LOCUS       sequence-242838-        9189 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 9189)
  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 9189)
  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_P92-Cd4-bio-His
FEATURES             Location/Qualifiers
     source          1..9189
                     /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             4353..4370
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    4422..4477
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(4476..4495)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(6713..6731)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6799..6903
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6904..6972,6973..7764)
                     /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(7122..7141)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7935..8523
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8424..8443
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     8677..8694
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    8811..8832
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(8847..8852,8853..8870,8871..8877)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8885..8901
                     /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     8890..8912
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     8909..8925
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     8909..8925
                     /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 atgtgatcct ggaaggcgag tttgagagca agctgggcta cgacggcgag cagatcgtgt
     1261 acagcaagaa gaacttcaag ggcagcatct acgccttcgt gcccgtgaag ggcaagtgcg
     1321 gcatcagcgt gcaggaaagc acagacggcg tcgagtggga gaacaacgcc gccatcgagg
     1381 tggaagatgg cgtgaacgag gccaccctgt tcagcgtgtt caccaccgac gacgccgtga
     1441 tcgtgatctt ccggtgcagc aacatctact atatcgccaa gcggggcgag ggcaacgcct
     1501 ggaagaacat caaacaggtg tccttcagcg acttccccgc cgacgccatc cctgccgtgt
     1561 actctggcag ctacctgctg atgaacaacg agtacgagaa gttcatcctc gtgtgcgcca
     1621 gagaggaccc caacggcgcc gaacacggca acgaggccga tagcgaagtg aaggccgtgc
     1681 ctttcagcga actgcagctg ctgggccggt gcaagttcag cttcgacgag ggcgagacat
     1741 ggcgcgaggg cgtgaccaga atccacagcg acgacctgag cagctatgac ggcggcgagg
     1801 atagcaccga gctgagccac gatgacgccc tgaagtccgt gaaagaagtg aacatggccg
     1861 aggaagagga agtcaagctg gaaggccagg gcaccgagga atccaaagag ggcgctcccc
     1921 tgcggaacgg catgcagatc agactgatcc agatgggcgg caacctggcc atccgggccg
     1981 tgaacattgc caaggacagc aacagcgcca agaccgtgat ccaggtctgc aacagcctgt
     2041 acaagaacga cctgtactgc gacaaagtga agctgcagat caaggacgag tacgtgctgc
     2101 acacctacga gatggtcaac ggctaccaca tggccgtgct gagagagaga aacggcgccc
     2161 acctggtgcc cagcttcgtg tacaatctgg ccgagacatt cgagcccatc ctgcactaca
     2221 aatacgagaa caatggggag ggcaagagcg acgtcgtggg cggcatccct ctggacaaga
     2281 ccacctatga gttcgtgctg gccaagggcg atatggtgtt cctgttctcc agcaccgtgg
     2341 ggaagaagaa caaagtcatc aagatcaacg cccctcggcg gaaggccgga tgcgaagtcc
     2401 tgcctgatgg cgagagcaac aagggctaca gccacacctt cagcatccgg aacatgcaga
     2461 accaccggat ctgcaagatg gccaccagcg agctgggaac caccgaggac ggcctgttca
     2521 aggtgttcta cgtgaccctg cccaagggcg tgtccctgaa cgacgactgc ttcagattca
     2581 gcttcagcaa agagctgaag tctgagtacc acaccaccgt ggtcaccaag agcaccaccg
     2641 gcaatgccgc tcacgaaggg gagatccagt ttagcttccc actgtactac agcaagtacc
     2701 tgtacaacta caagaaggcc tactgcaccc tgagcaacgg cctgatggtg gccgtggaat
     2761 tcgactacct gagaaacatc ctggacctga actacgccga cgagaacgac gtgatcaaga
     2821 ttaccagcaa cgacatcgtg gtgcatgcca agcggagata cgtccaggaa aagggcggcg
     2881 acatcttccc cagcggcacc tacatcaaca cctacctgcc cttcgagaaa gagtacgtca
     2941 tcagcaactt cgtccccaag tctgtgaatg aggtgtccac catcaccctg ccccacggct
