LOCUS       sequence-339553-        8631 bp    DNA     circular SYN 14-APR-2025
DEFINITION  Optimized M. smegmatis CRISPRi plasmid; L5 integrating..
ACCESSION   .
VERSION     .
KEYWORDS    .
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 Apr 14, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Cell. 2021 
            Jul 16. pii: S0092-8674(21)00824-2. doi: 
            10.1016/j.cell.2021.06.033."
COMMENT     Sequence Label: plRL117
FEATURES             Location/Qualifiers
     source          1..8631
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      606..633
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     primer_bind     complement(3399..3418)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             3410..4225
                     /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(3483..3502)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      complement(4938..5526)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5018..5037)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     protein_bind    5709..5727
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    5739..5757
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
ORIGIN
        1 caacaaggtg gtgctgcagt cggtgtcgcc gtggcgcgcc gacgtgtact tcaacaagac
       61 caccggcaag tacgagatcc tgggcctgaa gtacgccgac ctgcagttcg acaagggcac
      121 cggcacctac aagatctcgc aggagaagta caacgacatc aagaagaagg agggcgtgga
      181 ctcggactcg gagttcaagt tcaccctgta caagaacgac ctgctgctgg tgaaggacac
      241 cgagaccaag gagcagcagc tgttccgctt cctgtcgcgc accatgccga agcagaagca
      301 ctacgtggag ctgaagccgt acgacaagca gaagttcgag ggcggcgagg ccctgatcaa
      361 ggtgctgggc aacgtggcca actcgggcca gtgcaagaag ggcctgggca agtcgaacat
      421 ctcgatctac aaggtgcgca ccgacgtgct gggcaaccag cacatcatca agaacgaggg
      481 cgacaagccg aagctggact tctaaatact ctggccattt ttctttaccg cctttgagtg
      541 agctgatacc gctcgccgca gccgaacgac cgagcgcaac gcgtgcggcc gctgattagc
      601 taagcagaag gccatcctga cggatggcct ttttctagaa aaaaaaaaag cgccgcaact
      661 gcggcgcttt ttttttttta attaagtgat agattgcgat gcgtgtgctc ggttgatata
      721 atctgtcaga tcaggaggac ttcgcatgtc gcggttggat aagtccaagg tcatcaactc
      781 ggcgctggag ctgctgaacg aggtcggtat cgagggactg accacgcgca agttggccca
      841 aaagctcggc gtggaacaac cgaccctcta ctggcacgtg aagaacaagc gcgcgctgct
      901 ggatgcgttg gccatcgaga tgctggatcg ccatcacacg cacttttgcc ccctggaggg
      961 cgagagctgg caggattttc tgcggaacaa cgcgaagtcg ttccggtgcg ccctcctgag
     1021 ccatcgggat ggcgccaaag tccacttggg cacccgcccg accgagaagc aatacgagac
     1081 cctggagaac cagctggcct tcctctgtca acagggtttt tcgttggaga acgccctgta
     1141 tgccctgtcc gccgtgggac acttcacctt gggctgcgtg ttggaggatc aggaacatca
     1201 ggtggcgaaa gaggaacggg agacccccac gacggattcg atgcccccgt tgttgcgcca
     1261 ggcgatcgaa ctgtttgacc accagggcgc cgaacccgcc tttttgttcg ggctggagct
     1321 gatcatctgc gggctggaaa agcaactgaa gtgcgaaagc ggttcgtgat aacgccgaaa
     1381 accccgcttc ggcggggttt tgccgcggtc tgcgttgtcg ggaagattta aaataaaaaa
