LOCUS       sequence-357204-        7639 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  P. berghei D3 (502-617) construct with N516T, S533N and N539Q 
            mutations to abolish N-glycosylation was used for crystallization..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7639)
  AUTHORS   Feng J, Dong X, DeCosta A, Su Y, Angrisano F, Sala KA, Blagborough 
            AM, Lu C, Springer TA
  TITLE     Structural basis of malaria transmission blockade by a monoclonal 
            antibody to gamete fusogen HAP2.
  JOURNAL   Elife. 2021 Dec 23;10. pii: 74707. doi: 10.7554/eLife.74707.
  PUBMED    34939934
REFERENCE   2  (bases 1 to 7639)
  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: "Elife."; 
            date: "2021-12-23"; volume: "10. pii: 74707. doi"; pages: " 
            10.7554/eLife.74707"
COMMENT     Sequence Label: Pb mD3S4-dN
FEATURES             Location/Qualifiers
     source          1..7639
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        3..306
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        308..583
                     /label=chicken beta-actin promoter
     intron          584..1600
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1524..1546
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1608..1627
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     CDS             2113..2133
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=7xHis
                     /translation="HHHHHHH"
     polyA_signal    3710..3765
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(3764..3783)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     rep_origin      complement(4475..5063)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4555..4574)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(5149..5958)
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP
                     DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA
                     FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
                     FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
                     RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     5872..5891
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     promoter        complement(5977..6024)
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             complement(6450..7049)
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(6533..6553)
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     7030..7049
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     promoter        complement(7059..7558)
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(7107..7126)
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     primer_bind     7500..7523
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
ORIGIN
        1 cgcgttacat aacttacggt aaatggcccg cctggctgac cgcccaacga cccccgccca
       61 ttgacgtcaa taatgacgta tgttcccata gtaacgccaa tagggacttt ccattgacgt
      121 caatgggtgg agtatttacg gtaaactgcc cacttggcag tacatcaagt gtatcatatg
      181 ccaagtacgc cccctattga cgtcaatgac ggtaaatggc ccgcctggca ttatgcccag