     3001 ctgtgcggaa gctgcacttt ctgcagagcg gcgaggtgct gccctacgat ggcatcgacc
     3061 tgtccaacgt ggcccctaat tacaagaagc tgactaagta cgagctgaac cgcaagaccc
     3121 tggatgtgat catctccaac ttcaagtccg atgagaagat tatcggactc gtgtgccctc
     3181 tgagcagcga ggatgagggc ctgacctgct tcgacaaggt gtacctgcag aagggcggcc
     3241 tgcacaacat cgagttcgtg tttggcagca acgatatctt cgtgatccct cagcggcggc
     3301 tgctgaggcc tggcggcaaa atggccatgg aaagcctgct gtacctgaac aagcggaacg
     3361 tggacaagct gaagaaagag aagaatatca tccacttttt ctgcgagtgc aagggcgacg
     3421 aggacaccga gagcgtgcgc gtgaactact acatcagcgc ctactacgac agcgagtttg
     3481 tgaaggccca ggtcggaaag tccggcgagt ccagcttcat caacggccac atcatggtgc
     3541 agcgggaaga ggccggcaaa gaatccaccg tggataaggt ggacgacaag gtcgactgga
     3601 tcagcagcga caccaacctg aacatggaag agggcgtccg gatgatcggc cagaacatgc
     3661 tgtttagcat gctgagcggc ggcaacagca agggcaaccc tggcgcgccg tcgacctcca
     3721 tcacggccta taagagtgag ggggagtcag cggagttctc cttcccactc aaccttggag
     3781 aggaaagcct gcagggagag ttgagatgga aggcagagaa ggctccttct tcccagtcct
     3841 ggatcacctt ctccctaaag aaccaaaagg tgtctgtgca gaagtctact agcaacccca
     3901 agttccagct gtccgaaacg ctcccactca cccttcagat accccaggtc tcccttcagt
     3961 ttgctggttc tggcaacctg accctgactc tggacagagg gatactgtat caggaagtga
     4021 acctggtggt gatgaaagtg actcagcccg acagcaacac tttgacctgt gaggtgatgg
     4081 gacccacctc acccaagatg agactgatct tgaagcagga gaatcaggag gccagggtct
     4141 ccaggcagga gaaagtgatt caagtgcagg cccctgaagc aggggtgtgg caatgtctac
     4201 tgagtgaagg tgaagaggtc aagatggact ccaagatcca ggttttatcc aaagggttga
     4261 attccggcag cctgcaccac atcctggacg cccagaagat gctgtggaac cacagggaca
     4321 ggaacctgcc tcctctggcc cctctgggcc cccaccacca tcaccaccat tgagtctaga
     4381 cacgtgatta attaaggatc ccccgacctc gacctctggc taataaagga aatttatttt
     4441 cattgcaata gtgtgttgga attttttgtg tctctcactc ggaaggacat atgggagggc
     4501 aaatcatttg gtcgagatcc ctcggagatc tctagctaga ggatcgatcc ccgccccgga
     4561 cgaactaaac ctgactacga catctctgcc ccttcttcgc ggggcagtgc atgtaatccc
     4621 ttcagttggt tggtacaact tgccaactgg gccctgttcc acatgtgaca cgggggggga
     4681 ccaaacacaa aggggttctc tgactgtagt tgacatcctt ataaatggat gtgcacattt
     4741 gccaacactg agtggctttc atcctggagc agactttgca gtctgtggac tgcaacacaa
     4801 cattgccttt atgtgtaact cttggctgaa gctcttacac caatgctggg ggacatgtac
     4861 ctcccagggg cccaggaaga ctacgggagg ctacaccaac gtcaatcaga ggggcctgtg
     4921 tagctaccga taagcggacc ctcaagaggg cattagcaat agtgtttata aggccccctt
     4981 gttaacccta aacgggtagc atatgcttcc cgggtagtag tatatactat ccagactaac
     5041 cctaattcaa tagcatatgt tacccaacgg gaagcatatg ctatcgaatt agggttagta
     5101 aaagggtcct aaggaacagc gatatctccc accccatgag ctgtcacggt tttatttaca
     5161 tggggtcagg attccacgag ggtagtgaac cattttagtc acaagggcag tggctgaaga
     5221 tcaaggagcg ggcagtgaac tctcctgaat cttcgcctgc ttcttcattc tccttcgttt
     5281 agctaataga ataactgctg agttgtgaac agtaaggtgt atgtgaggtg ctcgaaaaca
     5341 aggtttcagg tgacgccccc agaataaaat ttggacgggg ggttcagtgg tggcattgtg
     5401 ctatgacacc aatataaccc tcacaaaccc cttgggcaat aaatactagt gtaggaatga
     5461 aacattctga atatctttaa caatagaaat ccatggggtg gggacaagcc gtaaagactg
     5521 gatgtccatc tcacacgaat ttatggctat gggcaacaca taatcctagt gcaatatgat
     5581 actggggtta ttaagatgtg tcccaggcag ggaccaagac aggtgaacca tgttgttaca
     5641 ctctatttgt aacaagggga aagagagtgg acgccgacag cagcggactc cactggttgt
     5701 ctctaacacc cccgaaaatt aaacggggct ccacgccaat ggggcccata aacaaagaca
     5761 agtggccact cttttttttg aaattgtgga gtgggggcac gcgtcagccc ccacacgccg
     5821 ccctgcggtt ttggactgta aaataagggt gtaataactt ggctgattgt aaccccgcta
     5881 accactgcgg tcaaaccact tgcccacaaa accactaatg gcaccccggg gaatacctgc
     5941 ataagtaggt gggcgggcca agataggggc gcgattgctg cgatctggag gacaaattac
     6001 acacacttgc gcctgagcgc caagcacagg gttgttggtc ctcatattca cgaggtcgct