     1441 ggggacctct agggtcccca attaaatagt taacactagt attaaaaaaa aaagccccgc
     1501 gattgcgggg cttttttttt tcgacggcga gggttccgac cgctgcaact cccggtgcaa
     1561 ccttgtcccg gtctattctc ttcactgcac cagctccaat ctggtgtgaa tgcccctcgt
     1621 ctgttcgcgc aggcgggggg ctctattcgt ttgtcagcat cgaaagtagc cagatcaggg
     1681 atgcgttgca accgcgtatg cccaggtcag aagagtcgca caagagttgc agacccctgg
     1741 aaagaaaaat ggccagaggg cgaaaacacc ctctgaccag cggagcgggc gacgggaatc
     1801 gaacccgcgt agctagtttg gaagaatggg tgtctgccga ccacatatat gggccggtca
     1861 agataggttt ttaccccctc tcggctgcat cctctaagtg gaaagaaatt gcaggtcgta
     1921 gaagcgcgtt gaagcctgag agttgcacag gagttgcaac ccggtagcct tgttcacgac
     1981 gagaggagac ctagttggca cgtcgcggat ggggatcgct gaagactcag cgcagcggga
     2041 ggatccaagc ctcatacgtc aacccgcagg acggtgtgag gtactacgcg ctgcagacct
     2101 acgacaacaa gatggacgcc gaagcctggc tcgcgggcga gaagcggctc atcgagatgg
     2161 agacctggac ccctccacag gaccgggcga agaaggcagc cgccagcgcc atcacgctgg
     2221 aggagtacac ccggaagtgg ctcgtggagc gcgacctcgc agacggcacc agggatctgt
     2281 acagcgggca cgcggagcgc cgcatctacc cggtgctagg tgaagtggcg gtcacagaga
     2341 tgacgccagc tctggtgcgt gcgtggtggg ccgggatggg taggaagcac ccgactgccc
     2401 gccggcatgc ctacaacgtc ctccgggcgg tgatgaacac agcggtcgag gacaagctga
     2461 tcgcagagaa cccgtgccgg atcgagcaga aggcagccga tgagcgcgac gtagaggcgc
     2521 tgacgcctga ggagctggac atcgtcgccg ctgagatctt cgtgcactac cggatcgcgg
     2581 catacatcct ggcgtggacg agcctccggt tcggagagct gatcgagctt cgccgcaagg
     2641 acatcgtgga cgacggcatg acgatgaagc tccgggtgcg ccgtggcgct tcccgcgtgg
     2701 ggaacaagat cgtcgttggc aacgccaaga ccgtccggtc gaagcgtcct gtgacggttc
     2761 cgcctcacgt cgcggagatg atccgagcgc acatgaagga ccgtacgaag atgaacaagg
     2821 gccccgaggc attcctggtg accacgacgc agggcaaccg gctgtcgaag tccgcgttca
     2881 ccaagtcgct gaagcgtggc tacgccaaga tcggtcggcc ggaactccgc atccacgacc
     2941 tccgcgctgt cggcgctacg ttcgccgctc aggcaggtgc gacgaccaag gagctgatgg
     3001 cccgcctcgg tcacacgact cctaggatgg cgatgaagta ccagatggcg tctgaggccc
     3061 gcgacgaggc tatcgctgag gcgatgtcca agctggccaa gacctcctga aacgcaaaaa
     3121 gcccccctcc caaggacact gagtcctaaa gaggggggtt tcttgtcagt acgcgaagaa
     3181 ccacgcctgg ccgcgagcgc cagcaccgcc gctctgtgcg gagacctggg caccagcccc
     3241 gccgccgcca ggagcattgc cgttcccgcc agctagccac ctgacgtcgg ggggggggga
     3301 aagccacgtt gtgtctcaaa atctctgatg ttacattgca caagataaaa atatatcatc
     3361 atgaacaata aaactgtctg cttacataaa cagtaataca aggggtgtta tgagccatat
     3421 tcaacgggaa acgtcttgct cgaggccgcg attaaattcc aacatggatg ctgatttata
     3481 tgggtataaa tgggctcgcg ataatgtcgg gcaatcaggt gcgacaatct atcgcttgta
     3541 tgggaagccc catgcgccag agttgtttct gaaacatggc aaaggtagcg ttgccaatga
     3601 tgttacagat gagatggtca gactaaactg gctgacggaa tttatgcctc ttccgaccat
     3661 caagcatttt atccgtactc ctgatgatgc atggttactc accactgcga tccccgggaa
     3721 aacagcattc caggtattag aagaatatcc tgattcaggt gaaaatattg ttgatgcgct
