LOCUS Exported 9049 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION P. falciparum integration plasmid to fuse endogenous genes with tags (double 2xFKBP tag) using SLI. ACCESSION . VERSION . KEYWORDS pSLI-sandwich(0807600) SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 9049) 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 9049) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..9049 /organism="synthetic DNA construct" /mol_type="other DNA" promoter complement(2448..2465) /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" 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 3067..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 5010..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" CDS 6032..6352 /codon_start=1 /product="human FK506-binding protein FKBP12" /label=FKBP /translation="GVQVETISPGDGRTFPKRGQTCVVHYTGMLEDGKKFDSSRDRNKP FKFMLGKQEVIRGWEEGVAQMSVGQRAKLTISPDYAYGATGHPGIIPPHATLVFDVELL KLE" CDS 6359..6679 /codon_start=1 /product="human FK506-binding protein FKBP12" /label=FKBP /translation="GVQVETISPGDGRTFPKRGQTCVVHYTGMLEDGKKFDSSRDRNKP FKFMLGKQEVIRGWEEGVAQMSVGQRAKLTISPDYAYGATGHPGIIPPHATLVFDVELL KLE" primer_bind complement(6764..6792) /label=GFP-R /note="GFP, reverse primer. Does NOT anneal to EGFP" primer_bind 7387..7407 /label=GFP-F /note="GFP, forward primer. Does NOT anneal to EGFP" CDS 7496..7816 /codon_start=1 /product="human FK506-binding protein FKBP12" /label=FKBP /translation="GVQVETISPGDGRTFPKRGQTCVVHYTGMLEDGKKFDSSRDRNKP FKFMLGKQEVIRGWEEGVAQMSVGQRAKLTISPDYAYGATGHPGIIPPHATLVFDVELL KLE" CDS 7862..8182 /codon_start=1 /product="human FK506-binding protein FKBP12" /label=FKBP /translation="GVQVETISPGDGRTFPKRGQTCVVHYTGMLEDGKKFDSSRDRNKP FKFMLGKQEVIRGWEEGVAQMSVGQRAKLTISPDYAYGATGHPGIIPPHATLVFDVELL KLE" CDS 8195..8248 /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 8249..9043 /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(8303..8322) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 8913..8932 /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 aaggggtggt tatagtatga 5161 gaaatgatca acgaaatgat caacgaaatg atcaacgaaa tgatcaacga aatgatcata 5221 ggaatgacca tagaaatgat cacagaaatg atcacagaaa tgatcacaga aatgatcaca 5281 gaaatgatca cagaaatgat catagaaatg atcatagaaa tgatcaaaga aatgatcata 5341 gaaatagtat gaggaatgac catagaaata gtatgaggaa tgaccataga aatagtatga 5401 ggaatgatca acgaaatagt atgatgaatg atcaacgaaa tagtatgatg aacgatcaac 5461 gaaatagtat gatgaacgat caacgaaatg ttaaacgaag aagagaagaa tcgcatgaaa 5521 tgaatagaag aggagatata tattcaaata aaagatatga caaaagagga agaatagatt 5581 atgatatgaa attaaaaatg gatgagaaaa aaaatgagaa aaatattaag aatcattcag 5641 aaatgcagga tccatatgca tcaacaaaag atacctctag aaattttgat atgaataatt 5701 ttacaattag ttctgaaagg aaatcatgga aattacctcc tccacctcct aacattcctc 5761 catcctatga taggtcatct caacgaagaa cggtaaaatg aaagaaatta ataatatata 5821 tgtatgtata tatatatata tatatatata tatatatata tataatatat atatgtgttt 5881 gcatttattt atatgtttgt atttatttat gtgcttcatt ttttagagca atttttacaa 5941 taatgacgat tcttcaatgc ctaggtcagg attgagatca agatctgctg ctgctggtgc 6001 tggtggtgct gctagagctg ctctgcagag aggagtacaa gttgaaacaa tatcaccagg 6061 agatggtcgt acatttccaa aaagaggtca aacttgtgtt gtacattata ctggaatgct 6121 tgaagatgga aagaaatttg attcatctcg tgatagaaat aaaccattta aatttatgct 6181 aggtaaacaa gaagtaatac gaggttggga agaaggagtt gctcaaatga