      241 tacatgacct tatgggactt tcctacttgg cagtacatct acgtattagt catcgctatt
      301 accatggtcg aggtgagccc cacgttctgc ttcactctcc ccatctcccc cccctcccca
      361 cccccaattt tgtatttatt tattttttaa ttattttgtg cagcgatggg ggcggggggg
      421 gggggggcgc gcgccaggcg gggcggggsg gggsgrgggg sggggsgggg sgaggcggrg
      481 aggkgcggcg gcagccaatc agagcggcgc gctccgaaag tttcctttta tggcgaggcg
      541 gcggcggcgg cggccctata aaaagcgaag cgcgcggcgg gcgggagtcg ctgcgcgctg
      601 ccttcgcccc gtgccccgct ccgccgccgc ctcgcgccgc ccgccccggc tctgactgac
      661 cgcgttactc ccacaggtga gcgggcggga cggcccttct cctccgggct gtaattagcg
      721 cttggtttaa tgacggcttg tttcttttct gtggctgcgt gaaagccttg aggggctccg
      781 ggagggccct ttgtgcgggg ggagcggctc ggggggtgcg tgcgtgtgtg tgtgcgtggg
      841 gagcgccgcg tgcggctccg cgctgcccgg cggctgtgag cgctgcgggc gcggcgcggg
      901 gctttgtgcg ctccgcagtg tgcgcgaggg gagcgcggcc gggggcggtg ccccgcggtg
      961 cggggggggc tgcgagggga acaaaggctg cgtgcggggt gtgtgcgtgg gggggtgagc
     1021 agggggtgtg ggcgcgtcgg tcgggctgca accccccctg cacccccctc cccgagttgc
     1081 tgagcacggc ccggcttcgg gtgcggggct ccgtacgggg cgtggcgcgg ggctcgccgt
     1141 gccgggcggg gggtggcggc aggtgggggt gccgggcggg gcggggccgc ctcgggccgg
     1201 ggagggctcg ggggaggggc gcggcggccc ccggagcgcc ggcggctgtc gaggcgcggc
     1261 gagccgcagc cattgccttt tatggtaatc gtgcgagagg gcgcagggac ttcctttgtc
     1321 ccaaatctgt gcggagccga aatctgggag gcgccgccgc accccctcta gcgggcgcgg
     1381 ggcgaagcgg tgcggcgccg gcaggaagga aatgggcggg gagggccttc gtgcgtcgcc
     1441 gcgccgccgt ccccttctcc ctctccagcc tcggggctgt ccgcgggggg acggctgcct
     1501 tcggggggga cggggcaggg cggggttcgg cttctggcgt gtgaccggcg gctctagagc
     1561 ctctgctaac catgttcatg ccttcttctt tttcctacag ctcctgggca acgtgctggt
     1621 tattgtgctg tctcatcatt ttggcaaaga attcggcttg atcgaagccg tctcagggga
     1681 gagaccacac ccaagctgtc tagagccgcc accatgcctt tgctcctcct gttgcccttg
     1741 ctctgggctg gcgccctcgc ggccgccacc atcacgcacg tcacgattcc gaatgactgt
     1801 gccagcacca actccaattc caatgagtgc gttctgatca ttcatgtctg gaacaataac
     1861 aagttcgtgg gcagtcagtt cagctgctct atcgcgtgca ctaacaagga gactgatcaa
     1921 cttgcatcgc acatcaatcc catagcccca gtacgcgcct tcatcggtcc caacaaaaac
     1981 tacgcatttt acttcatcat taagttcctc atcaataagg agatcacaac gctgtgcaag
     2041 gccatcgtga aggactccaa cggaaaggag tgcagtatag aggaatttga gttgcaatct
     2101 aaggaatccg tccatcacca ccatcaccat cattaaagta cttgaggtct ctaaaagcgt
     2161 cttcctgttc tcatcacatc atatcaaggt tatataccat caatattgcc acagatgtta
     2221 cttagccttt taatatttct ctaatttagt gtatatgcaa tgatagttct ctgatttctg
     2281 agattgagtt tctcatgtgt aatgattatt tagagtttct ctttcatctg ttcaaatttt
     2341 tgtctagttt tattttttac tgatttgtaa gacttctttt tataatctgc atattacaat
     2401 tctctttact ggggtgttgc aaatattttc tgtcattcta tggcctgact tttcttaatg
     2461 gttttttaat tttaaaaata agtcttaata ttcatgcaat ctaattaaca atcttttctt
     2521 tgtggttagg actttgagtc ataagaaatt tttctctaca ctgaagtcat gatggcatgc
     2581 ttctatatta ttttctaaaa gatttaaagt tttgccttct ccatttagac ttataattca