     3781 ggcagtgttc ctgcgccggt tgcattcgat tcctgtttgt aattgtcctt ttaacagcga
     3841 tcgcgtattt cgcctcgctc aggcgcaatc acgaatgaat aacggtttgg ttgatgcgag
     3901 tgattttgat gacgagcgta atggctggcc tgttgaacaa gtctggaaag aaatgcataa
     3961 tcttttgcca ttctcaccgg attcagtcgt cactcatggt gatttctcac ttgataacct
     4021 tatttttgac gaggggaaat taataggttg tattgatgtt ggacgagtcg gaatcgcaga
     4081 ccgataccag gatcttgcca tcctatggaa ctgcctcggt gagttttctc cttcattaca
     4141 gaaacggctt tttcaaaaat atggtattga taatcctgat atgaataaat tgcagtttca
     4201 tttgatgctc gatgagtttt tctaatcaga attggttaat tggttgtaac actggcagag
     4261 cattacgctg acttgacggg acggcggctt tgttgaataa atcgaacttt tgctgagttg
     4321 aaggatcaga tcacgcatct tcccgacaac gcagaccgtt ccgtggcaaa gcaaaagttc
     4381 aaaatcacca actggtccac ctacaacaaa gctctcacca accgtggctc cctcacgata
     4441 tcaataaacg aaaggctcag tcgaaagact gggcctttcg tttatctgtt gtttgaaaaa
     4501 aaaaagcgcc gcaactgcgg cgcttttttt tttgaattct ctgaccaggg aaaatagccc
     4561 tctgacctgg ggatttgcga tccctatcag tgatagatat aatctgggag gagacgatta
     4621 atgcgtctcg gtttttgtac tcgaaagaag ctacaaagat aaggcttcat gccgaaatca
     4681 acaccctgtc attttatggc agggtgtttt ttttttgtcg acttggggac cctagaggtc
     4741 ccctttttta tttttttgga tgaagagcat tatcaatggc tcttcacagg aagcttaaaa
     4801 aaaaaagccc cgcgattgcg gggctttttt tttttaatac ggttatccac agaatcaggg
     4861 gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag
     4921 gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga
     4981 cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct
     5041 ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc
     5101 tttctcccct cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg
     5161 gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc
     5221 tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca
     5281 ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag
     5341 ttcttgaagt ggtggcctaa ctacggctac actagaagga cagtatttgg tatctgcgct
     5401 ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc
     5461 accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga
     5521 tctcaagaag atcctttgat cttttctacg gggtctgacg ctcatggaac aataaacgaa
     5581 aggctcagtc gaaagactgg gcctttcgtt tatctgttgt ttgaaaaaaa aaagccccgc
     5641 gattgcgggg cttttttttt tcatatgtct gaccagggaa aatagccctc tgacctgggg
     5701 atttgacttc cctatcagtg atagagataa tctgggagtc cctatcagtg atagagaagg
     5761 cgctatcgat atgtcggacc tggtgctggg cctggccatc ggcatcggct cggtgggcgt
     5821 gggcatcctg aacaaggtga ccggcgagat catccacaag aactcgcgca tcttcccggc
     5881 cgcccaggcc gagaacaacc tggtgcgccg caccaaccgc cagggccgcc gcctggcccg
     5941 ccgcaagaag caccgccgcg tgcgcctgaa ccgcctgttc gaggagtcgg gcctgatcac
     6001 cgacttcacc aagatctcga tcaacctgaa cccgtaccag ctgcgcgtga agggcctgac
     6061 cgacgagctg tcgaacgagg agctgttcat cgccctgaag aacatggtga agcaccgcgg
     6121 catctcgtac ctggacgacg cctcggacga cggcaactcg tcggtgggcg actacgccca
     6181 gatcgtgaag gagaactcga agcagctgga gaccaagacc ccgggccaga tccagctgga