     6061 gagagcacgg tgggctaatg ttgccatggg tagcatatac tacccaaata tctggatagc
     6121 atatgctatc ctaatctata tctgggtagc ataggctatc ctaatctata tctgggtagc
     6181 atatgctatc ctaatctata tctgggtagt atatgctatc ctaatttata tctgggtagc
     6241 ataggctatc ctaatctata tctgggtagc atatgctatc ctaatctata tctgggtagt
     6301 atatgctatc ctaatctgta tccgggtagc atatgctatc ctaatagaga ttagggtagt
     6361 atatgctatc ctaatttata tctgggtagc atatactacc caaatatctg gatagcatat
     6421 gctatcctaa tctatatctg ggtagcatat gctatcctaa tctatatctg ggtagcatag
     6481 gctatcctaa tctatatctg ggtagcatat gctatcctaa tctatatctg ggtagtatat
     6541 gctatcctaa tttatatctg ggtagcatag gctatcctaa tctatatctg ggtagcatat
     6601 gctatcctaa tctatatctg ggtagtatat gctatcctaa tctgtatccg ggtagcatat
     6661 gctatcctca tgataagctg tcaaacatga gaattttctt gaagacgaaa gggcctcgtg
     6721 atacgcctat ttttataggt taatgtcatg ataataatgg tttcttagac gtcaggtggc
     6781 acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat
     6841 atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag
     6901 agtatgagta ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt
     6961 cctgtttttg ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt
     7021 gcacgagtgg gttacatcga actggatctc aacagcggta agatccttga gagttttcgc
     7081 cccgaagaac gttttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta
     7141 tcccgtgttg acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac
     7201 ttggttgagt actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa
     7261 ttatgcagtg ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg
     7321 atcggaggac cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc
     7381 cttgatcgtt gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg
     7441 atgcctgcag caatggcaac aacgttgcgc aaactattaa ctggcgaact acttactcta
     7501 gcttcccggc aacaattaat agactggatg gaggcggata aagttgcagg accacttctg
     7561 cgctcggccc ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggg
     7621 tctcgcggta tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc
     7681 tacacgacgg ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt
     7741 gcctcactga ttaagcattg gtaactgtca gaccaagttt actcatatat actttagatt
     7801 gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc
     7861 atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag
     7921 atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa
     7981 aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg
     8041 aaggtaactg gcttcagcag agcgcagata ccaaatactg ttcttctagt gtagccgtag
     8101 ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg
     8161 ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga
     8221 tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc
     8281 ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc
     8341 acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga
     8401 gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt
     8461 cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg
     8521 aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac
     8581 atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc ctttgagtga
     8641 gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg
     8701 gaagagcgcc caatacgcaa accgcctctc cccgcgcgtt ggccgattca ttaatgcagc
     8761 tggcacgaca ggtttcccga ctggaaagcg ggcagtgagc gcaacgcaat taatgtgagt
     8821 tagctcactc attaggcacc ccaggcttta cactttatgc ttccggctcg tatgttgtgt
     8881 ggaattgtga gcggataaca atttcacaca ggaaacagct atgaccatga ttacgccaag
     8941 ctctagctag aggtcgacca attctcatgt ttgacagctt atcatcgcag atccgggcaa
     9001 cgttgttgcc attgctgcag gcgcagaact ggtaggtatg gaagatctat acattgaatc
     9061 aatattggca attagccata ttagtcattg gttatatagc ataaatcaat attggctatt
     9121 ggccattgca tacgttgtat ctatatcata atatgtacat ttatattggc tcatgtccaa
     9181 tatgaccgc
//
