LOCUS       sequence-172843-        7486 bp    DNA     linear   UNA 14-APR-2025
DEFINITION  P. falciparum integration plasmid to fuse endogenous genes with tags
            using SLI.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 7486)
  AUTHORS   Birnbaum J, Flemming S, Reichard N, Soares AB, Mesen-Ramirez P, 
            Jonscher E, Bergmann B, Spielmann T
  TITLE     A genetic system to study Plasmodium falciparum protein function.
  JOURNAL   Nat Methods. 2017 Mar 13. doi: 10.1038/nmeth.4223.
  PUBMED    28288121
REFERENCE   2  (bases 1 to 7486)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nat 
            Methods. 2017 Mar 13. doi: 10.1038/nmeth.4223."
COMMENT     Sequence Label: pSLI-2xFKBP-GFP(1463000)
FEATURES             Location/Qualifiers
     source          1..7486
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     primer_bind     complement(1248..1267)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1249..1267)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1274..1291)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1274..1290)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1283..1305)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(1499..1518)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     1618..1640
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(1678..1696)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1764..1868
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1869..1937,1938..2729)
                     /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(2087..2106)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2900..3488
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3389..3408
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3642..3659
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    3776..3797
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(3812..3817,3818..3835,3836..3842)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3850..3866
                     /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     3855..3877
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     3874..3890
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3874..3890
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        3908..3926
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     3908..3925
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     CDS             4624..4944
                     /codon_start=1
                     /product="human FK506-binding protein FKBP12"
                     /label=FKBP
                     /translation="GVQVETISPGDGRTFPKRGQTCVVHYTGMLEDGKKFDSSRDRNKP
                     FKFMLGKQEVIRGWEEGVAQMSVGQRAKLTISPDYAYGATGHPGIIPPHATLVFDVELL
                     KLE"
     CDS             4951..5271
                     /codon_start=1
                     /product="human FK506-binding protein FKBP12"
                     /label=FKBP
                     /translation="GVQVETISPGDGRTFPKRGQTCVVHYTGMLEDGKKFDSSRDRNKP
                     FKFMLGKQEVIRGWEEGVAQMSVGQRAKLTISPDYAYGATGHPGIIPPHATLVFDVELL
                     KLE"
     primer_bind     complement(5356..5384)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     5979..5999
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     CDS             6049..6102
                     /codon_start=1
                     /product="2A peptide from Thosea asigna virus capsid 
                     protein"
                     /label=T2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="EGRGSLLTCGDVEENPGP"
     CDS             6097..6897
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="GPMIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQG
                     RPVLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDL
                     LSSHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEE
                     HQGLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQD
                     IALATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(6157..6176)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6767..6786
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
ORIGIN
        1 atatgtatat tttaaactag aaaaggaata actaatattt tatttattat cattcaagat
       61 ttatatttta taataataaa tacctaatag aaatatatca ggatccatgc atggttcgct
      121 aaactgcatc gttgctgtgt cccagaacat gggcatcggc aagaacgggg actacccctg
      181 gccaccgctc aggaacgaat ttagatattt ccagagaatg accacaacct cttcagtaga
      241 aggtaaacag aatctggtga ttatgggtaa gaagacctgg ttctccattc ctgagaagaa
