LOCUS       sequence-245739-        7221 bp    DNA     circular SYN 15-APR-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 7221)
  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 7221)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 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_MSP4-Cd4-bio-His
FEATURES             Location/Qualifiers
     source          1..7221
                     /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             2379..2396
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    2448..2503
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(2502..2521)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(4739..4757)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4825..4929
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4930..4998,4999..5790)
                     /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(5148..5167)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5961..6549
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6450..6469
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6703..6720
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    6837..6858
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6873..6878,6879..6896,6897..6903)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6911..6927
                     /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     6916..6938
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     6935..6951
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6935..6951
                     /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 gcctacaacc
     1201 tgaagatcat caccaacaag atcatctaca agcgggtgct gaccaacaac aacgacaaca
     1261 atgccttcac caaagagaca ggccagagca gcagcaccaa gggcaccagc ggcgacagca
     1321 acaagcagaa catcgacgac agccctcaga agggcctgag cgaggacggc aagtctgtgt
     1381 ccgccggcag ccccatcacc agcaagtaca gcgagcagtc cggcgacaac atcaacctgg
     1441 ccaacacccc taacagcgag gacagcggca atgccggcac cttcagcggc aactccaata
     1501 gcgccggcat cgtggaaggc agcgacgaca acgatggcga cgaggacgag gatgaagacg
     1561 aagatgagga cgaggacgaa gataagaacc tgtgcctgca caacaacggc ggctgcggcg
     1621 aggacaagct gtgcgagtac ctgggccgga agatcgtgaa gtgcttctgc aaagagggct
     1681 acaagctcgt cggcaccgac tgcgtgaagt ccagcgagag cagctctggc gcgccgtcga
     1741 cctccatcac ggcctataag agtgaggggg agtcagcgga gttctccttc ccactcaacc
     1801 ttggagagga aagcctgcag ggagagttga gatggaaggc agagaaggct ccttcttccc
     1861 agtcctggat caccttctcc ctaaagaacc aaaaggtgtc tgtgcagaag tctactagca
     1921 accccaagtt ccagctgtcc gaaacgctcc cactcaccct tcagataccc caggtctccc
     1981 ttcagtttgc tggttctggc aacctgaccc tgactctgga cagagggata ctgtatcagg
     2041 aagtgaacct ggtggtgatg aaagtgactc agcccgacag caacactttg acctgtgagg
     2101 tgatgggacc cacctcaccc aagatgagac tgatcttgaa gcaggagaat caggaggcca
     2161 gggtctccag gcaggagaaa gtgattcaag tgcaggcccc tgaagcaggg gtgtggcaat
     2221 gtctactgag tgaaggtgaa gaggtcaaga tggactccaa gatccaggtt ttatccaaag
     2281 ggttgaattc cggcagcctg caccacatcc tggacgccca gaagatgctg tggaaccaca
     2341 gggacaggaa cctgcctcct ctggcccctc tgggccccca ccaccatcac caccattgag
     2401 tctagacacg tgattaatta aggatccccc gacctcgacc tctggctaat aaaggaaatt
     2461 tattttcatt gcaatagtgt gttggaattt tttgtgtctc tcactcggaa ggacatatgg
     2521 gagggcaaat catttggtcg agatccctcg gagatctcta gctagaggat cgatccccgc
     2581 cccggacgaa ctaaacctga ctacgacatc tctgcccctt cttcgcgggg cagtgcatgt
     2641 aatcccttca gttggttggt acaacttgcc aactgggccc tgttccacat gtgacacggg
     2701 gggggaccaa acacaaaggg gttctctgac tgtagttgac atccttataa atggatgtgc
     2761 acatttgcca acactgagtg gctttcatcc tggagcagac tttgcagtct gtggactgca
     2821 acacaacatt gcctttatgt gtaactcttg gctgaagctc ttacaccaat gctgggggac
     2881 atgtacctcc caggggccca ggaagactac gggaggctac accaacgtca atcagagggg
     2941 cctgtgtagc taccgataag cggaccctca agagggcatt agcaatagtg tttataaggc
     3001 ccccttgtta accctaaacg ggtagcatat gcttcccggg tagtagtata tactatccag
     3061 actaacccta attcaatagc atatgttacc caacgggaag catatgctat cgaattaggg
     3121 ttagtaaaag ggtcctaagg aacagcgata tctcccaccc catgagctgt cacggtttta
     3181 tttacatggg gtcaggattc cacgagggta gtgaaccatt ttagtcacaa gggcagtggc
     3241 tgaagatcaa ggagcgggca gtgaactctc ctgaatcttc gcctgcttct tcattctcct
     3301 tcgtttagct aatagaataa ctgctgagtt gtgaacagta aggtgtatgt gaggtgctcg
     3361 aaaacaaggt ttcaggtgac gcccccagaa taaaatttgg acggggggtt cagtggtggc
     3421 attgtgctat gacaccaata taaccctcac aaaccccttg ggcaataaat actagtgtag
     3481 gaatgaaaca ttctgaatat ctttaacaat agaaatccat ggggtgggga caagccgtaa