     6241 gcgctaccag acctacggcc agctgcgcgg cgacttcacc gtggagaagg acggcaagaa
     6301 gcaccgcctg atcaacgtgt tcccgacctc ggcctaccgc tcggaggccc tgcgcatcct
     6361 gcagacccag caggagttca acccgcagat caccgacgag ttcatcaacc gctacctgga
     6421 gatcctgacc ggcaagcgca agtactacca cggcccgggc aacgagaagt cgcgcaccga
     6481 ctacggccgc taccgcacct cgggcgagac cctggacaac atcttcggca tcctgatcgg
     6541 caagtgcacc ttctacccgg acgagttccg cgccgccaag gcctcgtaca ccgcccagga
     6601 gttcaacctg ctgaacgacc tgaacaacct gaccgtgccg accgagacca agaagctgtc
     6661 gaaggagcag aagaaccaga tcatcaacta cgtgaagaac gagaaggcca tgggcccggc
     6721 caagctgttc aagtacatcg ccaagctgct gtcgtgcgac gtggccgaca tcaagggcta
     6781 ccgcatcgac aagtcgggca aggccgagat ccacaccttc gaggcctacc gcaagatgaa
     6841 gaccctggag accctggaca tcgagcagat ggaccgcgag accctggaca agctggccta
     6901 cgtgctgacc ctgaacaccg agcgcgaggg catccaggag gccctggagc acgagttcgc
     6961 cgacggctcg ttctcgcaga agcaggtgga cgagctggtg cagttccgca aggccaactc
     7021 gtcgatcttc ggcaagggct ggcacaactt ctcggtgaag ctgatgatgg agctgatccc
     7081 ggagctgtac gagacctcgg aggagcagat gaccatcctg acccgcctgg gcaagcagaa
     7141 gaccacctcg tcgtcgaaca agaccaagta catcgacgag aagctgctga ccgaggagat
     7201 ctacaacccg gtggtggcca agtcggtgcg ccaggccatc aagatcgtga acgccgccat
     7261 caaggagtac ggcgacttcg acaacatcgt gatcgagatg gcccgcgaga ccaacgagga
     7321 cgacgagaag aaggccatcc agaagatcca gaaggccaac aaggacgaga aggacgccgc
     7381 catgctgaag gccgccaacc agtacaacgg caaggccgag ctgccgcact cggtgttcca
     7441 cggccacaag cagctggcca ccaagatccg cctgtggcac cagcagggcg agcgctgcct
     7501 gtacaccggc aagaccatct cgatccacga cctgatcaac aactcgaacc agttcgaggt
     7561 ggacgccatc ctgccgctgt cgatcacctt cgacgactcg ctggccaaca aggtgctggt
     7621 gtacgccacc gccaaccagg agaagggcca gcgcaccccg taccaggccc tggactcgat
     7681 ggacgacgcc tggtcgttcc gcgagctgaa ggccttcgtg cgcgagtcga agaccctgtc
     7741 gaacaagaag aaggagtacc tgctgaccga ggaggacatc tcgaagttcg acgtgcgcaa
     7801 gaagttcatc gagcgcaacc tggtggacac ccgctacgcc tcgcgcgtgg tgctgaacgc
     7861 cctgcaggag cacttccgcg cccacaagat cgacaccaag gtgtcggtgg tgcgcggcca
     7921 gttcacctcg cagctgcgcc gccactgggg catcgagaag acccgcgaca cctaccacca
     7981 ccacgccgtg gacgccctga tcatcgccgc ctcgtcgcag ctgaacctgt ggaagaagca
     8041 gaagaacacc ctggtgtcgt actcggagga ccagctgctg gacatcgaga ccggcgagct
     8101 gatctcggac gacgagtaca aggagtcggt gttcaaggcc ccgtaccagc acttcgtgga
     8161 caccctgaag tcgaaggagt tcgaggactc gatcctgttc tcgtaccagg tggactcgaa
     8221 gttcaaccgc aagatctcgg acgccaccat ctacgccacc cgccaggcca aggtgggcaa
     8281 ggacaaggcc gacgagacct acgtgctggg caagatcaag gacatctaca cccaggacgg
     8341 ctacgacgcc ttcatgaaga tctacaagaa ggacaagtcg aagttcctga tgtaccgcca
     8401 cgacccgcag accttcgaga aggtgatcga gccgatcctg gagaactacc cgaacaagca
     8461 gatcaacgac aagggcaagg aggtgccgtg caacccgttc ctgaagtaca aggaggagca
     8521 cggctacatc cgcaagtact cgaagaaggg caacggcccg gagatcaagt cgctgaagta
     8581 ctacgactcg aagctgggca accacatcga catcaccccg aaggactcga a
//
