LOCUS       sequence-297679-        4075 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Cas9 and gRNA expression plasmid for P. falciparum with yDHODH 
            selectable marker..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 4075)
  AUTHORS   Nessel T, Beck JM, Rayatpisheh S, Jami-Alahmadi Y, Wohlschlegel JA, 
            Goldberg DE, Beck JR
  TITLE     EXP1 is required for organisation of EXP2 in the intraerythrocytic 
            malaria parasite vacuole.
  JOURNAL   Cell Microbiol. 2020 Jan 28:e13168. doi: 10.1111/cmi.13168.
  PUBMED    31990132
REFERENCE   2  (bases 1 to 4075)
  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: "Cell 
            Microbiol. 2020 Jan 28:e13168. doi: 10.1111/cmi.13168."
COMMENT     Sequence Label: pUF-Cas9-pre-sgRNA
FEATURES             Location/Qualifiers
     source          1..4075
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     primer_bind     complement(1700..1719)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     1819..1841
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(1879..1897)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1965..2069
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2070..2138,2139..2930)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(2288..2307)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3101..3689
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3590..3609
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3843..3860
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    3977..3998
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(4013..4018,4019..4036,4037..4043)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4051..4067
                     /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)."
ORIGIN
        1 attttattta ttatcattca agatttatat tttataataa taaataccta atagaaatat
       61 atcaggatcc ttcgaaatga cagccagttt aactaccaag ttcttgaaca atacctatga
      121 aaacccattt atgaatgcat ccggtgttca ttgcatgact acacaagaat tagatgaatt
      181 agcaaactct aaagctggcg cattcattac aaagagtgct acaaccttag aaagagaagg
      241 taaccctgaa ccacgttaca tttctgtccc tctaggcagt atcaactcaa tgggtttacc
      301 aaacgaaggt atcgactact atttgtccta tgtattaaac cgtcaaaaga attatcctga
      361 tgcacctgct attttcttct cagttgctgg tatgagcatt gatgaaaatt taaatttgtt
      421 gaggaaaatc caagatagcg aatttaacgg tattaccgag ttaaacttgt cttgtcctaa
      481 tgtgcctggg aaaccacaag ttgcttatga ctttgacttg acaaaggaaa ccttggaaaa
      541 ggtttttgcc tttttcaaaa aacctcttgg tgtcaagttg cctccttatt ttgattttgc
      601 ccattttgat atcatggcaa aaatattgaa cgagttccca ttagcttatg tcaactctat
      661 caatagtata ggaaatggtc ttttcattga tgtggagaag gagagtgtag tagtgaagcc
      721 aaagaatggt ttcgggggta ttggaggtga atatgttaag ccaaccgcgc tcgccaatgt
      781 tcgtgcattt tacactcgtt tgagacctga aatcaaagtt atcggtacag gtggaattaa
      841 gtccggtaag gatgcatttg aacatcttct atgtggtgcc tctatgctac agattggtac
      901 agaattacaa aaagagggcg tcaagatttt tgaacgtatc gaaaaagaat taaaagacat
      961 aatggaagct aagggttata catccataga tcagttccgt gggaagttga acagcattta
     1021 aaagcttatt taataataga ttaaaaatat tataaaaata aaaacataaa cacagaaatt
     1081 acaaaaaaaa tacatatgaa tttttttttg taatcttcct tataaatata gaataatgaa
     1141 tcatataaaa catatcatta ttcatttatt tacatttaaa attattgttt cagtatcttt
     1201 aatttattat gtatatataa aaataactta caattttatt aataagcaat atatgtttat
     1261 taattcatgt tttgtaattt atgggatagc gatttttttt actgtctgta tttttctttt
     1321 ttaattatgt tttaattgta ttttattttt attattgttc tttttatagt attattttaa
     1381 aacaaaatgt attttctaag aacttataat aataataaat ataaatttta ataaaaatta
     1441 tatttatctt ttacaatatg aacataaagt acaacattaa tatatagctt ttaatatttt
     1501 tattcctaat catgtaaatc ttaaattttt ctttttaaac atatgttaaa tatttatttc
     1561 tcattatata taagaacata tttattaaat ctagaattca tatcgataac tccatggaac
     1621 tcctaggctg gcgcctgatg cggtattttc tccttacgca tctgtgcggt atttcacacc
     1681 gcatatggtg cactctcagt acaatctgct ctgatgccgc atagttaagc cagccccgac
     1741 acccgccaac acccgctgac gcgccctgac gggcttgtct gctcccggca tccgcttaca
     1801 gacaagctgt gaccgtctcc gggagctgca tgtgtcagag gttttcaccg tcatcaccga
     1861 aacgcgcgag acgaaagggc ctcgtgatac gcctattttt ataggttaat gtcatgataa
     1921 taatggtttc ttagacgtca ggtggcactt ttcggggaaa tgtgcgcgga acccctattt
     1981 gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa
     2041 tgcttcaata atattgaaaa aggaagagta tgagtattca acatttccgt gtcgccctta
     2101 ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg ctggtgaaag
     2161 taaaagatgc tgaagatcag ttgggtgcac gagtgggtta catcgaactg gatctcaaca
     2221 gcggtaagat ccttgagagt tttcgccccg aagaacgttt tccaatgatg agcactttta
     2281 aagttctgct atgtggcgcg gtattatccc gtattgacgc cgggcaagag caactcggtc
     2341 gccgcataca ctattctcag aatgacttgg ttgagtactc accagtcaca gaaaagcatc
     2401 ttacggatgg catgacagta agagaattat gcagtgctgc cataaccatg agtgataaca
     2461 ctgcggccaa cttacttctg acaacgatcg gaggaccgaa ggagctaacc gcttttttgc
     2521 acaacatggg ggatcatgta actcgccttg atcgttggga accggagctg aatgaagcca
     2581 taccaaacga cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac
     2641 tattaactgg cgaactactt actctagctt cccggcaaca attaatagac tggatggagg
     2701 cggataaagt tgcaggacca cttctgcgct cggcccttcc ggctggctgg tttattgctg
     2761 ataaatctgg agccggtgag cgtgggtctc gcggtatcat tgcagcactg gggccagatg
     2821 gtaagccctc ccgtatcgta gttatctaca cgacggggag tcaggcaact atggatgaac
     2881 gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggtaa ctgtcagacc
     2941 aagtttactc atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct
     3001 aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc
     3061 actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc
     3121 gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg
     3181 atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa
     3241 atactgttct tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc
     3301 ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt
     3361 gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa
     3421 cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc
     3481 tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc
     3541 cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct
     3601 ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat
     3661 gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc
     3721 tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg
     3781 ataaccgtat taccgccttt gagtgagctg ataccgctcg ccgcagccga acgaccgagc
     3841 gcagcgagtc agtgagcgag gaagcggaag agcgcccaat acgcaaaccg cctctccccg
     3901 cgcgttggcc gattcattaa tgcagctggc acgacaggtt tcccgactgg aaagcgggca
     3961 gtgagcgcaa cgcaattaat gtgagttagc tcactcatta ggcaccccag gctttacact
     4021 ttatgcttcc ggctcgtatg ttgtgtggaa ttgtgagcgg ataacaattt cacac
//