      301 tcgaccttta aagggtagaa ttaatttagt tctcagcaga gaactcaagg aacctccaca
      361 aggagctcat tttctttcca gaagtctaga tgatgcctta aaacttactg aacaaccaga
      421 attagcaaat aaagtagaca tggtctggat agttggtggc agttctgttt ataaggaagc
      481 catgaatcac ccaggccatc ttaaactatt tgtgacaagg atcatgcaag actttgaaag
      541 tgacacgttt tttccagaaa ttgatttgga gaaatataaa cttctgccag aatacccagg
      601 tgttctctct gatgtccagg aggagaaagg cattaagtac aaatttgaag tatatgagaa
      661 gaatgattaa gcttatttaa taatagatta aaaatattat aaaaataaaa acataaacac
      721 agaaattaca aaaaaaatac atatgaattt ttttttgtaa tcttccttat aaatatagaa
      781 taatgaatca tataaaacat atcattattc atttatttac atttaaaatt attgtttcag
      841 tatctttaat ttattatgta tatataaaaa taacttacaa ttttattaat aagcaatata
      901 tgtttattaa ttcatgtttt gtaatttatg ggatagcgat tttttttact gtctgtattt
      961 ttctttttta attatgtttt aattgtattt tatttttatt attgttcttt ttatagtatt
     1021 attttaaaac aaaatgtatt ttctaagaac ttataataat aataaatata aattttaata
     1081 aaaattatat ttatctttta caatatgaac ataaagtaca acattaatat atagctttta
     1141 atatttttat tcctaatcat gtaaatctta aatttttctt tttaaacata tgttaaatat
     1201 ttatttctca ttatatataa gaacatattt attaaatcta gaattcgccc tatagtgagt
     1261 cgtattacaa ttcactggcc gtcgttttac aacgtcgtga ctgggaaaac cctggcgtta
     1321 cccaacttaa tcgccttgca gcacatcccc ctttcgccag ctggcgtaat agcgaagagg
     1381 cccgcaccga tcgcccttcc caacagttgc gcagcctgaa tggcgaatgg cgcctgatgc
     1441 ggtattttct ccttacgcat ctgtgcggta tttcacaccg catatggtgc actctcagta
     1501 caatctgctc tgatgccgca tagttaagcc agccccgaca cccgccaaca cccgctgacg
     1561 cgccctgacg ggcttgtctg ctcccggcat ccgcttacag acaagctgtg accgtctccg
     1621 ggagctgcat gtgtcagagg ttttcaccgt catcaccgaa acgcgcgaga cgaaagggcc
     1681 tcgtgatacg cctattttta taggttaatg tcatgataat aatggtttct tagacgtcag
     1741 gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt
     1801 caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa
     1861 ggaagagtat gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt
     1921 gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt
     1981 tgggtgcacg agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt
     2041 ttcgccccga agaacgtttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg
     2101 tattatcccg tattgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga
     2161 atgacttggt tgagtactca ccagtcacag aaaagcatct tacggatggc atgacagtaa
     2221 gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga
     2281 caacgatcgg aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa
     2341 ctcgccttga tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca
     2401 ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact attaactggc gaactactta
     2461 ctctagcttc ccggcaacaa ttaatagact ggatggaggc ggataaagtt gcaggaccac
     2521 ttctgcgctc ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc
     2581 gtgggtctcg cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag
     2641 ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga
     2701 taggtgcctc actgattaag cattggtaac tgtcagacca agtttactca tatatacttt
     2761 agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata
     2821 atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag
     2881 aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa
     2941 caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt
     3001 ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc
     3061 cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa
     3121 tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa
     3181 gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc
     3241 ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa
     3301 gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa
     3361 caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg
     3421 ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc
     3481 tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg
     3541 ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg
     3601 agtgagctga taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg
     3661 aagcggaaga gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat
     3721 gcagctggca cgacaggttt cccgactgga aagcgggcag tgagcgcaac gcaattaatg
     3781 tgagttagct cactcattag gcaccccagg ctttacactt tatgcttccg gctcgtatgt
     3841 tgtgtggaat tgtgagcgga taacaatttc acacaggaaa cagctatgac catgattacg
     3901 ccaagctatt taggtgacac tatagaatac tcaagctgcg gccgctaagg tatatgttgt
