LOCUS       sequence-166790-        7260 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  P. falciparum integration plasmid to disrupt genes using SLI.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7260)
  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 7260)
  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: "Nat 
            Methods. 2017 Mar 13. doi: 10.1038/nmeth.4223."
COMMENT     Sequence Label: pSLI-TGD(1463000)
FEATURES             Location/Qualifiers
     source          1..7260
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(2472..2489)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2472..2488)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2481..2503)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(2697..2716)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2816..2838
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2876..2894)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2962..3066
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3067..3135,3136..3927)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMPTTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(3285..3304)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4098..4686
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4587..4606
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4840..4857
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    4974..4995
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(5010..5015,5016..5033,5034..5040)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5048..5064
                     /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     5053..5075
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     5072..5088
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5072..5088
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        5106..5124
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     5106..5123
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(5713..5741)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     6336..6356
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     CDS             6406..6459
                     /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             6454..7254
                     /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(6514..6533)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     7124..7143
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
ORIGIN
        1 ggatatggca gcttaatgtt cgtttttctt atttatatat ttataccaat tgattgtatt
       61 tataactgta aaaatgtgta tgttgtgtgc atattttttt ttgtgcatgc acatgcatgt
      121 aaatagctaa aattatgaac attttatttt ttgttcagaa aaaaaaaact ttacacacat
      181 aaaatggcta gtatgaatag ccatatttta tataaattaa atcctatgaa tttatgacca
      241 tattaaaaat ttagatattt atggaacata atatgtttga aacaataaga caaaattatt
      301 attattatta ttatttttac tgttataatt atgtgtctcc ttcaatgatt cataaatagt
      361 tggacttgat ttttaaaatg tttataatat gattagcata gttaaataaa aaaagttgaa
      421 aaattaaaaa aaaacatata aacacaaatg atggtttttc cttcaatttc gatatcaatt
      481 tatagaaaca aaatatatac ttgtataatt ttattttttt atataaatca ttacatatat
      541 aattatacaa tattttttct aagagataat tatatattaa tatatataaa aaaaggtgtt
      601 tttttttttt ttttttattt ttatttttat tttatggtaa tattttattt tccttatttt
      661 ataaattata ttagtttata tgtgattaat tttatatatt atcaatttat atatttttaa
      721 atgcttactt aattatcttt tttttttttt tttttttttt ttcccctctt tttatattaa
      781 tttatttttg aaaaaattga tatatatata tatatataat atatatatat acatgtagta
      841 gtattaaaca atgtataata tatataaata atatatttat atatttcatt tcaattttaa
      901 ttttttttgg tttttttttt ttttcttttt gtcatattta aaaaaaatta tattcatata
      961 agttatgcat tttttataaa cattattcaa tatatgtata atataatata tatatatata
     1021 ttaatgtatt attccaatgt gcatgataaa agaaaaaaat aatatttata aaaaaaaaga
