LOCUS       sequence-242731-        7707 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Expression of recombinant P. ovale protein in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7707)
  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 7707)
  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: Po_CyRPA-Cd4-bio-His
FEATURES             Location/Qualifiers
     source          1..7707
                     /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             2871..2888
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    2940..2995
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(2994..3013)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(5231..5249)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5317..5421
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5422..5490,5491..6282)
                     /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(5640..5659)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6453..7041
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6942..6961
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7195..7212
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    7329..7350
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7365..7370,7371..7388,7389..7395)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7403..7419
                     /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     7408..7430
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7427..7443
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7427..7443
                     /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 gcggccgcca
     1201 gctctacaaa gaatggcgcc gtgatcctga tcaacgacga catcagcacc gtgaccgacg
     1261 agaagtactg tatgcaggac agctacttcc tgttccacga cgagctgtac aagatctgca
     1321 tccagcacct catcaagaac gacatcgaga tcagcatcta cgtgcagaaa gaagtgaaga
     1381 acagctggga gacacaggaa gaactgatca gcgacgtgaa caccaagaag cccttcagcc
     1441 tgcacaccta catcgacgac aatgagatcg tgatcatcat gtgcaaatgc gtgaacgacc
     1501 tgagcatcga ccggaacgag tgctacagat gggacagcac caacggcaat ttcttcaaga
     1561 aagaggacat caccatcccc gacagcgcct tcaacaagcg gcggatgacc agctacaccc
     1621 tgtaccctct gaccatctcc accaagaaaa tcctgctgct gtgcggcatc aacatcgacc
     1681 cctacgtgcc ccagaagaaa gaagagttca tctactgcat cggcagcgag gaccggggca
     1741 agacctggat gcggaaggtg cagatcatct acgagggctt ccagcccaat ggcatctact
     1801 accacctgac ccctcacatc ttcaacgatg agctgggctt ccacttctac agcctcgtgg
     1861 gcgccaacaa tgctggcgtg ggcggcgagt acatcgtgtg caagcctgag aagggcatgc
     1921 tgtaccggat gaagtgcgag aacgccaacc tgggcaaaga gggcaagagc ctgcaggacc
     1981 tgaccaagat gaacggctac tatatcgcct cctacgtgaa caaagaaaag tccaatgagt
     2041 gctacctgta ctacaccagc gagaatgcca tcatcctgaa ccccaagaaa cagggctacc
     2101 tgatgaatgg ctgctacagc ggcttcttcg tgaagatcaa cgaggccagc accctgttta
     2161 tctacgcccc tgactacggc acctacaaca tccacaccat caagtacatg cgctacgatg
     2221 gcgcgccgtc gacctccatc acggcctata agagtgaggg ggagtcagcg gagttctcct
     2281 tcccactcaa ccttggagag gaaagcctgc agggagagtt gagatggaag gcagagaagg
     2341 ctccttcttc ccagtcctgg atcaccttct ccctaaagaa ccaaaaggtg tctgtgcaga
     2401 agtctactag caaccccaag ttccagctgt ccgaaacgct cccactcacc cttcagatac
     2461 cccaggtctc ccttcagttt gctggttctg gcaacctgac cctgactctg gacagaggga
     2521 tactgtatca ggaagtgaac ctggtggtga tgaaagtgac tcagcccgac agcaacactt
     2581 tgacctgtga ggtgatggga cccacctcac ccaagatgag actgatcttg aagcaggaga
     2641 atcaggaggc cagggtctcc aggcaggaga aagtgattca agtgcaggcc cctgaagcag
     2701 gggtgtggca atgtctactg agtgaaggtg aagaggtcaa gatggactcc aagatccagg
     2761 ttttatccaa agggttgaat tccggcagcc tgcaccacat cctggacgcc cagaagatgc
     2821 tgtggaacca cagggacagg aacctgcctc ctctggcccc tctgggcccc caccaccatc
     2881 accaccattg agtctagaca cgtgattaat taaggatccc ccgacctcga cctctggcta
     2941 ataaaggaaa tttattttca ttgcaatagt gtgttggaat tttttgtgtc tctcactcgg
     3001 aaggacatat gggagggcaa atcatttggt cgagatccct cggagatctc tagctagagg
     3061 atcgatcccc gccccggacg aactaaacct gactacgaca tctctgcccc ttcttcgcgg
     3121 ggcagtgcat gtaatccctt cagttggttg gtacaacttg ccaactgggc cctgttccac
     3181 atgtgacacg gggggggacc aaacacaaag gggttctctg actgtagttg acatccttat
     3241 aaatggatgt gcacatttgc caacactgag tggctttcat cctggagcag actttgcagt
     3301 ctgtggactg caacacaaca ttgcctttat gtgtaactct tggctgaagc tcttacacca
     3361 atgctggggg acatgtacct cccaggggcc caggaagact acgggaggct acaccaacgt
     3421 caatcagagg ggcctgtgta gctaccgata agcggaccct caagagggca ttagcaatag
     3481 tgtttataag gcccccttgt taaccctaaa cgggtagcat atgcttcccg ggtagtagta
     3541 tatactatcc agactaaccc taattcaata gcatatgtta cccaacggga agcatatgct
     3601 atcgaattag ggttagtaaa agggtcctaa ggaacagcga tatctcccac cccatgagct
     3661 gtcacggttt tatttacatg gggtcaggat tccacgaggg tagtgaacca ttttagtcac
     3721 aagggcagtg gctgaagatc aaggagcggg cagtgaactc tcctgaatct tcgcctgctt
