LOCUS       sequence-170793-        6624 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of sgRNA from a imyc promoter (Pimyc).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6624)
  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 6624)
  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..6624
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     37..53
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     38..54
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     terminator      197..224
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     misc_RNA        503..578
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     terminator      complement(1588..1615)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     primer_bind     complement(1705..1722)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1876..2464)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1956..1975)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 aacgttctcg gctcgatgat ccccgggggc cccccctcga ggtcgacggt atcgataata
       61 ttggatcgtc gcaccgggtt aagcctggag tgcggtggtg cctggtcggc attttcgcag
      121 tcgagggctc tcgtgtagcc tgggcgagtt gccgacgcag gcgaccctcc tgccacggat
      181 ccgtgattag ctaagcagaa ggctatcctg acggatggcc tttttgcgtt taatactgtt
      241 taaactctag aaatattgga tcgtcggcac cgtcacggcc gtgggaggcg gcacgatccg
      301 cgacgtgatg atcggccgca tccccacggt gctgcgcagt gagctctacg ccatcccggc
      361 gttgatctgt gcgttcgcac gcacaggccc ggtgtgagaa gggtctctga cgagcgggag
      421 aactccctat cagtgataga gtttgtcctc cctatcagtg atagataggc tatgggagtg
      481 tcttcgagac agaggaagac atgttttaga gctagaaata gcaagttaaa ataaggctag
      541 tccgttatca acttgaaaaa gtggcaccga gtcggtgctt ttttatcgat ttaattaata
      601 tgcatcaatt gatttaaata ctagtgttaa ctatttaatt ggggacccta gaggtcccct
      661 tttttatttt aaaaattttt tcaaggggac ctctagggtc cccagctggc tagcgagtca
      721 tgaggtcacc atggtcatta ggagccgctc tcgcacttca gctgcttctc caagccgcag
      781 atgatcagct ccaggccgaa caagaaggcc ggctcggcgc cctggtggtc gaacagctcg
      841 atggcctggc gcagcaacgg cggcatgctg tcggtggtcg gggtctcgcg ctcctccttg
      901 gcgacctggt gctcctggtc ctccaacacg cagcccaggg tgaagtggcc cacggccgac
      961 agggcgtaca acgcgttctc cagcgagaag ccctgctggc acaggaaggc caactggttc
     1021 tccagggtct cgtactgctt ctcggtcggc cgggtgccca ggtgcacctt ggcgccgtcg
     1081 cggtggctca gcaaggcgca gcggaagctc ttggcgttgt tgcgcaggaa gtcctgccag
     1141 ctctcgccct ccagcgggca gaagtgggtg tggtggcggt ccaacatctc gatggccagg
     1201 gcgtcgagca gggcccgctt gttcttgacg tgccagtaca gggtcggctg ctcgacgccc
     1261 agcttctggg ccagcttgcg ggtggtcagg ccctcgatgc cgacctcgtt cagcagctcc
     1321 agggcggagt tgatgacctt ggacttgtcc aggcgggaca ttgcgaagtg attcctcctg
     1381 atggtaccgg atccgtggca ggagggtcgc ctgcgtcggc aactcgccca ggctacacga
     1441 gagccctcga ctgcgaaaat gccgaccagg caccaccgca ctccaggctt aacccggtgc
     1501 gacgatccaa tattacgaga tctgatatcc tcgaggcatg ccacgtgcct gcaggtttaa
     1561 acgcggccgc aaacagtatt aaacgcaaaa aggccatccg tcaggatggc cttctgctta
     1621 gctaatcaac tagttctagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt
     1681 gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct
     1741 gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga
     1801 taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc
     1861 cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg
     1921 ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg
     1981 aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt
     2041 tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt
     2101 gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg
     2161 cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact
     2221 ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt
     2281 cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct
     2341 gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac
     2401 cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc
     2461 tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg
     2521 ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta
     2581 aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca
     2641 atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc
     2701 ctgactcccc gacgtggccg accagcccgt catcgtcaac gcctgatccg cggtgcggac
     2761 aggccgtgtc gcgaccggcc gtgcggaatt aagccggccc gtaccctgtg aatagaggtc
     2821 cgctgtgaca caagaatccc tgttacttct cgaccgtatt gattcggatg attcctacgc
     2881 gagcctgcgg aacgaccagg aattctggga gccgctggcc cgccgagccc tggaggagct
     2941 cgggctgccg gtgccgccgg tgctgcgggt gcccggcgag agcaccaacc ccgtactggt
     3001 cggcgagccc ggcccggtga tcaagctgtt cggcgagcac tggtgcggtc cggagagcct
     3061 cgcgtcggag tcggaggcgt acgcggtcct ggcggacgcc ccggtgccgg tgccccgcct
     3121 cctcggccgc ggcgagctgc ggcccggcac cggagcctgg ccgtggccct acctggtgat
     3181 gagccggatg accggcacca cctggcggtc cgcgatggac ggcacgaccg accggaacgc
     3241 gctgctcgcc ctggcccgcg aactcggccg ggtgctcggc cggctgcaca gggtgccgct
     3301 gaccgggaac accgtgctca ccacccattc cgagatcttc ccggaactgc tgcgggaacg
     3361 ccgcgcggcg accgtcgagg accaccgcgg gtggggctac ctctcgcccc ggctgctgga
     3421 ccgcctggag gactggctgc cggacgtgga cacgctgctg gccggccgcg aaccccggtt
     3481 cgtccacggc gacctgcacg ggaccaacat cttcgtggac ctggccgcga ccgaggtcac
     3541 cgggatcgtc gacttcaccg acgtctatgc gggagactcc cgctacagcc tggtgcaact
     3601 gcatctcaac gccttccggg gcgaccgcga gatcctggcc gcgctgctcg acggggcgca
     3661 gtggaagcgg accgaggact tcgcccgcga actgctcgcc ttcaccttcc tgcacgactt
     3721 cgaggtgttc gaggagaccc cgctggatct ctccggcttc accgatccgg aggaactggc
     3781 gcagttcctc tgggggccgc cggacaccgc ccccggcgcc tgacgccccg ggccgcccgg
     3841 cgccgccccc ggcccccggc ggccggccgc ccggagcccc gcccgcgctc gggagccccg
     3901 ggcccgcgcc gaagcccgct gctgcgagcc cggagcgggc cggccgacgg cggtgcgggc
     3961 ccggcggcgg acgctcagca gcggcgggcg tgaaaggccc tggcatcctc gatcatctcc
     4021 tccagggtgg tcggccgagc ttccatccca gctcggcaag gatcaacgag gacagtcgca
     4081 cgacgaagtt cttctggatc gcgcccgtgc tggaagcact caacctcgaa gcgtgtggtt
     4141 gcggagccat ctagcaacca cacgaaacat gcgcaacgaa ccgcgcaacg aacaacgcct