     2641 ctggaatttt tttgtgtgta tggtatgaca tatgggttcc cttttatttt ttacatataa
     2701 atatatttcc ctgtttttct aaaaaagaaa aagatcatca ttttcccatt gtaaaatgcc
     2761 atattttttt cataggtcac ttacatatat caatgggtct gtttctgagc tctactctat
     2821 tttatcagcc tcactgtcta tccccacaca tctcatgctt tgctctaaat cttgatattt
     2881 agtggaacat tctttcccat tttgttctac aagaatattt ttgttattgt ctttgggctt
     2941 tctatataca ttttgaaatg aggttgacaa gttaataaca ggcctattga ttggaaagtt
     3001 tgtcaacgaa ttgtgggtct tttggggttt gctgcccctt ttacgcaatg tggatatcct
     3061 gctttaatgc ctttatatgc atgtatacaa gcaaaacagg cttttacttt ctcgccaact
     3121 tacaaggcct ttctcagtaa acagtatatg accctttacc ccgttgctcg gcaacggcct
     3181 ggtctgtgcc aagtgtttgc tgacgcaacc cccactggtt ggggcttggc cataggccat
     3241 cagcgcatgc gtggaacctt tgtgtctcct ctgccgatcc atactgcgga actcctagcc
     3301 gcttgttttg ctcgcagcag gtctggagca aacctcatcg ggaccgacaa ttctgtcgta
     3361 ctctcccgca agtatacatc gtttccatgg ctgctaggct gtgctgccaa ctggatcctg
     3421 cgcgggacgt cctttgttta cgtcccgtcg gcgctgaatc ccgcggacga cccctcccgg
     3481 ggccgcttgg ggctctaccg cccgcttctc cgtctgccgt accgtccgac cacggggcgc
     3541 acctctcttt acgcggactc cccgtctgtg ccttctcatc tgccggaccg tgtgcacttc
     3601 gcttcacctc tgcacgtcgc atggaggcca ccgtgaacgc ccaccggaac ctgcccaagg
     3661 tcttgcataa gaggactctt ggactttcag caatgtcatc tcagtggcta ataaaggaaa
     3721 tttattttca ttgcaatagt gtgttggaat tttttgtgtc tctcactcgg aaggacatat
     3781 gggagggcaa atcatttaaa acatcagaat gagtatttgg tttagagttt ggcaacatat
     3841 gcccatatgc tggctgccat gaacaaaggt tggctataaa gaggtcatca gtatatgaaa
     3901 cagccccctg ctgtccattc cttattccat agaaaagcct tgacttgagg ttagattttt
     3961 tttatatttt gttttgtgtt atttttttct ttaacatccc taaaattttc cttacatgtt
     4021 ttactagcca gatttttcct cctctcctga ctactcccag tcatagctgt ccctcttctc
     4081 ttatggagat ctttgcattt ttagacattt agaagcctat atcttgttac agaattggaa
     4141 ttacacaaaa attctaccat attttgaaag cttaggttgt tctgaaaaaa acaatatatt
     4201 gttttcctgg gtaaactaaa agtcccctcg aggaaaggcc cctaaagtga aacagtgcaa
     4261 aacgttcaaa aactgtctgg caatacaagt tccactttga ccaaaacggc tggcagtaaa
     4321 agggttaaga agactgtcag ccttgagcgg tatcagctca ctcaaaggcg gtaatacggt
     4381 tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg
     4441 ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg
     4501 agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat
     4561 accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta
     4621 ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct
     4681 gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc
     4741 ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa
     4801 gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg
     4861 taggcggtgc tacagagttc ttgaagtggt gggctaacta cggctacact agaagaacag
     4921 tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt
     4981 gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta
     5041 cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc
     5101 agtggaacga cgcgcgcgta actcacgtta agggattttg gtcatgagtt agaaaaactc
     5161 atcgagcatc aaatgaaact gcaatttatt catatcagga ttatcaatac catatttttg
