LOCUS       sequence-371484-         998 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  All-in-one editing CRISPR-Cas construct for one-step genome editing 
            of Streptomyces using AsCas12j-2.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 998)
  AUTHORS   Tan LL, Heng E, Zulkarnain N, Leong CY, Ng V, Yang LK, San Seow DC, 
            Peh G, Lim YH, Lokanand K, Kanagasundaram YH, Ng SB, Wong FT
  TITLE     Application of Cas12j for Streptomyces editing and cluster 
            activation
  JOURNAL   bioRxiv 2021.10.28.465406
REFERENCE   2  (bases 1 to 998)
  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: "bioRxiv 
            2021.10.28.465406"
COMMENT     Sequence Label: pCRISPomyces-AsCas12j-2
FEATURES             Location/Qualifiers
     source          1..998
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     terminator      complement(170..204)
                     /label=lambda t0 terminator
                     /note="minimal transcription terminator from phage lambda 
                     (Scholtissek and Grosse, 1987)"
     protein_bind    263..284
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(299..304,305..322,323..329)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    337..353
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     342..364
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     361..377
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     361..377
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             373..669
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSAQLTLTKGNKSWRTSRGGPVPNSPYSESYYARSLAVVL
                     QRRDWENPGVTQLNRLAAHPPFASWRNSEEARTDRPSQQLRSLNGEWDAPCSG"
     primer_bind     396..416
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        398..416
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(463..482)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(464..482)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(492..509)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(492..508)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(501..523)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     promoter        complement(714..998)
                     /label=gapdh promoter
                     /note="glyceraldehyde-3-phosphate dehydrogenase promoter 
                     from Eggerthella lenta"
ORIGIN
        1 tagcatgcat atgaagcttt gttcacattc gaacggtctc tgctttgaca acatgctgtg
       61 cggtgttgta aagtcgtggc caggagaata cgacagcgtg caggactggg ggagttatga
      121 ctagtgtgaa aagtcagtat ccagtcgtgt agttcttatt accttctaga taaaaaacgc
      181 ccggcggcaa ccgagcgttc tgaacaaatc cagatggagt atgtcttcac cggtggaaag
      241 cgggcagtga gcgcaacgca attaatgtga gttagctcac tcattaggca ccccaggctt
      301 tacactttat gcttccggct cgtatgttgt gtggaattgt gagcggataa caatttcaca
      361 caggaaacag ctatgaccat gattacgcca agcgcgcaat taaccctcac taaagggaac
      421 aaaagctgga ggacctcgag ggggggcccg gtacccaatt cgccctatag tgagtcgtat
      481 tacgcgcgct cactggccgt cgttttacaa cgtcgtgact gggaaaaccc tggcgttacc
      541 caacttaatc gccttgcagc acatccccct ttcgccagct ggcgtaatag cgaagaggcc
      601 cgcaccgatc gcccttccca acagttgcgc agcctgaatg gcgaatggga cgcgccctgt
      661 agcggctgag aagacttgtc ccctcgtgag gggcaatcgt tgagcgttcc gacgcgtatc
      721 ccctttcaga tactcgcact aagaattgca ggaacgcccc gatcatagcg gtagccgccc
      781 agatgctgca agccttatct ggcagccgta taaaaaaagc aaccgaacag gccattcaca
      841 gaagtttcac accgctcgcc gaggggctcc gcacccggtg cgcgccgtac agcagcgcgc
      901 ttctcccagc atcggccagt tcgcccgatc cgtccgtgtc gcacagccga cggctgcggt
      961 aaggtgcccg tagacgcacg tccgaccgaa ggagcagc
//
