LOCUS       sequence-245799-        7158 bp    DNA     circular SYN 15-APR-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 7158)
  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 7158)
  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: Pk_MSP4-Cd4-bio-His
FEATURES             Location/Qualifiers
     source          1..7158
                     /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             2322..2339
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    2391..2446
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(2445..2464)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(4682..4700)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4768..4872
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4873..4941,4942..5733)
                     /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(5091..5110)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5904..6492
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6393..6412
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6646..6663
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    6780..6801
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6816..6821,6822..6839,6840..6846)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6854..6870
                     /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     6859..6881
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     6878..6894
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6878..6894
                     /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 gcggccgccg
     1201 atttcagatc catcgtgggc cggatcagac acggccggat tctggaagat gccggcgata
     1261 agggcagcgg cgccaacgac gccaacaacg gcgacaccag agacaacagc agacctcccg
     1321 ccgagaacaa tcacgccgag ggcggcgata gcaccacccc tcccaatgat gcccagaaag
     1381 agggcgccca gaacatcctg cctggcaatg ctggcgaggc cggcaaccac aacgtggaag
     1441 aacccaacga ggactacgac gcccacgacg acgagtacta caacaatgtg gaagatgacg
     1501 agagctacga cctgaaggac cacaacgaga acctgtgctc tgagaacaat ggcggctgcg
     1561 gcaacgacaa gatctgcgag aacatcggcg acggcatcgt gaagtgcctg tgcaagcccg
     1621 gctacaagct cgtgggcacc gagtgcgtgg aagccagcaa gagcagctct ggcgcgccgt
     1681 cgacctccat cacggcctat aagagtgagg gggagtcagc ggagttctcc ttcccactca
     1741 accttggaga ggaaagcctg cagggagagt tgagatggaa ggcagagaag gctccttctt
     1801 cccagtcctg gatcaccttc tccctaaaga accaaaaggt gtctgtgcag aagtctacta
     1861 gcaaccccaa gttccagctg tccgaaacgc tcccactcac ccttcagata ccccaggtct
     1921 cccttcagtt tgctggttct ggcaacctga ccctgactct ggacagaggg atactgtatc
     1981 aggaagtgaa cctggtggtg atgaaagtga ctcagcccga cagcaacact ttgacctgtg
     2041 aggtgatggg acccacctca cccaagatga gactgatctt gaagcaggag aatcaggagg
     2101 ccagggtctc caggcaggag aaagtgattc aagtgcaggc ccctgaagca ggggtgtggc
     2161 aatgtctact gagtgaaggt gaagaggtca agatggactc caagatccag gttttatcca
     2221 aagggttgaa ttccggcagc ctgcaccaca tcctggacgc ccagaagatg ctgtggaacc
     2281 acagggacag gaacctgcct cctctggccc ctctgggccc ccaccaccat caccaccatt
     2341 gagtctagac acgtgattaa ttaaggatcc cccgacctcg acctctggct aataaaggaa
     2401 atttattttc attgcaatag tgtgttggaa ttttttgtgt ctctcactcg gaaggacata
     2461 tgggagggca aatcatttgg tcgagatccc tcggagatct ctagctagag gatcgatccc
     2521 cgccccggac gaactaaacc tgactacgac atctctgccc cttcttcgcg gggcagtgca
     2581 tgtaatccct tcagttggtt ggtacaactt gccaactggg ccctgttcca catgtgacac
     2641 ggggggggac caaacacaaa ggggttctct gactgtagtt gacatcctta taaatggatg
     2701 tgcacatttg ccaacactga gtggctttca tcctggagca gactttgcag tctgtggact
     2761 gcaacacaac attgccttta tgtgtaactc ttggctgaag ctcttacacc aatgctgggg
     2821 gacatgtacc tcccaggggc ccaggaagac tacgggaggc tacaccaacg tcaatcagag
     2881 gggcctgtgt agctaccgat aagcggaccc tcaagagggc attagcaata gtgtttataa
     2941 ggcccccttg ttaaccctaa acgggtagca tatgcttccc gggtagtagt atatactatc
     3001 cagactaacc ctaattcaat agcatatgtt acccaacggg aagcatatgc tatcgaatta
     3061 gggttagtaa aagggtccta aggaacagcg atatctccca ccccatgagc tgtcacggtt
     3121 ttatttacat ggggtcagga ttccacgagg gtagtgaacc attttagtca caagggcagt
     3181 ggctgaagat caaggagcgg gcagtgaact ctcctgaatc ttcgcctgct tcttcattct
     3241 ccttcgttta gctaatagaa taactgctga gttgtgaaca gtaaggtgta tgtgaggtgc
     3301 tcgaaaacaa ggtttcaggt gacgccccca gaataaaatt tggacggggg gttcagtggt
     3361 ggcattgtgc tatgacacca atataaccct cacaaacccc ttgggcaata aatactagtg
     3421 taggaatgaa acattctgaa tatctttaac aatagaaatc catggggtgg ggacaagccg
