LOCUS Exported 5844 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Work as template for amplification of Streptococcus pneumoniae adapted lacI gene, which is flanked by up- and downstream sequence for integration in prsA locus in S. pneumoniae. ACCESSION . VERSION . KEYWORDS pPEPY-PF6-lacI SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 5844) AUTHORS Liu X, Gallay C, Kjos M, Domenech A, Slager J, van Kessel SP, Knoops K, Sorg RA, Zhang JR, Veening JW TITLE High-throughput CRISPRi phenotyping identifies new essential genes in Streptococcus pneumoniae. JOURNAL Mol Syst Biol. 2017 May 10;13(5):931. doi: 10.15252/msb.20167449. PUBMED 28490437 REFERENCE 2 (bases 1 to 5844) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..5844 /organism="synthetic DNA construct" /mol_type="other DNA" rep_origin 49..637 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 538..557 /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(2033..2566) /codon_start=1 /gene="aacC1" /product="gentamycin acetyltransferase" /label=GmR /note="confers resistance to gentamycin" /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR EEVMHFDIDPSTAT" CDS 4469..5263 /codon_start=1 /gene="aphA-3" /product="aminoglycoside phosphotransferase" /label=KanR /note="confers resistance to kanamycin" /translation="MAKMRISPELKKLIEKYRCVKDTEGMSPAKVYKLVGENENLYLKM TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEEYED EQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA FCVRSIREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF" ORIGIN 1 ctttagtaag gtttggtacc accccgtaga aaagatcaaa ggatcttctt gagatccttt 61 ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg 121 tttgccggat caagagctac caactctttt tccgaaggta actggcttca gcagagcgca 181 gataccaaat actgtccttc tagtgtagcc gtagttaggc caccacttca agaactctgt 241 agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga 301 taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc 361 gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact 421 gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga 481 caggtatcca gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg 541 aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt 601 tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt 661 acggttcccg gccttttgct ggccttttgc tcacatgttc tttcctgcgt tatcccgcgg 721 ccgcataaaa acattcatca taacccccag aacttaaata acaattttta ttcaagatac 781 atactcctag aataaacttt atatgaaatt ctcatttttg tttttacaat tctccttagt 841 taaatcttgt ttaatatatg ttttacatat agtatttagc gccacatagt actgaactct 901 ctccaaaaac ggttattcct ctttgaatag ggcgttatca caagaaaagt atctccacgt 961 ttcaacttca tatggctcaa aaacaatcaa ttgatgctaa aacctgtacc tagatgtttc 1021 ggttcataaa accatgaaac tgtaaaagtg gatgaaattg atagcgatag tcaaatcaag 1081 aggcatcata actctaaaaa gtcacaatat ataagttcat cctcggaaaa atatcattct 1141 aattgttgaa atgcctacat gaaaagaaac gtcaaatgct catgaaacaa cgaatacagg 1201 tatcaaaact atgacaaaac aagtccctaa atttactaaa gacactgctc aactttacac 1261 tttctcgagt tactggaggg atccggcatt aattttcctc cttatttatt tagatctaca 1321 aacacatcgc tcctgtttac gtcattgcta tcgtggatag taattatttc tcagatgacc 1381 tggcttttca tagctttagt atgcgcgaag acacaacagg tgaggtattg gcgattacca 1441 acaatggaca ggaaaaccat ctggttaaga tggcattctt ggaattaaaa aatacagaga 1501 aaccagcaaa gacaaggttc gcaagccatg gttggagttt ttcggcaaca agccctttac 1561 ccagcaaccg caacgagcca ttacccaagc aaatcaactg ctggactaca agagctggtc 1621 cgaggaggac aggaaaatgt ttagtcaact acatatgcga gaagaacaag tcttgttagc 1681 acaggactat gccttggaaa ctgctagggc tgaagacctt gaacaaggac tagagcgtgg 1741 gaaagttgaa ggaagggcag aaaggaaact ttttaccttc ctagacatag tacgccaagg 1801 tcttctgact tctgaggttg ccagccagca attaggtatg tcagtatctg aatttgaggc 1861 actgttgtaa aatggctcca taatatccat agtgagtaaa tcccctatgg atattatgga 1921 gcctattttg tgtagtgtac agttaccata cgtaggtgtc cacttaaaaa aaaaccgcgc 1981 cctgtcaggg gcgcggtttt tttttctcga gttactggag ggatccttat tattaggtgg 2041 cggtacttgg gtcgatatca aagtgcatca cttcttcccg tatgcccaac tttgtataga 2101 gagccactgc gggatcgtca ccgtaatctg cttgcacgta gatcacataa gcaccaagcg 2161 cgttggcctc atgcttgagg agattgatga gcgcggtggc aatgccctgc ctccggtgct 2221 cgccggagac tgcgagatca tagatataga tttcactacg cggctgctca aacttgggca 2281 gaacgtaagc cgcgagagcg ccaacaaccg