LOCUS       sequence-170791-        6213 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Expression of sgRNA from a imyc promoter (Pimyc).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 6213)
  AUTHORS   Singh AK, Carette X, Potluri LP, Sharp JD, Xu R, Prisic S, Husson RN
  TITLE     Investigating essential gene function in Mycobacterium tuberculosis 
            using an efficient CRISPR interference system.
  JOURNAL   Nucleic Acids Res. 2016 Jul 12. pii: gkw625.
  PUBMED    27407107
REFERENCE   2  (bases 1 to 6213)
  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: "Nucleic 
            Acids Res. 2016 Jul 12. pii: gkw625."
COMMENT     Sequence Label: pRH2521
FEATURES             Location/Qualifiers
     source          1..6213
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     primer_bind     2603..2619
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     2604..2620
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     terminator      2763..2790
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     misc_RNA        3069..3144
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     terminator      complement(4154..4181)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     primer_bind     complement(4271..4288)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4442..5030)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4522..4541)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 aggatcaacg aggacagtcg cacgacgaag ttcttctgga tcgcgcccgt gctggaagca
       61 ctcaacctcg aagcgtgtgg ttgcggagcc atctagcaac cacacgaaac atgcgcaacg
      121 aaccgcgcaa cgaacaacgc ctagaactgg ccctagatga gctgactcgt atcgttggta
      181 aacctagttt gaccagcatg ttttaactac gttcggtgag ctgtcaacgg ggcctgtaac
      241 ggcacaacga accgtgcaac gagagtggcc acggatgcca ccacaggcac tacaacggag
      301 ttcgccacgt acatcaccac aaccaccgat tctggcggtg agctccccga tattcagcgg
      361 aaatggcttg gtatcgacca agattcgtag aaccccgtct cgtctggctg gtattcaaaa
      421 cgggcgcaac gaaacacgca acgagacagg catggcccaa accagaaaac tagcgtctac
      481 caggactttt acctgtccga cccgttgcaa cggaaccccc cacggaaccc ccgcgacacc
      541 cgctccccaa ttgcgttaga acagcggtgg attgtcggct tcgttgtggg ccttttgagc
      601 cgcttcctgt tctgccgcac gctctttcct cgcccgatag ccgagtcgct taacggtgtc
      661 cagatgcagc ccgaaatgtt tggccgtttg cggccaagag tggccctcgt cgtcgtgata
      721 ggcgcggatg cgttcgcggc gtgcagcctg ctcggcgagc cactcgctgc gttcctgcgc
      781 cacgagccgg acgacgtggc gttcggatag tccggtgatt cgagcgcctt cggcggcggt
      841 cacgcgccgc tttttgcgga cagtcggctg ccggttgtag ccgtcgctgt agccgtcgct
      901 catagcaatg cctccatggc tgacgcggac tttgcgcgcc gcgcaactgt gctcgccgcc
      961 gtgcgcgctg ctgcgccctt ccgcgagatg gccgactggc gcgcactgag tgtggcctcg
     1021 tagaccacga tcccgtccgc ccaaatgcgc gacttggttg tgatccaacg ccaaatgctg
     1081 ttggcgatgg cgcggacctc gctgtccggt agcggtccgg gacacacgtc gttgcacggg
     1141 aattcggcgt ttcgcgcgtg gcactcggca tagatcgcgc ggccgagtcc gtccacgttc
     1201 cgggtcggca ggtagatccg catgagggcg ggacgatagg cccacaacct gacggaatcg
     1261 aacagtgcgc aattccgccc tagcggcgtc ggagccgctt tgtacgtggt ctgctgacgc
     1321 cagcgcggcg gtggcatgtt cgcgccgagc tcggcctcga tgtggctgag tgtgtagaga
     1381 tctgagtgga gccattccgt ttcccaggcg atgtggccgg ggtttttggt catgaggcct
     1441 gagtaactgc ggtcgccgtc gacggcgcgc cgaaggcctt cggcgcacgc cgccatgtat
     1501 gcgagcggct tacgccgcgc gtattcggtg cgtggaacag gggcgttgag tgcccacact
     1561 gcgtgtgcgt ggccgttggc gcgattgccc acgatcgcgt tgggcagcgg atgggacccc
     1621 cgggcgctga gcgctcggag cgctgcgtct ggatggtcta cgtccacgac cagcaggttt
     1681 gccagcgctg ttgggttcgc ctcgatgtac cggcggccta gggccgacgc gcggctttgg
     1741 cggtagatcc cctcgagcag atcgtcgctt gccagcggcc agtacggcag ccagagctgc
     1801 tcaaattcgt cggcgacgtg gctcacgctt ggtagtagac cacgattaat caccggtgta
     1861 tggtccgaca cgagctccaa gtcagatatt tcgctgaggg gccaccccac aactgcacac
     1921 tcccccgctc tcccgtcgag ccctggtggt ggaacaccag cgacagccga gcacccccaa
     1981 ccacctgtac caaccaggag gaacacatgc gtcgtttcga ggacgtttcc gggccgctga
     2041 gagccgctgt ggcggccgta cacgccgcct tagacccgtt agaccccctg ccgcctgaat
     2101 gcgcgggtac gagccacaca gcgcccgaac ttacggagct ggtgggctca cctggcttta
     2161 tggcgtacga atcggctgtg tgcgacctgt tgggcgaggt gaggtacgcg ctactcacgc
     2221 tggcaagggc gacagagccg ccccaccgag cccgcacggc cgcgcgcggt gtcaacaacc
     2281 gggtgagtcg tgcacaccag caggtgttcg aggcttggct cgaagtgcag gacatcgtgg
     2341 cgaacgccgc ccgatgagcc gcgccttacg ctggctgcca gccgttcgcg ggctggttgg
     2401 tgcagcgcgt cgagcggtta gaggccctgc ggtgttccac caccgcaggg cctcgccctt
