagttgg ttggtacaac ttgccaactg ggccctgttc
     4381 cacatgtgac acgggggggg accaaacaca aaggggttct ctgactgtag ttgacatcct
     4441 tataaatgga tgtgcacatt tgccaacact gagtggcttt catcctggag cagactttgc
     4501 agtctgtgga ctgcaacaca acattgcctt tatgtgtaac tcttggctga agctcttaca
     4561 ccaatgctgg gggacatgta cctcccaggg gcccaggaag actacgggag gctacaccaa
     4621 cgtcaatcag aggggcctgt gtagctaccg ataagcggac cctcaagagg gcattagcaa
     4681 tagtgtttat aaggccccct tgttaaccct aaacgggtag catatgcttc ccgggtagta
     4741 gtatatacta tccagactaa ccctaattca atagcatatg ttacccaacg ggaagcatat
     4801 gctatcgaat tagggttagt aaaagggtcc taaggaacag cgatatctcc caccccatga
     4861 gctgtcacgg ttttatttac atggggtcag gattccacga gggtagtgaa ccattttagt
     4921 cacaagggca gtggctgaag atcaaggagc gggcagtgaa ctctcctgaa tcttcgcctg
     4981 cttcttcatt ctccttcgtt tagctaatag aataactgct gagttgtgaa cagtaaggtg
     5041 tatgtgaggt gctcgaaaac aaggtttcag gtgacgcccc cagaataaaa tttggacggg
     5101 gggttcagtg gtggcattgt gctatgacac caatataacc ctcacaaacc ccttgggcaa
     5161 taaatactag tgtaggaatg aaacattctg aatatcttta acaatagaaa tccatggggt
     5221 ggggacaagc cgtaaagact ggatgtccat ctcacacgaa tttatggcta tgggcaacac
     5281 ataatcctag tgcaatatga tactggggtt attaagatgt gtcccaggca gggaccaaga
     5341 caggtgaacc atgttgttac actctatttg taacaagggg aaagagagtg gacgccgaca
     5401 gcagcggact ccactggttg tctctaacac ccccgaaaat taaacggggc tccacgccaa
     5461 tggggcccat aaacaaagac aagtggccac tctttttttt gaaattgtgg agtgggggca
     5521 cgcgtcagcc cccacacgcc gccctgcggt tttggactgt aaaataaggg tgtaataact
     5581 tggctgattg taaccccgct aaccactgcg gtcaaaccac ttgcccacaa aaccactaat
     5641 ggcaccccgg ggaatacctg cataagtagg tgggcgggcc aagatagggg cgcgattgct
     5701 gcgatctgga ggacaaatta cacacacttg cgcctgagcg ccaagcacag ggttgttggt
     5761 cctcatattc acgaggtcgc tgagagcacg gtgggctaat gttgccatgg gtagcatata
     5821 ctacccaaat atctggatag catatgctat cctaatctat atctgggtag cataggctat
     5881 cctaatctat atctgggtag catatgctat cctaatctat atctgggtag tatatgctat
     5941 cctaatttat atctgggtag cataggctat cctaatctat atctgggtag catatgctat
     6001 cctaatctat atctgggtag tatatgctat cctaatctgt atccgggtag catatgctat
     6061 cctaatagag attagggtag tatatgctat cctaatttat atctgggtag catatactac
     6121 ccaaatatct ggatagcata tgctatccta atctatatct gggtagcata tgctatccta
     6181 atctatatct gggtagcata ggctatccta atctatatct gggtagcata tgctatccta
     6241 atctatatct gggtagtata tgctatccta atttatatct gggtagcata ggctatccta
     6301 atctatatct gggtagcata tgctatccta atctatatct gggtagtata tgctatccta
     6361 atctgtatcc gggtagcata tgctatcctc atgataagct gtcaaacatg agaattttct
     6421 tgaagacgaa agggcctcgt gatacgccta tttttatagg ttaatgtcat gataataatg
     6481 gtttcttaga cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta
     6541 tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt
     6601 caataatatt gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc ccttattccc
     6661 ttttttgcgg cattttgcct tcctgttttt gctcacccag aaacgctggt gaaagtaaaa
     6721 gatgctgaag atcagttggg tgcacgagtg ggttacatcg aactggatct caacagcggt
     6781 aagatccttg agagttttcg ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt
     6841 ctgctatgtg gcgcggtatt atcccgtgtt gacgccgggc aagagcaact cggtcgccgc
     6901 atacactatt ctcagaatga cttggttgag tactcaccag tcacagaaaa gcatcttacg
     6961 gatggcatga cagtaagaga attatgcagt gctgccataa ccatgagtga taacactgcg
     7021 gccaacttac ttctgacaac gatcggagga ccgaaggagc taaccgcttt tttgcacaac
     7081 atgggggatc atgtaactcg ccttgatcgt tgggaaccgg agctgaatga agccatacca
     7141 aacgacgagc gtgacaccac gatgcctgca gcaatggcaa caacgttgcg caaactatta
     7201 actggcgaac tacttactct agcttcccgg caacaattaa tagactggat ggaggcggat
     7261 aaagttgcag gaccacttct gcgctcggcc cttccggctg gctggtttat tgctgataaa
     7321 tctggagccg gtgagcgtgg gtctcgcggt atcattgcag cactggggcc agatggtaag
     7381 ccctcccgta tcgtagttat ctacacgacg gggagtcagg caactatgga tgaacgaaat
     7441 agacagatcg ctgagatagg tgcctcactg attaagcatt ggtaactgtc agaccaagtt
     7501 tactcatata tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg
     7561 aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga
     7621 gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta
     7681 atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa
     7741 gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact
     7801 gttcttctag tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca
     7861 tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt
     7921 accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg
     7981 ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag
     8041 cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta
     8101 agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat
     8161 ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg
     8221 tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc
     8281 ttttgctggc cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac
     8341 cgtattaccg cctttgagtg agctgatacc gctcgccgca gccgaacgac cgagcgcagc
     8401 gagtcagtga gcgaggaagc ggaagagcgc ccaatacgca aaccgcctct ccccgcgcgt
     8461 tggccgattc attaatgcag ctggcacgac aggtttcccg actggaaagc gggcagtgag
     8521 cgcaacgcaa ttaatgtgag ttagctcact cattaggcac cccaggcttt acactttatg
     8581 cttccggctc gtatgttgtg tggaattgtg agcggataac aatttcacac aggaaacagc
     8641 tatgaccatg attacgccaa gctctagcta gaggtcgacc aattctcatg tttgacagct
     8701 tatcatcgca gatccgggca acgttgttgc cattgctgca ggcgcagaac tggtaggtat
     8761 ggaagatcta tacattgaat caatattggc aattagccat attagtcatt ggttatatag
     8821 cataaatcaa tattggctat tggccattgc atacgttgta tctatatcat aatatgtaca
     8881 tttatattgg ctcatgtcca atatgaccgc catgtt
//