cttcttggtc gaaggcagca agcgcgatga 2341 atgtcttact acggagcaag ttcccgaggt aatcggagtc cggctgatgt tgggagtagg 2401 tggctacgtc tccgaactca cgaccgaaaa gatcaagagc agcccgcatg gatttgactt 2461 ggtcagggcc gagcctacat gtgcgaatga tgcccatact tgagccacct aactttgttt 2521 tagggcgact gccctgctgc gtaacatcgt tgctgctgcg taacattaat tttcctccta 2581 tttagatcct tattattatt gtccactttc caaacgagag acctgacgtg caagctgcat 2641 aagactatct gccaaggcac gaggtgaggc ggtttgagta tttggtgcta atgtggtctt 2701 acgcttaaca agagatacag gtaaaagctg atttcccttg acagcctgac cctgagaaag 2761 ttgcaataaa cgatcaacac ttgtctgtcc aagcaaacga aaatcttgct tgatagttgt 2821 caaaggtgga atgtaacaac ttgaatcttc tgtatcatcg taaccgacaa cactaatatc 2881 agctccaaca cgcaaacctg attcggtaat tgcacgcata gctccaaggg ccatttggtc 2941 attggcaacc aacattgcag ttggtacaat accttcattc aacatttgca ttgtctgttg 3001 aaatcctgac attgcagacc aatcaccctc acgttcagcg ataggctgaa tttgattacg 3061 agttaagtat ttgtgccaac cagctaaacg aagacgggct gaaacagaac taagtggtcc 3121 tgctaaaaga gcaatctgtt ggtgaccaag agctaccaaa tgttcgactc ccaaacgagt 3181 accatcttca tgactaaaga tgattgaatt gataggtgtt tgatctgaaa catccaaaaa 3241 taatgctggt acattggtac atgcagcttc gacagcaatg gcatcttggt catccaatgg 3301 gtaattgata attaaaccag atacacgttg agccaataaa ttgtggacgg ctgccttaca 3361 ggcttcgact ccactacgtt caaccataga tacgacaaca cttgcaccca attgatctgc 3421 acgagatttg atagcggcaa caatttgaga aggggcatga agtgccaatg aagaggtagc 3481 aactccgata agcaaacttt gtttaccagc caactgttgg gcaacacgat ttgggatgta 3541 attaagttca gccattgcag cttcaacctt ttcacgagtt tttgctgata catgactagc 3601 ctgattgact acacgactga ctgtttggta agagactcct gcgtattctg cgacatcgta 3661 aagggtgact ggtttcatta attttcctcc ttatttattt agatccttgc atgtatagtt 3721 attataccag ggggacagtg caatgtcaag ctgaaatctt cgagcaggat ctagggtcgc 3781 tgaaacttct gatgacgtac agtcatattt cttctctttt tattatatca cagttttaaa 3841 tctagcttta ctagattcac cgctactatc tatttattcg gaaaaaagac gaaaaacctg 3901 agaatcatct caggcttggt cattaaattt ttttctcaat atcgaaaagt ggagaaagtg 3961 gtcgtttttc atgaatacgt acgatagcat cccctaggag atgagcgatt gaaatctgct 4021 caatcttatc aatcaaacgc tcttctggca gatagatggt atccaaaaca accaatttct 4081 taatagctga tttttggata ttgtccgtag caggaccaga aagaactggg tgcgtacagc 4141 ttgcatagac ttcaacagca ccagcttccg caagagcatc tgccgcatga caaatcgttc 4201 cagcggtatc aatcatatca tcaatcaaga tacaagtctt gccttcaacc ttaccgatga 4261 tgttcataac ttcactagag cgaaaaaacc ccgccgaagc ggggtttttt gcgactagtt 4321 ttctgcagta cacgatcgga gaccacccta tcaaaagaat cttgcttggc aaggttcttc 4381 tgctcgaaga tttcagcttg acattgcact gtccccctgg tataataact atacatgcaa 4441 ggatctaaat aaataaggag gaaaattaat ggctaaaatg agaatatcac cggaattgaa 4501 aaaactgatc gaaaaatacc gctgcgtaaa agatacggaa ggaatgtctc ctgctaaggt 4561 atataagctg gtgggagaaa atgaaaacct atatttaaaa atgacggaca gccggtataa 4621 agggaccacc tatgatgtgg aacgggaaaa ggacatgatg ctatggctgg aaggaaagct 4681 gcctgttcca aaggtcctgc actttgaacg gcatgatggc tggagcaatc tgctcatgag 4741 tgaggccgat ggcgtccttt gctcggaaga gtatgaagat gaacaaagcc ctgaaaagat 4801 tatcgagctg tatgcggagt gcatcaggct ctttcactcc atcgacatat cggattgtcc 4861 ctatacgaat agcttagaca gccgcttagc cgaattggat tacttactga ataacgatct 4921 ggccgatgtg gattgcgaaa actgggaaga agacactcca tttaaagatc cgcgcgagct 4981 gtatgatttt ttaaagacgg aaaagcccga agaggaactt gtcttttccc acggcgacct 5041 gggagacagc aacatctttg tgaaagatgg caaagtaagt ggctttattg atcttgggag 5101 aagcggcagg gcggacaagt ggtatgacat tgccttctgc gtccggtcga tcagggagga 5161 tatcggggaa gaacagtatg tcgagctatt ttttgactta ctggggatca agcctgattg 5221 ggagaaaata aaatattata ttttactgga tgaattgttt taggctaatt ttattgcaat 5281 aacaagtgct tacttttctg gacgggactc cgtccagttt tttgctagaa gctgtataat 5341 aaagttacaa agatgtacga ccacactgtt gtaaatcata gtgttcgcga atatattact 5401 gatagcattt ctacaaatac aattgaagag actggatagg ctttaaacga aatatgtcag 5461 tgttttggca ttcctagtct tcatatcatt taaagaattg tatagacaaa atttttttcc 5521 aatacagaca ctcgtaacaa ctgcttcatt tttctaccaa catatttagg aacaggataa 5581 gatagaagaa gattaatcca tagctcagtt ctataccaat ctaagacaaa taagctaaaa 5641 aaacgattga taataagcaa atagattcca aattttctct atctgctcat tttaataaac 5701 aatactagtg taactatcct tccagtcaga aacttgtcaa atcacaccga aaactcttct 5761 aaaatttatc tcgttaggca atcaagcaaa aactcgacga tagtacaaac attattatac 5821 agaattgact gcctaaatta tata //