plRL2

LOCUS       plRL2                8631 bp ds-DNA     circular SYN 10-AUG-2021
DEFINITION  Optimized M. tuberculosis CRISPRi plasmid; L5 integrating..
ACCESSION   .
VERSION     .
KEYWORDS    plRL2
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8631)
  AUTHORS   Bosch B, DeJesus MA, Poulton NC, Zhang W, Engelhart CA, Zaveri A, 
            Lavalette S, Ruecker N, Trujillo C, Wallach JB, Li S, Ehrt S, Chait 
            BT, Schnappinger D, Rock JM
  TITLE     Genome-wide gene expression tuning reveals diverse vulnerabilities 
            of M. tuberculosis.
  JOURNAL   Cell. 2021 Jul 16. pii: S0092-8674(21)00824-2. doi: 
            10.1016/j.cell.2021.06.033.
  PUBMED    34297925
REFERENCE   2  (bases 1 to 8631)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Aug 10, 2021 from SnapGene Server 1.1.58
            http://www.biofeng.com
FEATURES             Location/Qualifiers
     source          1..8631
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     rep_origin      complement(496..1084)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(576..595)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     protein_bind    1267..1285
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1297..1315
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     terminator      4795..4822
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     primer_bind     complement(7588..7607)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             7599..8414
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPHAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHNLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     complement(7672..7691)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
ORIGIN
        1 aataaacgaa aggctcagtc gaaagactgg gcctttcgtt tatctgttgt ttgaaaaaaa
       61 aaagcgccgc aactgcggcg cttttttttt tgaattctct gaccagggaa aatagccctc
      121 tgacctgggg atttgcgatc cctatcagtg atagatataa tctgggagga gacgattaat
      181 gcgtctcggt ttttgtactc gaaagaagct acaaagataa ggcttcatgc cgaaatcaac
      241 accctgtcat tttatggcag ggtgtttttt ttttgtcgac ttggggaccc tagaggtccc
      301 cttttttatt tttttggatg aagagcatta tcaatggctc ttcacaggaa gcttaaaaaa
      361 aaaagccccg cgattgcggg gctttttttt tttaatacgg ttatccacag aatcagggga
      421 taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc
      481 cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg
      541 ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg
      601 aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt
      661 tctcccctcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt
      721 gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg
      781 cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact
      841 ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt
      901 cttgaagtgg tggcctaact acggctacac tagaaggaca gtatttggta tctgcgctct
      961 gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac
     1021 cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc
     1081 tcaagaagat cctttgatct tttctacggg gtctgacgct catggaacaa taaacgaaag
     1141 gctcagtcga aagactgggc ctttcgttta tctgttgttt gaaaaaaaaa agccccgcga