     3481 taaagactgg atgtccatct cacacgaatt tatggctatg ggcaacacat aatcctagtg
     3541 caatatgata ctggggttat taagatgtgt cccaggcagg gaccaagaca ggtgaaccat
     3601 gttgttacac tctatttgta acaaggggaa agagagtgga cgccgacagc agcggactcc
     3661 actggttgtc tctaacaccc ccgaaaatta aacggggctc cacgccaatg gggcccataa
     3721 acaaagacaa gtggccactc ttttttttga aattgtggag tgggggcacg cgtcagcccc
     3781 cacacgccgc cctgcggttt tggactgtaa aataagggtg taataacttg gctgattgta
     3841 accccgctaa ccactgcggt caaaccactt gcccacaaaa ccactaatgg caccccgggg
     3901 aatacctgca taagtaggtg ggcgggccaa gataggggcg cgattgctgc gatctggagg
     3961 acaaattaca cacacttgcg cctgagcgcc aagcacaggg ttgttggtcc tcatattcac
     4021 gaggtcgctg agagcacggt gggctaatgt tgccatgggt agcatatact acccaaatat
     4081 ctggatagca tatgctatcc taatctatat ctgggtagca taggctatcc taatctatat
     4141 ctgggtagca tatgctatcc taatctatat ctgggtagta tatgctatcc taatttatat
     4201 ctgggtagca taggctatcc taatctatat ctgggtagca tatgctatcc taatctatat
     4261 ctgggtagta tatgctatcc taatctgtat ccgggtagca tatgctatcc taatagagat
     4321 tagggtagta tatgctatcc taatttatat ctgggtagca tatactaccc aaatatctgg
     4381 atagcatatg ctatcctaat ctatatctgg gtagcatatg ctatcctaat ctatatctgg
     4441 gtagcatagg ctatcctaat ctatatctgg gtagcatatg ctatcctaat ctatatctgg
     4501 gtagtatatg ctatcctaat ttatatctgg gtagcatagg ctatcctaat ctatatctgg
     4561 gtagcatatg ctatcctaat ctatatctgg gtagtatatg ctatcctaat ctgtatccgg
     4621 gtagcatatg ctatcctcat gataagctgt caaacatgag aattttcttg aagacgaaag
     4681 ggcctcgtga tacgcctatt tttataggtt aatgtcatga taataatggt ttcttagacg
     4741 tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata
     4801 cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga
     4861 aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc ttattccctt ttttgcggca
     4921 ttttgccttc ctgtttttgc tcacccagaa acgctggtga aagtaaaaga tgctgaagat
     4981 cagttgggtg cacgagtggg ttacatcgaa ctggatctca acagcggtaa gatccttgag
     5041 agttttcgcc ccgaagaacg ttttccaatg atgagcactt ttaaagttct gctatgtggc
     5101 gcggtattat cccgtgttga cgccgggcaa gagcaactcg gtcgccgcat acactattct
     5161 cagaatgact tggttgagta ctcaccagtc acagaaaagc atcttacgga tggcatgaca
     5221 gtaagagaat tatgcagtgc tgccataacc atgagtgata acactgcggc caacttactt
     5281 ctgacaacga tcggaggacc gaaggagcta accgcttttt tgcacaacat gggggatcat
     5341 gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt
     5401 gacaccacga tgcctgcagc aatggcaaca acgttgcgca aactattaac tggcgaacta
     5461 cttactctag cttcccggca acaattaata gactggatgg aggcggataa agttgcagga
     5521 ccacttctgc gctcggccct tccggctggc tggtttattg ctgataaatc tggagccggt
     5581 gagcgtgggt ctcgcggtat cattgcagca ctggggccag atggtaagcc ctcccgtatc
     5641 gtagttatct acacgacggg gagtcaggca actatggatg aacgaaatag acagatcgct
     5701 gagataggtg cctcactgat taagcattgg taactgtcag accaagttta ctcatatata
     5761 ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt
     5821 gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc
     5881 gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg
     5941 caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact
     6001 ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt tcttctagtg
     6061 tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg
     6121 ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac
     6181 tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca
     6241 cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga
     6301 gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc
     6361 ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct
     6421 gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg
     6481 agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct
     6541 tttgctcaca tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc
     6601 tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc
     6661 gaggaagcgg aagagcgccc aatacgcaaa ccgcctctcc ccgcgcgttg gccgattcat
     6721 taatgcagct ggcacgacag gtttcccgac tggaaagcgg gcagtgagcg caacgcaatt
     6781 aatgtgagtt agctcactca ttaggcaccc caggctttac actttatgct tccggctcgt
     6841 atgttgtgtg gaattgtgag cggataacaa tttcacacag gaaacagcta tgaccatgat
     6901 tacgccaagc tctagctaga ggtcgaccaa ttctcatgtt tgacagctta tcatcgcaga
     6961 tccgggcaac gttgttgcca ttgctgcagg cgcagaactg gtaggtatgg aagatctata
     7021 cattgaatca atattggcaa ttagccatat tagtcattgg ttatatagca taaatcaata
     7081 ttggctattg gccattgcat acgttgtatc tatatcataa tatgtacatt tatattggct
     7141 catgtccaat atgaccgc
//
