LOCUS       sequence-142372-         907 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  For cloning and constitutive expression of sgRNAs in E.coli. New 
            sgRNAs are cloned into SapI sites and multiplex sgRNAs are cloned 
            into BsaI sites. Kanamycin resistant with pSC101 origin of rep..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 907)
  AUTHORS   Standage-Beier K, Zhang Q, Wang X
  TITLE     Targeted Large-Scale Deletion of Bacterial Genomes Using 
            CRISPR-Nickases.
  JOURNAL   ACS Synth Biol. 2015 Nov 20;4(11):1217-25. doi: 
            10.1021/acssynbio.5b00132. Epub 2015 Oct 25.
  PUBMED    26451892
REFERENCE   2  (bases 1 to 907)
  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"; doi: 
            "10.1021/acssynbio.5b00132"; journalName: "ACS Synth Biol"; date: 
            "2015-11-20- 20"; volume: "4"; issue: "11"; pages: "1217-25"
COMMENT     Sequence Label: pSG4K5
FEATURES             Location/Qualifiers
     source          1..907
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     promoter        80..114
                     /label=J23119 promoter
                     /note="bacterial promoter (Registry of Standard Biological 
                     Parts BBa_J23119)"
     misc_RNA        137..212
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     terminator      313..344
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
ORIGIN
        1 gaaaggccca cccgtgaagg tgagccagtg agttgattgc tacgtaatta gttagttagc
       61 ccttagtgac tcgaattcgt tgacagctag ctcagtccta ggtataatgc tagctagaag
      121 agcgacaggc tcttctgttt tagagctaga aatagcaagt taaaataagg ctagtccgtt
      181 atcaacttga aaaagtggca ccgagtcggt gctttttttc tagataattg gagaccgagc
      241 tggtctctct aggatgtgca ctagtagagc tctgcaggag tcactaaggg ttagttagtt
      301 agattagcag aaagtcaaaa gcctccgacc ggaggctttt gactaaaact tcccttgggg
      361 ttatcattgg ggctcactca aaggcggtaa tcagataaaa aaaatcctta gctttcgcta
      421 aggatgattt ctgctagtat tattagaaaa actcatcgag catcaaatga aactgcaatt
      481 tattcatatc aggattatca ataccatatt tttgaaaaag ccgtttctgt aatgaaggag
      541 aaaactcacc gaggcagttc caaagaatgg caaggtcctg gtaacggtct gcgattccga
      601 cccgtccaac atcaatacaa cctattaatt tcccctcgtc aaaaataagg ttatcaagtg
      661 agaaatcacc atgagtgacg actgaatccg gtgagaatgg caagagcttg tgcatttctt
      721 tccagacttg ttcaacaggc cagccattac gctcgtcatc aaaatcactc gcatcaacca
      781 aaccgttatt catgcgtgat tgcgcctgag caagacgaaa tacacgatcg ctgttaaaag
      841 gacaattaca aacaggaatc gaatgtaacc ggcgcaggaa cacggccagc gcatcaacaa
      901 tattttc
//