     1201 ttgcggggct tttttttttc atatgtctga ccagggaaaa tagccctctg acctggggat
     1261 ttgacttccc tatcagtgat agagataatc tgggagtccc tatcagtgat agagaacgcg
     1321 gtatcgatat gtcggacctg gtgctgggcc tggccatcgg catcggctcg gtgggcgtgg
     1381 gcatcctgaa caaggtgacc ggcgagatca tccacaagaa ctcgcgcatc ttcccggccg
     1441 cccaggccga gaacaacctg gtgcgccgca ccaaccgcca gggccgccgc ctggcccgcc
     1501 gcaagaagca ccgccgcgtg cgcctgaacc gcctgttcga ggagtcgggc ctgatcaccg
     1561 acttcaccaa gatctcgatc aacctgaacc cgtaccagct gcgcgtgaag ggcctgaccg
     1621 acgagctgtc gaacgaggag ctgttcatcg ccctgaagaa catggtgaag caccgcggca
     1681 tctcgtacct ggacgacgcc tcggacgacg gcaactcgtc ggtgggcgac tacgcccaga
     1741 tcgtgaagga gaactcgaag cagctggaga ccaagacccc gggccagatc cagctggagc
     1801 gctaccagac ctacggccag ctgcgcggcg acttcaccgt ggagaaggac ggcaagaagc
     1861 accgcctgat caacgtgttc ccgacctcgg cctaccgctc ggaggccctg cgcatcctgc
     1921 agacccagca ggagttcaac ccgcagatca ccgacgagtt catcaaccgc tacctggaga
     1981 tcctgaccgg caagcgcaag tactaccacg gcccgggcaa cgagaagtcg cgcaccgact
     2041 acggccgcta ccgcacctcg ggcgagaccc tggacaacat cttcggcatc ctgatcggca
     2101 agtgcacctt ctacccggac gagttccgcg ccgccaaggc ctcgtacacc gcccaggagt
     2161 tcaacctgct gaacgacctg aacaacctga ccgtgccgac cgagaccaag aagctgtcga
     2221 aggagcagaa gaaccagatc atcaactacg tgaagaacga gaaggccatg ggcccggcca
     2281 agctgttcaa gtacatcgcc aagctgctgt cgtgcgacgt ggccgacatc aagggctacc
     2341 gcatcgacaa gtcgggcaag gccgagatcc acaccttcga ggcctaccgc aagatgaaga
     2401 ccctggagac cctggacatc gagcagatgg accgcgagac cctggacaag ctggcctacg
     2461 tgctgaccct gaacaccgag cgcgagggca tccaggaggc cctggagcac gagttcgccg
     2521 acggctcgtt ctcgcagaag caggtggacg agctggtgca gttccgcaag gccaactcgt
     2581 cgatcttcgg caagggctgg cacaacttct cggtgaagct gatgatggag ctgatcccgg
     2641 agctgtacga gacctcggag gagcagatga ccatcctgac ccgcctgggc aagcagaaga
     2701 ccacctcgtc gtcgaacaag accaagtaca tcgacgagaa gctgctgacc gaggagatct
     2761 acaacccggt ggtggccaag tcggtgcgcc aggccatcaa gatcgtgaac gccgccatca
     2821 aggagtacgg cgacttcgac aacatcgtga tcgagatggc ccgcgagacc aacgaggacg
     2881 acgagaagaa ggccatccag aagatccaga aggccaacaa ggacgagaag gacgccgcca
     2941 tgctgaaggc cgccaaccag tacaacggca aggccgagct gccgcactcg gtgttccacg
     3001 gccacaagca gctggccacc aagatccgcc tgtggcacca gcagggcgag cgctgcctgt
     3061 acaccggcaa gaccatctcg atccacgacc tgatcaacaa ctcgaaccag ttcgaggtgg
     3121 acgccatcct gccgctgtcg atcaccttcg acgactcgct ggccaacaag gtgctggtgt
     3181 acgccaccgc caaccaggag aagggccagc gcaccccgta ccaggccctg gactcgatgg
     3241 acgacgcctg gtcgttccgc gagctgaagg ccttcgtgcg cgagtcgaag accctgtcga
     3301 acaagaagaa ggagtacctg ctgaccgagg aggacatctc gaagttcgac gtgcgcaaga
     3361 agttcatcga gcgcaacctg gtggacaccc gctacgcctc gcgcgtggtg ctgaacgccc
     3421 tgcaggagca cttccgcgcc cacaagatcg acaccaaggt gtcggtggtg cgcggccagt
     3481 tcacctcgca gctgcgccgc cactggggca tcgagaagac ccgcgacacc taccaccacc
     3541 acgccgtgga cgccctgatc atcgccgcct cgtcgcagct gaacctgtgg aagaagcaga
     3601 agaacaccct ggtgtcgtac tcggaggacc agctgctgga catcgagacc ggcgagctga