     1081 aaaataaaac aaaaaaagaa aaaaaaaaaa aaaaaaaaaa aaatacaaaa ataaataata
     1141 taatttataa ttatatattc ttgtcacaat aaaaatatat atatatatat atatatttat
     1201 aatatgtata ttttaaacta gaaaaggaat aactaatatt ttatttatta tcattcaaga
     1261 tttatatttt ataataataa atacctaata gaaatatatc aggatccatg catggttcgc
     1321 taaactgcat cgtcgctgtg tcccagaaca tgggcatcgg caagaacggg gactacccct
     1381 ggccaccgct caggaacgaa tttagatatt tccagagaat gaccacaacc tcttcagtag
     1441 aaggtaaaca gaatctggtg attatgggta agaagacctg gttctccatt cctgagaaga
     1501 atcgaccttt aaagggtaga attaatttag ttctcagcag agaactcaag gaacctccac
     1561 aaggagctca ttttctttcc agaagtctag atgatgcctt aaaacttact gaacaaccag
     1621 aattagcaaa taaagtagac atggtctgga tagttggtgg cagttctgtt tataaggaag
     1681 ccatgaatca cccaggccat cttaaactat ttgtgacaag gatcatgcaa gactttgaaa
     1741 gtgacacgtt ttttccagaa attgatttgg agaaatataa acttctgcca gaatacccag
     1801 gtgttctctc tgatgtccag gaggagaaag gcattaagta caaatttgaa gtatatgaga
     1861 agaatgatta agcttattta ataatagatt aaaaatatta taaaaataaa aacataaaca
     1921 cagaaattac aaaaaaaata catatgaatt ttttttttgt aatcttcctt ataaatatag
     1981 aataatgaat catataaaac atatcattat tcatttattt acatttaaaa ttattgtttc
     2041 agtatcttta atttattatg tatatataaa aataacttac aattttatta ataaacaata
     2101 tatgtttatt aattcatgtt ttgtaattta tgggatagcg atttttttta ctgtctgtat
     2161 ttttcttttt taattatgtt ttaattgtat tttattttta ttattgttct ttttatagta
     2221 ttattttaaa acaaaatgta ttttctaaga acttataata ataataaata taaattttaa
     2281 taaaaattat atttatcttt tacaatatga acataaagta caacattaat atatagcttt
     2341 taatattttt attcctaatc atgtaaatct taaatttttc tttttaaaca tatgttaaat
     2401 atttatttct cattatatat aagaacatat ttattaaatc tagaattcta tagtgagtcg
     2461 tattacaatt cactggccgt cgttttacaa cgtcgtgact gggaaaaccc tggcgttacc
     2521 caacttaatc gccttgcagc acatccccct ttcgccagct ggcgtaatag cgaagaggcc
     2581 cgcaccgatc gcccttccca acagttgcgc agcctgaatg gcgaatggcg cctgatgcgg
     2641 tattttctcc ttacgcatct gtgcggtatt tcacaccgca tatggtgcac tctcagtaca
     2701 atctgctctg atgccgcata gttaagccag ccccgacacc cgccaacacc cgctgacgcg
     2761 ccctgacggg cttgtctgct cccggcatcc gcttacagac aagctgtgac cgtctccggg
     2821 agctgcatgt gtcagaggtt ttcaccgtca tcaccgaaac gcgcgagacg aaagggcctc
     2881 gtgatacgcc tatttttata ggttaatgtc atgataataa tggtttctta gacgtcaggt
     2941 ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca
     3001 aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg
     3061 aagagtatga gtattcaaca tttccgtgtc gcccttattc ccttttttgc ggcattttgc
     3121 cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga agatcagttg
     3181 ggtgcacgag tgggttacat cgaactggat ctcaacagcg gtaagatcct tgagagtttt
     3241 cgccccgaag aacgttttcc aatgatgagc acttttaaag ttctgctatg tggcgcggta
     3301 ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat
     3361 gacttggttg agtactcacc agtcacagaa aagcatctta cggatggcat gacagtaaga
     3421 gaattatgca gtgctgccat aaccatgagt gataacactg cggccaactt acttctgaca
     3481 acgatcggag gaccgaagga gctaaccgct tttttgcaca acatggggga tcatgtaact
     3541 cgccttgatc gttgggaacc ggagctgaat gaagccatac caaacgacga gcgtgacacc
     3601 acgatgcctg tagcaatgcc aacaacgttg cgcaaactat taactggcga actacttact
     3661 ctagcttccc ggcaacaatt aatagactgg atggaggcgg ataaagttgc aggaccactt
     3721 ctgcgctcgg cccttccggc tggctggttt attgctgata aatctggagc cggtgagcgt
     3781 gggtctcgcg gtatcattgc agcactgggg ccagatggta agccctcccg tatcgtagtt
     3841 atctacacga cggggagtca ggcaactatg gatgaacgaa atagacagat cgctgagata
     3901 ggtgcctcac tgattaagca ttggtaactg tcagaccaag tttactcata tatactttag
     3961 attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat
     4021 ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa
     4081 aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca
     4141 aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt
     4201 ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgtccttct agtgtagccg
     4261 tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc
     4321 ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga
     4381 cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc
     4441 agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc
     4501 gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca
     4561 ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg
     4621 tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta
     4681 tcgaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct
     4741 cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac cgcctttgag
     4801 tgagctgata ccgctcgccg cagccgaacg accgagcgca gcgagtcagt gagcgaggaa
     4861 gcggaagagc gcccaatacg caaaccgcct ctccccgcgc gttggccgat tcattaatgc
     4921 agctggcacg acaggtttcc cgactggaaa gcgggcagtg agcgcaacgc aattaatgtg
     4981 agttagctca ctcattaggc accccaggct ttacacttta tgcttccggc tcgtatgttg
     5041 tgtggaattg tgagcggata acaatttcac acaggaaaca gctatgacca tgattacgcc
     5101 aagctattta ggtgacacta tagaatactc gcggccgcta agcgtgaaat aattacgaat
     5161 accagaaaga taaaattaca tgagaatttt ttagaggatt gtaatttaaa agatatattt
     5221 tataatggat tggaaacagt aaattgtgga tataatgatt atgtaaatat gaaaacatgg
     5281 ctatttgttt tatcagaaaa aatgtttcgt acggattatg aagagctagc atatgtagca
     5341 tgtgatgctt taaaagtatt attatgtaat agtaaatttt taaatgttat aatttttaaa
     5401 ttattaagaa aaaaattaga aaaagaaatt ttaaagatta tgggtatatg ttgtttaacg
     5461 gttgcttcaa gaatgcaaca aataaatgaa agtacttgtt ttaaagttat attagaatta
     5521 tcaaatttaa aagataaatt aaatgaattt aatataagac aaattgaaat ggatgttttt
     5581 gttaccttag ctcattcagg tttttgtttc gaaaaaatta ttacacctgt gacgcgtgct
     5641 agaggtgctg ctgctggtgc tggaggtgca ggtagacgta cgatgagtaa aggagaagaa
     5701 cttttcactg gagttgtccc aattcttgtt gaattagatg gtgatgttaa tgggcacaaa
     5761 ttttctgtca gtggagaggg tgaaggtgat gcaacatacg gaaaacttac ccttaaattt
     5821 atttgcacta ctggaaaact acctgttcca tggccaacac ttgtcactac tttcgcgtat
     5881 ggtcttcaat gctttgcgag atacccagat catatgaaac agcatgactt tttcaagagt
     5941 gccatgcccg aaggttatgt acaggaaaga actatatttt tcaaagatga cgggaactac
     6001 aagacacgtg ctgaagtcaa gtttgaaggt gatacccttg ttaatagaat cgagttaaaa
     6061 ggtattgatt ttaaagaaga tggaaacatt cttggacaca aattggaata caactataac
     6121 tcacacaatg tatacatcat ggcagacaaa caaaagaatg gaatcaaagt taacttcaaa
     6181 attagacaca acattgaaga tggaagcgtt caactagcag accattatca acaaaatact
     6241 ccaattggcg atggccctgt ccttttacca gacaaccatt acctgtccac acaatctgcc
     6301 ctttcgaaag atcccaacga aaagagagac cacatggtcc ttcttgagtt tgtaacagct
     6361 gctgggatta cacatggcat ggatgagctc tacaaagtcg acggagaagg aagaggaagt
     6421 ttattaacat gtggagatgt agaagaaaat ccaggaccaa tgattgaaca agatggattg
     6481 cacgcaggtt ctccggccgc ttgggtggag aggctattcg gctatgactg ggcacaacag
     6541 acaatcggct gctctgatgc cgccgtgttc cggctgtcag cgcaggggcg cccggttctt
     6601 tttgtcaaga ccgacctgtc cggtgccctg aatgaactgc aggacgaggc agcgcggcta
     6661 tcgtggctgg ccacgacggg cgttccttgc gcagctgtgc tcgacgttgt cactgaagcg
     6721 ggaagggact ggctgctatt gggcgaagtg ccggggcagg atctcctgtc atctcacctt
     6781 gctcctgccg agaaagtatc catcatggct gatgcaatgc ggcggctgca tacgcttgat
     6841 ccggctacct gcccattcga ccaccaagcg aaacatcgca tcgagcgagc acgtactcgg
     6901 atggaagccg gtcttgtcga tcaggatgat ctggacgaag agcatcaggg gctcgcgcca
     6961 gccgaactgt tcgccaggct caaggcgcgc atgcccgacg gcgaggatct cgtcgtgacc
     7021 catggcgatg cctgcttgcc gaatatcatg gtggaaaatg gccgcttttc tggattcatc
     7081 gactgtggcc ggctgggtgt ggcggaccgc tatcaggaca tagcgttggc tacccgtgat
     7141 attgctgaag agcttggcgg cgaatgggct gaccgcttcc tcgtgcttta cggtatcgcc
     7201 gctcccgatt cgcagcgcat cgccttctat cgccttcttg acgagttctt ctaactcgag
//