     3541 agactggatg tccatctcac acgaatttat ggctatgggc aacacataat cctagtgcaa
     3601 tatgatactg gggttattaa gatgtgtccc aggcagggac caagacaggt gaaccatgtt
     3661 gttacactct atttgtaaca aggggaaaga gagtggacgc cgacagcagc ggactccact
     3721 ggttgtctct aacacccccg aaaattaaac ggggctccac gccaatgggg cccataaaca
     3781 aagacaagtg gccactcttt tttttgaaat tgtggagtgg gggcacgcgt cagcccccac
     3841 acgccgccct gcggttttgg actgtaaaat aagggtgtaa taacttggct gattgtaacc
     3901 ccgctaacca ctgcggtcaa accacttgcc cacaaaacca ctaatggcac cccggggaat
     3961 acctgcataa gtaggtgggc gggccaagat aggggcgcga ttgctgcgat ctggaggaca
     4021 aattacacac acttgcgcct gagcgccaag cacagggttg ttggtcctca tattcacgag
     4081 gtcgctgaga gcacggtggg ctaatgttgc catgggtagc atatactacc caaatatctg
     4141 gatagcatat gctatcctaa tctatatctg ggtagcatag gctatcctaa tctatatctg
     4201 ggtagcatat gctatcctaa tctatatctg ggtagtatat gctatcctaa tttatatctg
     4261 ggtagcatag gctatcctaa tctatatctg ggtagcatat gctatcctaa tctatatctg
     4321 ggtagtatat gctatcctaa tctgtatccg ggtagcatat gctatcctaa tagagattag
     4381 ggtagtatat gctatcctaa tttatatctg ggtagcatat actacccaaa tatctggata
     4441 gcatatgcta tcctaatcta tatctgggta gcatatgcta tcctaatcta tatctgggta
     4501 gcataggcta tcctaatcta tatctgggta gcatatgcta tcctaatcta tatctgggta
     4561 gtatatgcta tcctaattta tatctgggta gcataggcta tcctaatcta tatctgggta
     4621 gcatatgcta tcctaatcta tatctgggta gtatatgcta tcctaatctg tatccgggta
     4681 gcatatgcta tcctcatgat aagctgtcaa acatgagaat tttcttgaag acgaaagggc
     4741 ctcgtgatac gcctattttt ataggttaat gtcatgataa taatggtttc ttagacgtca
     4801 ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat
     4861 tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa
     4921 aggaagagta tgagtattca acatttccgt gtcgccctta ttcccttttt tgcggcattt
     4981 tgccttcctg tttttgctca cccagaaacg ctggtgaaag taaaagatgc tgaagatcag
     5041 ttgggtgcac gagtgggtta catcgaactg gatctcaaca gcggtaagat ccttgagagt
     5101 tttcgccccg aagaacgttt tccaatgatg agcactttta aagttctgct atgtggcgcg
     5161 gtattatccc gtgttgacgc cgggcaagag caactcggtc gccgcataca ctattctcag
     5221 aatgacttgg ttgagtactc accagtcaca gaaaagcatc ttacggatgg catgacagta
     5281 agagaattat gcagtgctgc cataaccatg agtgataaca ctgcggccaa cttacttctg
     5341 acaacgatcg gaggaccgaa ggagctaacc gcttttttgc acaacatggg ggatcatgta
     5401 actcgccttg atcgttggga accggagctg aatgaagcca taccaaacga cgagcgtgac
     5461 accacgatgc ctgcagcaat ggcaacaacg ttgcgcaaac tattaactgg cgaactactt
     5521 actctagctt cccggcaaca attaatagac tggatggagg cggataaagt tgcaggacca
     5581 cttctgcgct cggcccttcc ggctggctgg tttattgctg ataaatctgg agccggtgag
     5641 cgtgggtctc gcggtatcat tgcagcactg gggccagatg gtaagccctc ccgtatcgta
     5701 gttatctaca cgacggggag tcaggcaact atggatgaac gaaatagaca gatcgctgag
     5761 ataggtgcct cactgattaa gcattggtaa ctgtcagacc aagtttactc atatatactt
     5821 tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat
     5881 aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta
     5941 gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa
     6001 acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt
     6061 tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct tctagtgtag
     6121 ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta
     6181 atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca
     6241 agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag
     6301 cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa
     6361 agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga
     6421 acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc
     6481 gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc
     6541 ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt
     6601 gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat taccgccttt
     6661 gagtgagctg ataccgctcg ccgcagccga acgaccgagc gcagcgagtc agtgagcgag
     6721 gaagcggaag agcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa
     6781 tgcagctggc acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat
     6841 gtgagttagc tcactcatta ggcaccccag gctttacact ttatgcttcc ggctcgtatg
     6901 ttgtgtggaa ttgtgagcgg ataacaattt cacacaggaa acagctatga ccatgattac
     6961 gccaagctct agctagaggt cgaccaattc tcatgtttga cagcttatca tcgcagatcc
     7021 gggcaacgtt gttgccattg ctgcaggcgc agaactggta ggtatggaag atctatacat
     7081 tgaatcaata ttggcaatta gccatattag tcattggtta tatagcataa atcaatattg
     7141 gctattggcc attgcatacg ttgtatctat atcataatat gtacatttat attggctcat
     7201 gtccaatatg accgccatgt t
//
