LOCUS Exported 4649 bp ds-DNA linear SYN 04-JUN-2021 DEFINITION P. falciparum integration plasmid to disrupt genes using SLI. ACCESSION . VERSION . KEYWORDS pSLI-TGD(1463000) SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 4649) 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 4649) AUTHORS . TITLE Direct Submission JOURNAL Exported Jun 4, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..4649 /organism="synthetic DNA construct" /mol_type="other DNA" promoter 26..130 /gene="bla" /label=AmpR promoter CDS 131..991 /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(349..368) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 1162..1750 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 1651..1670 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 1904..1921 /label=L4440 /note="L4440 vector, forward primer" protein_bind 2038..2059 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." promoter 2074..2104 /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 2112..2128 /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 2117..2139 /label=M13/pUC Reverse /note="In lacZ gene" primer_bind 2136..2152 /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind 2136..2152 /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" promoter 2170..2188 /label=SP6 promoter /note="promoter for bacteriophage SP6 RNA polymerase" primer_bind 2170..2187 /label=SP6 /note="SP6 promoter, forward primer" primer_bind complement(2782..2810) /label=GFP-R /note="GFP, reverse primer. Does NOT anneal to EGFP" primer_bind 3405..3425 /label=GFP-F /note="GFP, forward primer. Does NOT anneal to EGFP" CDS 3475..3528 /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 3529..4323 /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="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF" primer_bind complement(3583..3602) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 4193..4212 /label=Neo-F /note="Neomycin resistance gene, forward primer" ORIGIN 1 aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca 61 ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa 121 aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt ttgcggcatt 181 ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg ctgaagatca 241 gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga tccttgagag 301 ttttcgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc tatgtggcgc 361 ggtattatcc cgtattgacg ccgggcaaga gcaactcggt cgccgcatac actattctca 421 gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg gcatgacagt 481 aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca acttacttct 541 gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg gggatcatgt 601 aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg acgagcgtga 661 caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg gcgaactact 721 tactctagct tcccggcaac aattaataga ctggatggag gcggataaag ttgcaggacc 781 acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg gagccggtga 841 gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct cccgtatcgt 901 agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac agatcgctga 961 gataggtgcc tcactgatta agcattggta actgtcagac caagtttact catatatact 1021 ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga 1081 taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt 1141 agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca 1201 aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct 1261 ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc ttctagtgta 1321 gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct 1381 aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc 1441 aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca 1501 gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga 1561 aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg 1621 aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt 1681 cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag 1741 cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt 1801 tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgcctt 1861 tgagtgagct gataccgctc gccgcagccg aacgaccgag cgcagcgagt cagtgagcga 1921 ggaagcggaa gagcgcccaa tacgcaaacc gcctctcccc gcgcgttggc cgattcatta 1981 atgcagctgg cacgacaggt ttcccgactg gaaagcgggc agtgagcgca acgcaattaa 2041 tgtgagttag ctcactcatt aggcacccca ggctttacac tttatgcttc cggctcgtat 2101 gttgtgtgga attgtgagcg gataacaatt tcacacagga aacagctatg accatgatta 2161 cgccaagcta tttaggtgac actatagaat actcaagctg cggccgctaa gcgtgaaata 2221 attacgaata ccagaaagat aaaattacat gagaattttt tagaggattg taatttaaaa 2281 gatatatttt ataatggatt ggaaacagta aattgtggat ataatgatta tgtaaatatg 2341 aaaacatggc tatttgtttt atcagaaaaa atgtttcgta cggattatga agagctagca 2401 tatgtagcat gtgatgcttt aaaagtatta ttatgtaata gtaaattttt aaatgttata 2461 atttttaaat tattaagaaa aaaattagaa aaagaaattt taaagattat gggtatatgt 2521 tgtttaacgg ttgcttcaag aatgcaacaa ataaatgaaa gtacttgttt taaagttata 2581 ttagaattat caaatttaaa agataaatta aatgaattta atataagaca aattgaaatg 2641 gatgtttttg ttaccttagc tcattcaggt ttttgtttcg aaaaaattat tacacctgtg 2701 acgcgtgcta gaggtgctgc tgctggtgct ggaggtgcag gtagacgtac gatgagtaaa 2761 ggagaagaac ttttcactgg agttgtccca attcttgttg aattagatgg tgatgttaat 2821 gggcacaaat tttctgtcag tggagagggt gaaggtgatg caacatacgg aaaacttacc 2881 cttaaattta tttgcactac tggaaaacta cctgttccat ggccaacact tgtcactact 2941 ttcgcgtatg gtcttcaatg ctttgcgaga tacccagatc atatgaaaca gcatgacttt 3001 ttcaagagtg ccatgcccga aggttatgta caggaaagaa ctatattttt caaagatgac 3061 gggaactaca agacacgtgc tgaagtcaag tttgaaggtg atacccttgt taatagaatc 3121 gagttaaaag gtattgattt taaagaagat ggaaacattc ttggacacaa attggaatac 3181 aactataact cacacaatgt atacatcatg gcagacaaac aaaagaatgg aatcaaagtt 3241 aacttcaaaa ttagacacaa cattgaagat ggaagcgttc aactagcaga ccattatcaa 3301 caaaatactc caattggcga tggccctgtc cttttaccag acaaccatta cctgtccaca 3361 caatctgccc tttcgaaaga tcccaacgaa aagagagacc acatggtcct tcttgagttt 3421 gtaacagctg ctgggattac acatggcatg gatgagctct acaaagtcga cggagaagga 3481 agaggaagtt tattaacatg tggagatgta gaagaaaatc caggaccaat gattgaacaa 3541 gatggattgc acgcaggttc tccggccgct tgggtggaga ggctattcgg ctatgactgg 3601 gcacaacaga caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc 3661 ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga atgaactgca ggacgaggca 3721 gcgcggctat cgtggctggc cacgacgggc gttccttgcg cagctgtgct cgacgttgtc 3781 actgaagcgg gaagggactg gctgctattg ggcgaagtgc cggggcagga tctcctgtca 3841 tctcaccttg ctcctgccga gaaagtatcc atcatggctg atgcaatgcg gcggctgcat 3901 acgcttgatc cggctacctg cccattcgac caccaagcga aacatcgcat cgagcgagca 3961 cgtactcgga tggaagccgg tcttgtcgat caggatgatc tggacgaaga gcatcagggg 4021 ctcgcgccag ccgaactgtt cgccaggctc aaggcgcgca tgcccgacgg cgaggatctc 4081 gtcgtgaccc atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct 4141 ggattcatcg actgtggccg gctgggtgtg gcggaccgct atcaggacat agcgttggct 4201 acccgtgata ttgctgaaga gcttggcggc gaatgggctg accgcttcct cgtgctttac 4261 ggtatcgccg ctcccgattc gcagcgcatc gccttctatc gccttcttga cgagttcttc 4321 taactcgagg gatatggcag cttaatgttc gtttttctta tttatatatt tataccaatt 4381 gattgtattt ataactgtaa aaatgtgtat gttgtgtgca tatttttttt tgtgcatgca 4441 catgcatgta aatagctaaa attatgaaca ttttattttt tgttcagaaa aaaaaaactt 4501 tacacacata aaatggctag tatgaatagc catattttat ataaattaaa tcctatgaat 4561 ttatgaccat attaaaaatt tagatattta tggaacataa tatgtttgaa acaataagac 4621 aaaattatta ttattattat tatttttac //