LOCUS       sequence-172946-        5844 bp    DNA     circular SYN 15-DEC-2025
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    .
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 Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Mol Syst 
            Biol."; date: "2017-05-10"; volume: "13(5):931. doi"; pages: " 
            10.15252/msb.20167449"
COMMENT     Sequence Label: pPEPY-PF6-lacI
FEATURES             Location/Qualifiers
     source          1..5844
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     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
//