     3961 ttaacggttg cttcaagaat gcaacaaata aatgaaagta cttgttttaa agttatatta
     4021 gaattatcaa atttaaaaga taaattaaat gaatttaata taagacaaat tgaaatggat
     4081 gtttttgtta ccttagctca ttcaggtttt tgtttcgaaa aaattattac acctgtgaat
     4141 gaattaaatc aaataatgaa atttctagaa acatatgaaa atttttttga taataatttt
     4201 aatacattta aatcttcaag taatttaatg ctaaaaatta tttattgtat ggttccagtt
     4261 tttaatatga atatttataa ttattctatt gttaattatg ctttaaggtt atttattcaa
     4321 gataatgata aatatgaatt catttttgaa aaattaaaac aaaattatca agataaagaa
     4381 attatacata tttccaattt tcaaaatcat atgcataata ttattaatgc tttcaaaaat
     4441 acttctatct tttcagcaaa ggattctatt ttaaataatc agctagatgt tcaattgttt
     4501 gaagagataa acaaaaaaat tgaagcatat tatgaacaaa aacataaaac acctaggtca
     4561 ggattgagat caagatctgc tgctgctggt gctggtggtg ctgctagagc tgctctgcag
     4621 agaggagtac aagttgaaac aatatcacca ggagatggtc gtacatttcc aaaaagaggt
     4681 caaacttgtg ttgtacatta tactggaatg cttgaagatg gaaagaaatt tgattcatct
     4741 cgtgatagaa ataaaccatt taaatttatg ctaggtaaac aagaagtaat acgaggttgg
     4801 gaagaaggag ttgctcaaat gagtgtaggt caaagagcaa aacttactat atctccagat
     4861 tatgcttatg gtgcaactgg acatccaggt ataattccac ctcatgcaac tcttgtattt
     4921 gatgtggagc ttctaaaact agaaactaga ggtgttcagg ttgaaacaat ttcacctgga
     4981 gatggcagaa cctttcctaa aagaggacag acttgcgtag ttcattatac aggcatgcta
     5041 gaggatggta agaaatttga ttctagtcga gatagaaata agccattcaa gtttatgcta
     5101 ggtaaacagg aagtaataag aggttgggaa gagggtgtag cacagatgtc agttggacaa
     5161 agagcaaagt taacaatatc accagattat gcatacggtg caacaggcca tcctggcatc
     5221 atccctccac atgcaacttt agtattcgac gttgaattgt taaagttaga gacaacgcgt
     5281 gctagaggtg ctgctgctgg tgctggaggt gcaggtagac gtacgatgag taaaggagaa
     5341 gaacttttca ctggagttgt cccaattctt gttgaattag atggtgatgt taatgggcac
     5401 aaattttctg tcagtggaga gggtgaaggt gatgcaacat acggaaaact tacccttaaa
     5461 tttatttgca ctactggaaa actacctgtt ccatggccaa cacttgtcac tactttcgcg
     5521 tatggtcttc aatgctttgc gagataccca gatcatatga aacagcatga ctttttcaag
     5581 agtgccatgc ccgaaggtta tgtacaggaa agaactatat ttttcaaaga tgacgggaac
     5641 tacaagacac gtgctgaagt caagtttgaa ggtgataccc ttgttaatag aatcgagtta
     5701 aaaggtattg attttaaaga agatggaaac attcttggac acaaattgga atacaactat
     5761 aactcacaca atgtatacat catggcagac aaacaaaaga atggaatcaa agttaacttc
     5821 aaaattagac acaacattga agatggaagc gttcaactag cagaccatta tcaacaaaat
     5881 actccaattg gcgatggccc tgtcctttta ccagacaacc attacctgtc cacacaatct
     5941 gccctttcga aagatcccaa cgaaaagaga gaccacatgg tccttcttga gtttgtaaca
     6001 gctgctggga ttacacatgg catggatgag ctctacaaag tcgacggaga aggaagagga
     6061 agtttattaa catgtggaga tgtagaagaa aatccaggac caatgattga acaagatgga
     6121 ttgcacgcag gttctccggc cgcttgggtg gagaggctat tcggctatga ctgggcacaa
     6181 cagacaatcg gctgctctga tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt
     6241 ctttttgtca agaccgacct gtccggtgcc ctgaatgaac tgcaggacga ggcagcgcgg
     6301 ctatcgtggc tggccacgac gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa
     6361 gcgggaaggg actggctgct attgggcgaa gtgccggggc aggatctcct gtcatctcac
     6421 cttgctcctg ccgagaaagt atccatcatg gctgatgcaa tgcggcggct gcatacgctt
     6481 gatccggcta cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact
     6541 cggatggaag ccggtcttgt cgatcaggat gatctggacg aagagcatca ggggctcgcg
     6601 ccagccgaac tgttcgccag gctcaaggcg cgcatgcccg acggcgagga tctcgtcgtg
     6661 acccatggcg atgcctgctt gccgaatatc atggtggaaa atggccgctt ttctggattc
     6721 atcgactgtg gccggctggg tgtggcggac cgctatcagg acatagcgtt ggctacccgt
     6781 gatattgctg aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc
     6841 gccgctcccg attcgcagcg catcgccttc tatcgccttc ttgacgagtt cttctaactc
     6901 gagggatatg gcagcttaat gttcgttttt cttatttata tatttatacc aattgattgt
     6961 atttataact gtaaaaatgt gtatgttgtg tgcatatttt tttttgtgca tgcacatgca
     7021 tgtaaatagc taaaattatg aacattttat tttttgttca gaaaaaaaaa actttacaca
     7081 cataaaatgg ctagtatgaa tagccatatt ttatataaat taaatcctat gaatttatga
     7141 ccatattaaa aatttagata tttatggaac ataatatgtt tgaaacaata agacaaaatt
     7201 attattatta ttattatttt tactgttata attatgttgt ctcttcaatg attcataaat
     7261 agttggactt gatttttaaa atgtttataa tatgattagc atagttaaat aaaaaaagtt
     7321 gaaaaattaa aaaaaaacat ataaacacaa atgatgtttt ttccttcaat ttcgatatca
     7381 atttatagaa acaaaatata tacttgtata attttatttt tttatataaa tcattacata
     7441 tataattata caatattttt tctaagagat aattatatat taatat
//