     2461 tttaaggctg aatttgcttg tctccgaatc caactggctt gtccaagggt gtatctacgc
     2521 ttaatccaaa gttcaaacga ggggattaca catgaccaac ttcgataacg ttctcggctc
     2581 gatgatcccc gggggccccc cctcgaggtc gacggtatcg ataatattgg atcgtcgcac
     2641 cgggttaagc ctggagtgcg gtggtgcctg gtcggcattt tcgcagtcga gggctctcgt
     2701 gtagcctggg cgagttgccg acgcaggcga ccctcctgcc acggatccgt gattagctaa
     2761 gcagaaggct atcctgacgg atggcctttt tgcgtttaat actgtttaaa ctctagaaat
     2821 attggatcgt cggcaccgtc acggccgtgg gaggcggcac gatccgcgac gtgatgatcg
     2881 gccgcatccc cacggtgctg cgcagtgagc tctacgccat cccggcgttg atctgtgcgt
     2941 tcgcacgcac aggcccggtg tgagaagggt ctctgacgag cgggagaact ccctatcagt
     3001 gatagagttt gtcctcccta tcagtgatag ataggctatg ggagtgtctt cgagacagag
     3061 gaagacatgt tttagagcta gaaatagcaa gttaaaataa ggctagtccg ttatcaactt
     3121 gaaaaagtgg caccgagtcg gtgctttttt atcgatttaa ttaatatgca tcaattgatt
     3181 taaatactag tgttaactat ttaattgggg accctagagg tccccttttt tattttaaaa
     3241 attttttcaa ggggacctct agggtcccca gctggctagc gagtcatgag gtcaccatgg
     3301 tcattaggag ccgctctcgc acttcagctg cttctccaag ccgcagatga tcagctccag
     3361 gccgaacaag aaggccggct cggcgccctg gtggtcgaac agctcgatgg cctggcgcag
     3421 caacggcggc atgctgtcgg tggtcggggt ctcgcgctcc tccttggcga cctggtgctc
     3481 ctggtcctcc aacacgcagc ccagggtgaa gtggcccacg gccgacaggg cgtacaacgc
     3541 gttctccagc gagaagccct gctggcacag gaaggccaac tggttctcca gggtctcgta
     3601 ctgcttctcg gtcggccggg tgcccaggtg caccttggcg ccgtcgcggt ggctcagcaa
     3661 ggcgcagcgg aagctcttgg cgttgttgcg caggaagtcc tgccagctct cgccctccag
     3721 cgggcagaag tgggtgtggt ggcggtccaa catctcgatg gccagggcgt cgagcagggc
     3781 ccgcttgttc ttgacgtgcc agtacagggt cggctgctcg acgcccagct tctgggccag
     3841 cttgcgggtg gtcaggccct cgatgccgac ctcgttcagc agctccaggg cggagttgat
     3901 gaccttggac ttgtccaggc gggacattgc gaagtgattc ctcctgatgg taccggatcc
     3961 gtggcaggag ggtcgcctgc gtcggcaact cgcccaggct acacgagagc cctcgactgc
     4021 gaaaatgccg accaggcacc accgcactcc aggcttaacc cggtgcgacg atccaatatt
     4081 acgagatctg atatcctcga ggcatgccac gtgcctgcag gtttaaacgc ggccgcaaac
     4141 agtattaaac gcaaaaaggc catccgtcag gatggccttc tgcttagcta atcaactagt
     4201 tctagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct
     4261 cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat
     4321 cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga
     4381 acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt
     4441 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt
     4501 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc
     4561 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa
     4621 gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct
     4681 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta
     4741 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg
     4801 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc
     4861 ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta
     4921 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg
     4981 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt
     5041 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg
     5101 tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta
     5161 aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg
     5221 aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgacg
     5281 tggccgacca gcccgtcatc gtcaacgcct gatccgcggt gcggacaggc cgtgtcgcga
     5341 ccggccgtgc ggaattaagc cggcccgtac cctgtgaata gaggtccgct gtgacacaag
     5401 aatccctgtt acttctcgac cgtattgatt cggatgattc ctacgcgagc ctgcggaacg
     5461 accaggaatt ctgggagccg ctggcccgcc gagccctgga ggagctcggg ctgccggtgc
     5521 cgccggtgct gcgggtgccc ggcgagagca ccaaccccgt actggtcggc gagcccggcc
     5581 cggtgatcaa gctgttcggc gagcactggt gcggtccgga gagcctcgcg tcggagtcgg
     5641 aggcgtacgc ggtcctggcg gacgccccgg tgccggtgcc ccgcctcctc ggccgcggcg
     5701 agctgcggcc cggcaccgga gcctggccgt ggccctacct ggtgatgagc cggatgaccg
     5761 gcaccacctg gcggtccgcg atggacggca cgaccgaccg gaacgcgctg ctcgccctgg
     5821 cccgcgaact cggccgggtg ctcggccggc tgcacagggt gccgctgacc gggaacaccg
     5881 tgctcaccac ccattccgag atcttcccgg aactgctgcg ggaacgccgc gcggcgaccg
     5941 tcgaggacca ccgcgggtgg ggctacctct cgccccggct gctggaccgc ctggaggact
     6001 ggctgccgga cgtggacacg ctgctggccg gccgcgaacc ccggttcgtc cacggcgacc
     6061 tgcacgggac caacatcttc gtggacctgg ccgcgaccga ggtcaccggg atcgtcgact
     6121 tcaccgacgt ctatgcggga gactcccgct acagcctggt gcaactgcat ctcaacgcct
     6181 tccggggcga ccgcgagatc ctggccgcgc tgc
//