     3661 tctcggacga cgagtacaag gagtcggtgt tcaaggcccc gtaccagcac ttcgtggaca
     3721 ccctgaagtc gaaggagttc gaggactcga tcctgttctc gtaccaggtg gactcgaagt
     3781 tcaaccgcaa gatctcggac gccaccatct acgccacccg ccaggccaag gtgggcaagg
     3841 acaaggccga cgagacctac gtgctgggca agatcaagga catctacacc caggacggct
     3901 acgacgcctt catgaagatc tacaagaagg acaagtcgaa gttcctgatg taccgccacg
     3961 acccgcagac cttcgagaag gtgatcgagc cgatcctgga gaactacccg aacaagcaga
     4021 tcaacgacaa gggcaaggag gtgccgtgca acccgttcct gaagtacaag gaggagcacg
     4081 gctacatccg caagtactcg aagaagggca acggcccgga gatcaagtcg ctgaagtact
     4141 acgactcgaa gctgggcaac cacatcgaca tcaccccgaa ggactcgaac aacaaggtgg
     4201 tgctgcagtc ggtgtcgccg tggcgcgccg acgtgtactt caacaagacc accggcaagt
     4261 acgagatcct gggcctgaag tacgccgacc tgcagttcga caagggcacc ggcacctaca
     4321 agatctcgca ggagaagtac aacgacatca agaagaagga gggcgtggac tcggactcgg
     4381 agttcaagtt caccctgtac aagaacgacc tgctgctggt gaaggacacc gagaccaagg
     4441 agcagcagct gttccgcttc ctgtcgcgca ccatgccgaa gcagaagcac tacgtggagc
     4501 tgaagccgta cgacaagcag aagttcgagg gcggcgaggc cctgatcaag gtgctgggca
     4561 acgtggccaa ctcgggccag tgcaagaagg gcctgggcaa gtcgaacatc tcgatctaca
     4621 aggtgcgcac cgacgtgctg ggcaaccagc acatcatcaa gaacgagggc gacaagccga
     4681 agctggactt ctaaatactc tggccatttt tctttaccgc ctttgagtga gctgataccg
     4741 ctcgccgcag ccgaacgacc gagcgcaacg cgtgcggccg ctgattagct aagcagaagg
     4801 ccatcctgac ggatggcctt tttctagaaa aaaaaaaagc gccgcaactg cggcgctttt
     4861 ttttttttaa ttaagtgata gattgcgatg cgtgtgctcg gttgatataa tctgtcagat
     4921 caggaggact tcgcatgtcg cggttggata agtccaaggt catcaactcg gcgctggagc
     4981 tgctgaacga ggtcggtatc gagggactga ccacgcgcaa gttggcccaa aagctcggcg
     5041 tggaacaacc gaccctctac tggcacgtga agaacaagcg cgcgctgctg gatgcgttgg
     5101 ccatcgagat gctggatcgc catcacacgc acttttgccc cctggagggc gagagctggc
     5161 aggattttct gcggaacaac gcgaagtcgt tccggtgcgc cctcctgagc catcgggatg
     5221 gcgccaaagt ccacttgggc acccgcccga ccgagaagca atacgagacc ctggagaacc
     5281 agctggcctt cctctgtcaa cagggttttt cgttggagaa cgccctgtat gccctgtccg
     5341 ccgtgggaca cttcaccttg ggctgcgtgt tggaggatca ggaacatcag gtggcgaaag
     5401 aggaacggga gacccccacg acggattcga tgcccccgtt gttgcgccag gcgatcgaac
     5461 tgtttgacca ccagggcgcc gaacccgcct ttttgttcgg gctggagctg atcatctgcg
     5521 ggctggaaaa gcaactgaag tgcgaaagcg gttcgtgata acgccgaaaa ccccgcttcg
     5581 gcggggtttt gccgcggtct gcgttgtcgg gaagatttaa aataaaaaag gggacctcta
     5641 gggtccccaa ttaaatagtt aacactagta ttaaaaaaaa aagccccgcg attgcggggc
     5701 tttttttttt cgacggcgag ggttccgacc gctgcaactc ccggtgcaac cttgtcccgg
     5761 tctattctct tcactgcacc agctccaatc tggtgtgaat gcccctcgtc tgttcgcgca
     5821 ggcggggggc tctattcgtt tgtcagcatc gaaagtagcc agatcaggga tgcgttgcaa
     5881 ccgcgtatgc ccaggtcaga agagtcgcac aagagttgca gacccctgga aagaaaaatg
     5941 gccagagggc gaaaacaccc tctgaccagc ggagcgggcg acgggaatcg aacccgcgta
     6001 gctagtttgg aagaatgggt gtctgccgac cacatatatg ggccggtcaa gataggtttt
     6061 taccccctct cggctgcatc ctctaagtgg aaagaaattg caggtcgtag aagcgcgttg
     6121 aagcctgaga gttgcacagg agttgcaacc cggtagcctt gttcacgacg agaggagacc