     5221 aaaaagccgt ttctgtaatg aaggagaaaa ctcaccgagg cagttccata ggatggcaag
     5281 atcctggtat cggtctgcga ttccgactcg tccaacatca atacaaccta ttaatttccc
     5341 ctcgtcaaaa ataaggttat caagtgagaa atcaccatga gtgacgactg aatccggtga
     5401 gaatggcaaa agtttatgca tttctttcca gacttgttca acaggccagc cattacgctc
     5461 gtcatcaaaa tcactcgcat caaccaaacc gttattcatt cgtgattgcg cctgagcgag
     5521 gcgaaatacg cgatcgctgt taaaaggaca attacaaaca ggaatcgagt gcaaccggcg
     5581 caggaacact gccagcgcat caacaatatt ttcacctgaa tcaggatatt cttctaatac
     5641 ctggaacgct gtttttccgg ggatcgcagt ggtgagtaac catgcatcat caggagtacg
     5701 gataaaatgc ttgatggtcg gaagtggcat aaattccgtc agccagttta gtctgaccat
     5761 ctcatctgta acatcattgg caacgctacc tttgccatgt ttcagaaaca actctggcgc
     5821 atcgggcttc ccatacaagc gatagattgt cgcacctgat tgcccgacat tatcgcgagc
     5881 ccatttatac ccatataaat cagcatccat gttggaattt aatcgcggcc tcgacgtttc
     5941 ccgttggata tggctcattt tttacttcct caccttgtcg tattatacta tgccgatata
     6001 ctatgccgat gattaattgt cgacactgcg ggggctctgt gtggtaagca ggtcttaacc
     6061 tttttactgc caatgacgca tgggatacgt cgtggcagta aaagggctta aatgccaacg
     6121 acgcgtccca tacgttgttg gcattttaat tcttctctct gcagcggcag catgtgccgc
     6181 cgctgcagag agtttctagc gatgacagcc cctctgggca acgagccggg ggggctgtct
     6241 ttctttatgt tttaaatgca ctgacctccc acattccctt tttagtaaaa tattcagaaa
     6301 taatttaaat acatcattgc aatgaaaata aatgtttttt attaggcaga atccagatgc
     6361 tcaaggccct tcataatatc ccccagttta gtagttggac ttagggaaca aaggaacctt
     6421 taatagaaat tggacagcaa gaaagcgagt caggcaccgg gcttgcgggt catgcaccag
     6481 gtgcgcggtc cttcgggcac ctcgacgtcg gcggtgacgg tgaagccgag ccgctcgtag
     6541 aaggggaggt tgcggggcgc ggatgtctcc aggaaggcgg gcaccccggc gcgctcggcc
     6601 gcctccactc cggggagcac gacggcgctg cccagaccct tgccctggtg gtcgggcgac
     6661 acgccgacgg tggccaggaa ccacgcgggc tccttgggcc ggtgcggcgc caggaggcct
     6721 tccatctgtt gctgcgcggc cagccgggaa ccgctcaact cggccatgcg cgggccgatc
     6781 tcggcgaaca ccgcccccgc ttcgacgctc tccggcgtgg tccagaccgc caccgcggcg
     6841 ccgtcgtccg cgacccacac cttgccgatg tcgagcccga cgcgcgtgag gaagagttct
     6901 tgcagctcgg tgacccgctc gatgtggcgg tccggatcga cggtgtggcg cgtggcgggg
     6961 tagtcggcga acgcggcggc gagggtgcgt acggccctgg ggacgtcgtc gcgggtggcg
     7021 aggcgcaccg tgggcttgta ctcggtcatg gtggcggacg aaaggcccgg agatgaggaa
     7081 gaggagaaca gcgcggcaga cgtgcgcttt tgaagcgtgc agaatgccgg gcctccggag
     7141 gaccttcggg cgcccgcccc gcccctgagc ccgcccctga gcccgccccc ggacccaccc
     7201 cttcccagcc tctgagccca gaaagcgaag gagcaaagct gctattggcc gctgccccaa
     7261 aggcctaccc gcttccattg ctcagcggtg ctgtccatct gcacgagact agtgagtcgt
     7321 gctacttcca tttgtcacgt cctgcacgac gcgagctgcg gggcgggggg gaacttcctg
     7381 actaggggag gagtagaagg tggcgcgaag gggccaccaa agaacggagc cggttggcgc
     7441 ctaccggtgg atgtggaatg tgtgcgaggc cagaggccac ttgtgtagcg ccaagtgccc
     7501 agcggggctg ctaaagcgca tgctccagac tgccttggga aaagcgcctc ccctacccgg
     7561 tagagaaact tgatctgtcg ccgcaattca aacttcgtga ggctccggtg cccgtcagtg
     7621 acctgctata ctctggaga
//