     4201 agaactggcc ctagatgagc tgactcgtat cgttggtaaa cctagtttga ccagcatgtt
     4261 ttaactacgt tcggtgagct gtcaacgggg cctgtaacgg cacaacgaac cgtgcaacga
     4321 gagtggccac ggatgccacc acaggcacta caacggagtt cgccacgtac atcaccacaa
     4381 ccaccgattc tggcggtgag ctccccgata ttcagcggaa atggcttggt atcgaccaag
     4441 attcgtagaa ccccgtctcg tctggctggt attcaaaacg ggcgcaacga aacacgcaac
     4501 gagacaggca tggcccaaac cagaaaacta gcgtctacca ggacttttac ctgtccgacc
     4561 cgttgcaacg gaacccccca cggaaccccc gcgacacccg ctccccaatt gcgttagaac
     4621 agcggtggat tgtcggcttc gttgtgggcc ttttgagccg cttcctgttc tgccgcacgc
     4681 tctttcctcg cccgatagcc gagtcgctta acggtgtcca gatgcagccc gaaatgtttg
     4741 gccgtttgcg gccaagagtg gccctcgtcg tcgtgatagg cgcggatgcg ttcgcggcgt
     4801 gcagcctgct cggcgagcca ctcgctgcgt tcctgcgcca cgagccggac gacgtggcgt
     4861 tcggatagtc cggtgattcg agcgccttcg gcggcggtca cgcgccgctt tttgcggaca
     4921 gtcggctgcc ggttgtagcc gtcgctgtag ccgtcgctca tagcaatgcc tccatggctg
     4981 acgcggactt tgcgcgccgc gcaactgtgc tcgccgccgt gcgcgctgct gcgcccttcc
     5041 gcgagatggc cgactggcgc gcactgagtg tggcctcgta gaccacgatc ccgtccgccc
     5101 aaatgcgcga cttggttgtg atccaacgcc aaatgctgtt ggcgatggcg cggacctcgc
     5161 tgtccggtag cggtccggga cacacgtcgt tgcacgggaa ttcggcgttt cgcgcgtggc
     5221 actcggcata gatcgcgcgg ccgagtccgt ccacgttccg ggtcggcagg tagatccgca
     5281 tgagggcggg acgataggcc cacaacctga cggaatcgaa cagtgcgcaa ttccgcccta
     5341 gcggcgtcgg agccgctttg tacgtggtct gctgacgcca gcgcggcggt ggcatgttcg
     5401 cgccgagctc ggcctcgatg tggctgagtg tgtagagatc tgagtggagc cattccgttt
     5461 cccaggcgat gtggccgggg tttttggtca tgaggcctga gtaactgcgg tcgccgtcga
     5521 cggcgcgccg aaggccttcg gcgcacgccg ccatgtatgc gagcggctta cgccgcgcgt
     5581 attcggtgcg tggaacaggg gcgttgagtg cccacactgc gtgtgcgtgg ccgttggcgc
     5641 gattgcccac gatcgcgttg ggcagcggat gggacccccg ggcgctgagc gctcggagcg
     5701 ctgcgtctgg atggtctacg tccacgacca gcaggtttgc cagcgctgtt gggttcgcct
     5761 cgatgtaccg gcggcctagg gccgacgcgc ggctttggcg gtagatcccc tcgagcagat
     5821 cgtcgcttgc cagcggccag tacggcagcc agagctgctc aaattcgtcg gcgacgtggc
     5881 tcacgcttgg tagtagacca cgattaatca ccggtgtatg gtccgacacg agctccaagt
     5941 cagatatttc gctgaggggc caccccacaa ctgcacactc ccccgctctc ccgtcgagcc
     6001 ctggtggtgg aacaccagcg acagccgagc acccccaacc acctgtacca accaggagga
     6061 acacatgcgt cgtttcgagg acgtttccgg gccgctgaga gccgctgtgg cggccgtaca
     6121 cgccgcctta gacccgttag accccctgcc gcctgaatgc gcgggtacga gccacacagc
     6181 gcccgaactt acggagctgg tgggctcacc tggctttatg gcgtacgaat cggctgtgtg
     6241 cgacctgttg ggcgaggtga ggtacgcgct actcacgctg gcaagggcga cagagccgcc
     6301 ccaccgagcc cgcacggccg cgcgcggtgt caacaaccgg gtgagtcgtg cacaccagca
     6361 ggtgttcgag gcttggctcg aagtgcagga catcgtggcg aacgccgccc gatgagccgc
     6421 gccttacgct ggctgccagc cgttcgcggg ctggttggtg cagcgcgtcg agcggttaga
     6481 ggccctgcgg tgttccacca ccgcagggcc tcgccctttt taaggctgaa tttgcttgtc
     6541 tccgaatcca actggcttgt ccaagggtgt atctacgctt aatccaaagt tcaaacgagg
     6601 ggattacaca tgaccaactt cgat
//