gtgtaggtca 6241 aagagcaaaa cttactatat ctccagatta tgcttatggt gcaactggac atccaggtat 6301 aattccacct catgcaactc ttgtatttga tgtggagctt ctaaaactag aaactagagg 6361 tgttcaggtt gaaacaattt cacctggaga tggcagaacc tttcctaaaa gaggacagac 6421 ttgcgtagtt cattatacag gcatgctaga ggatggtaag aaatttgatt ctagtcgaga 6481 tagaaataag ccattcaagt ttatgctagg taaacaggaa gtaataagag gttgggaaga 6541 gggtgtagca cagatgtcag ttggacaaag agcaaagtta acaatatcac cagattatgc 6601 atacggtgca acaggccatc ctggcatcat ccctccacat gcaactttag tattcgacgt 6661 tgaattgtta aagttagaga caacgcgtgc tagaggtgct gctgctggtg ctggaggtgc 6721 aggtagacgt acgatgagta aaggagaaga acttttcact ggagttgtcc caattcttgt 6781 tgaattagat ggtgatgtta atgggcacaa attttctgtc agtggagagg gtgaaggtga 6841 tgcaacatac ggaaaactta cccttaaatt tatttgcact actggaaaac tacctgttcc 6901 atggccaaca cttgtcacta ctttcgcgta tggtcttcaa tgctttgcga gatacccaga 6961 tcatatgaaa cagcatgact ttttcaagag tgccatgccc gaaggttatg tacaggaaag 7021 aactatattt ttcaaagatg acgggaacta caagacacgt gctgaagtca agtttgaagg 7081 tgataccctt gttaatagaa tcgagttaaa aggtattgat tttaaagaag atggaaacat 7141 tcttggacac aaattggaat acaactataa ctcacacaat gtatacatca tggcagacaa 7201 acaaaagaat ggaatcaaag ttaacttcaa aattagacac aacattgaag atggaagcgt 7261 tcaactagca gaccattatc aacaaaatac tccaattggc gatggccctg tccttttacc 7321 agacaaccat tacctgtcca cacaatctgc cctttcgaaa gatcccaacg aaaagagaga 7381 ccacatggtc cttcttgagt ttgtaacagc tgctgggatt acacatggca tggatgagct 7441 ctacaaagtc gacgccaggg gagcagccgc aggagcaggg ggggcaggaa ggcgtggtgt 7501 tcaggtcgag actattagcc ctggagatgg acgcacgttt cctaagcgtg gacagacatg 7561 cgtagttcac tacacaggta tgttggagga cggtaaaaag ttcgacagct cacgcgaccg 7621 caataaacct ttcaagttta tgcttggcaa gcaggaggtt attcgtggat gggaggaggg 7681 tgtagcacag atgtctgttg gacagcgtgc taagttgaca atttcacctg actatgctta 7741 tggcgctacg ggccatcccg ggatcattcc gccacatgcg actctggtat tcgacgttga 7801 attattaaag ttagagacag ctagaggggc cgctgcaggt gctggtggag ctggaagacg 7861 tggagtacaa gtagagacta tctctccagg tgacggtcgc actttcccaa agcgtggcca 7921 aacctgtgtt gtacattaca ctggtatgct ggaggatggg aaaaagttcg attccagtcg 7981 cgaccgtaac aaaccgttca aattcatgtt gggaaagcag gaagtgatcc gcgggtggga 8041 ggaaggcgtg gcgcaaatga gcgtcggtca gcgggctaaa ttgaccattt cccctgacta 8101 cgcgtatggg gctactgggc acccagggat tattccgcct cacgctacac ttgtgtttga 8161 tgtcgaactt ttgaaactgg aaactgtcga cggagaagga agaggaagtt tattaacatg 8221 tggagatgta gaagaaaatc caggaccaat gattgaacaa gatggattgc acgcaggttc 8281 tccggccgct tgggtggaga ggctattcgg ctatgactgg gcacaacaga caatcggctg 8341 ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac 8401 cgacctgtcc ggtgccctga atgaactgca ggacgaggca gcgcggctat cgtggctggc 8461 cacgacgggc gttccttgcg cagctgtgct cgacgttgtc actgaagcgg gaagggactg 8521 gctgctattg ggcgaagtgc cggggcagga tctcctgtca tctcaccttg ctcctgccga 8581 gaaagtatcc atcatggctg atgcaatgcg gcggctgcat acgcttgatc cggctacctg 8641 cccattcgac caccaagcga aacatcgcat cgagcgagca cgtactcgga tggaagccgg 8701 tcttgtcgat caggatgatc tggacgaaga gcatcagggg ctcgcgccag ccgaactgtt 8761 cgccaggctc aaggcgcgca tgcccgacgg cgaggatctc gtcgtgaccc atggcgatgc 8821 ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct ggattcatcg actgtggccg 8881 gctgggtgtg gcggaccgct atcaggacat agcgttggct acccgtgata ttgctgaaga 8941 gcttggcggc gaatgggctg accgcttcct cgtgctttac ggtatcgccg ctcccgattc 9001 gcagcgcatc gccttctatc gccttcttga cgagttcttc taactcgag //