     3781 cttcattctc cttcgtttag ctaatagaat aactgctgag ttgtgaacag taaggtgtat
     3841 gtgaggtgct cgaaaacaag gtttcaggtg acgcccccag aataaaattt ggacgggggg
     3901 ttcagtggtg gcattgtgct atgacaccaa tataaccctc acaaacccct tgggcaataa
     3961 atactagtgt aggaatgaaa cattctgaat atctttaaca atagaaatcc atggggtggg
     4021 gacaagccgt aaagactgga tgtccatctc acacgaattt atggctatgg gcaacacata
     4081 atcctagtgc aatatgatac tggggttatt aagatgtgtc ccaggcaggg accaagacag
     4141 gtgaaccatg ttgttacact ctatttgtaa caaggggaaa gagagtggac gccgacagca
     4201 gcggactcca ctggttgtct ctaacacccc cgaaaattaa acggggctcc acgccaatgg
     4261 ggcccataaa caaagacaag tggccactct tttttttgaa attgtggagt gggggcacgc
     4321 gtcagccccc acacgccgcc ctgcggtttt ggactgtaaa ataagggtgt aataacttgg
     4381 ctgattgtaa ccccgctaac cactgcggtc aaaccacttg cccacaaaac cactaatggc
     4441 accccgggga atacctgcat aagtaggtgg gcgggccaag ataggggcgc gattgctgcg
     4501 atctggagga caaattacac acacttgcgc ctgagcgcca agcacagggt tgttggtcct
     4561 catattcacg aggtcgctga gagcacggtg ggctaatgtt gccatgggta gcatatacta
     4621 cccaaatatc tggatagcat atgctatcct aatctatatc tgggtagcat aggctatcct
     4681 aatctatatc tgggtagcat atgctatcct aatctatatc tgggtagtat atgctatcct
     4741 aatttatatc tgggtagcat aggctatcct aatctatatc tgggtagcat atgctatcct
     4801 aatctatatc tgggtagtat atgctatcct aatctgtatc cgggtagcat atgctatcct
     4861 aatagagatt agggtagtat atgctatcct aatttatatc tgggtagcat atactaccca
     4921 aatatctgga tagcatatgc tatcctaatc tatatctggg tagcatatgc tatcctaatc
     4981 tatatctggg tagcataggc tatcctaatc tatatctggg tagcatatgc tatcctaatc
     5041 tatatctggg tagtatatgc tatcctaatt tatatctggg tagcataggc tatcctaatc
     5101 tatatctggg tagcatatgc tatcctaatc tatatctggg tagtatatgc tatcctaatc
     5161 tgtatccggg tagcatatgc tatcctcatg ataagctgtc aaacatgaga attttcttga
     5221 agacgaaagg gcctcgtgat acgcctattt ttataggtta atgtcatgat aataatggtt
     5281 tcttagacgt caggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt
     5341 ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa
     5401 taatattgaa aaaggaagag tatgagtatt caacatttcc gtgtcgccct tattcccttt
     5461 tttgcggcat tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat
     5521 gctgaagatc agttgggtgc acgagtgggt tacatcgaac tggatctcaa cagcggtaag
     5581 atccttgaga gttttcgccc cgaagaacgt tttccaatga tgagcacttt taaagttctg
     5641 ctatgtggcg cggtattatc ccgtgttgac gccgggcaag agcaactcgg tcgccgcata
     5701 cactattctc agaatgactt ggttgagtac tcaccagtca cagaaaagca tcttacggat
     5761 ggcatgacag taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc
     5821 aacttacttc tgacaacgat cggaggaccg aaggagctaa ccgctttttt gcacaacatg
     5881 ggggatcatg taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac
     5941 gacgagcgtg acaccacgat gcctgcagca atggcaacaa cgttgcgcaa actattaact
     6001 ggcgaactac ttactctagc ttcccggcaa caattaatag actggatgga ggcggataaa
     6061 gttgcaggac cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct
     6121 ggagccggtg agcgtgggtc tcgcggtatc attgcagcac tggggccaga tggtaagccc
     6181 tcccgtatcg tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga
     6241 cagatcgctg agataggtgc ctcactgatt aagcattggt aactgtcaga ccaagtttac
     6301 tcatatatac tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag
     6361 atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg
     6421 tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc
     6481 tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag
     6541 ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtt
     6601 cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac
     6661 ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc
     6721 gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt
     6781 tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt
     6841 gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc
     6901 ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt
     6961 tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca
     7021 ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt
     7081 tgctggcctt ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt
     7141 attaccgcct ttgagtgagc tgataccgct cgccgcagcc gaacgaccga gcgcagcgag
     7201 tcagtgagcg aggaagcgga agagcgccca atacgcaaac cgcctctccc cgcgcgttgg
     7261 ccgattcatt aatgcagctg gcacgacagg tttcccgact ggaaagcggg cagtgagcgc
     7321 aacgcaatta atgtgagtta gctcactcat taggcacccc aggctttaca ctttatgctt
     7381 ccggctcgta tgttgtgtgg aattgtgagc ggataacaat ttcacacagg aaacagctat
     7441 gaccatgatt acgccaagct ctagctagag gtcgaccaat tctcatgttt gacagcttat
     7501 catcgcagat ccgggcaacg ttgttgccat tgctgcaggc gcagaactgg taggtatgga
     7561 agatctatac attgaatcaa tattggcaat tagccatatt agtcattggt tatatagcat
     7621 aaatcaatat tggctattgg ccattgcata cgttgtatct atatcataat atgtacattt
     7681 atattggctc atgtccaata tgaccgc
//