     6181 tagttggcac gtcgcggatg gggatcgctg aagactcagc gcagcgggag gatccaagcc
     6241 tcatacgtca acccgcagga cggtgtgagg tactacgcgc tgcagaccta cgacaacaag
     6301 atggacgccg aagcctggct cgcgggcgag aagcggctca tcgagatgga gacctggacc
     6361 cctccacagg accgggcgaa gaaggcagcc gccagcgcca tcacgctgga ggagtacacc
     6421 cggaagtggc tcgtggagcg cgacctcgca gacggcacca gggatctgta cagcgggcac
     6481 gcggagcgcc gcatctaccc ggtgctaggt gaagtggcgg tcacagagat gacgccagct
     6541 ctggtgcgtg cgtggtgggc cgggatgggt aggaagcacc cgactgcccg ccggcatgcc
     6601 tacaacgtcc tccgggcggt gatgaacaca gcggtcgagg acaagctgat cgcagagaac
     6661 ccgtgccgga tcgagcagaa ggcagccgat gagcgcgacg tagaggcgct gacgcctgag
     6721 gagctggaca tcgtcgccgc tgagatcttc gtgcactacc ggatcgcggc atacatcctg
     6781 gcgtggacga gcctccggtt cggagagctg atcgagcttc gccgcaagga catcgtggac
     6841 gacggcatga cgatgaagct ccgggtgcgc cgtggcgctt cccgcgtggg gaacaagatc
     6901 gtcgttggca acgccaagac cgtccggtcg aagcgtcctg tgacggttcc gcctcacgtc
     6961 gcggagatga tccgagcgca catgaaggac cgtacgaaga tgaacaaggg ccccgaggca
     7021 ttcctggtga ccacgacgca gggcaaccgg ctgtcgaagt ccgcgttcac caagtcgctg
     7081 aagcgtggct acgccaagat cggtcggccg gaactccgca tccacgacct ccgcgctgtc
     7141 ggcgctacgt tcgccgctca ggcaggtgcg acgaccaagg agctgatggc ccgcctcggt
     7201 cacacgactc ctaggatggc gatgaagtac cagatggcgt ctgaggcccg cgacgaggct
     7261 atcgctgagg cgatgtccaa gctggccaag acctcctgaa acgcaaaaag cccccctccc
     7321 aaggacactg agtcctaaag aggggggttt cttgtcagta cgcgaagaac cacgcctggc
     7381 cgcgagcgcc agcaccgccg ctctgtgcgg agacctgggc accagccccg ccgccgccag
     7441 gagcattgcc gttcccgcca gctagccacc tgacgtcggg ggggggggaa agccacgttg
     7501 tgtctcaaaa tctctgatgt tacattgcac aagataaaaa tatatcatca tgaacaataa
     7561 aactgtctgc ttacataaac agtaatacaa ggggtgttat gagccatatt caacgggaaa
     7621 cgtcttgctc gaggccgcga ttaaattcca acatggatgc tgatttatat gggtataaat
     7681 gggctcgcga taatgtcggg caatcaggtg cgacaatcta tcgcttgtat gggaagcccc
     7741 atgcgccaga gttgtttctg aaacatggca aaggtagcgt tgccaatgat gttacagatg
     7801 agatggtcag actaaactgg ctgacggaat ttatgcctct tccgaccatc aagcatttta
     7861 tccgtactcc tgatgatgca tggttactca ccactgcgat ccccgggaaa acagcattcc
     7921 aggtattaga agaatatcct gattcaggtg aaaatattgt tgatgcgctg gcagtgttcc
     7981 tgcgccggtt gcattcgatt cctgtttgta attgtccttt taacagcgat cgcgtatttc
     8041 gcctcgctca ggcgcaatca cgaatgaata acggtttggt tgatgcgagt gattttgatg
     8101 acgagcgtaa tggctggcct gttgaacaag tctggaaaga aatgcataat cttttgccat
     8161 tctcaccgga ttcagtcgtc actcatggtg atttctcact tgataacctt atttttgacg
     8221 aggggaaatt aataggttgt attgatgttg gacgagtcgg aatcgcagac cgataccagg
     8281 atcttgccat cctatggaac tgcctcggtg agttttctcc ttcattacag aaacggcttt
     8341 ttcaaaaata tggtattgat aatcctgata tgaataaatt gcagtttcat ttgatgctcg
     8401 atgagttttt ctaatcagaa ttggttaatt ggttgtaaca ctggcagagc attacgctga
     8461 cttgacggga cggcggcttt gttgaataaa tcgaactttt gctgagttga aggatcagat
     8521 cacgcatctt cccgacaacg cagaccgttc cgtggcaaag caaaagttca aaatcaccaa
     8581 ctggtccacc tacaacaaag ctctcaccaa ccgtggctcc ctcacgatat c
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