LOCUS       sequence-381186-        2494 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  sgRNA targeting ChrV_oxTi365 locus of the C. elegans genome.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2494)
  AUTHORS   Kurashina M, Mizumoto K
  TITLE     Targeting endogenous proteins for spatial and temporal knockdown 
            using auxin-inducible degron in Caenorhabditis elegans.
  JOURNAL   STAR Protoc. 2023 Jan 13;4(1):102028. doi: 
            10.1016/j.xpro.2022.102028.
  PUBMED    36640369
REFERENCE   2  (bases 1 to 2494)
  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: "STAR 
            Protoc."; date: "2023-01-13"; volume: "4(1):102028. doi"; pages: " 
            10.1016/j.xpro.2022.102028"
COMMENT     Sequence Label: pTK73.ChrV_oxTi365.gRNA
FEATURES             Location/Qualifiers
     source          1..2494
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     3..20
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     4..20
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        195..544
                     /label=CeU6 promoter
                     /note="promoter for the C. elegans R07E5.16 U6 snRNA gene 
                     (Dickinson et al., 2013)"
     primer_bind     complement(793..809)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(793..809)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     terminator      857..888
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
     promoter        889..991
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             992..1807
                     /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="MSHIQRETSCLKPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNGDRVFRLAQAQSRMNNGLVGA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     complement(1065..1084)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     terminator      1831..1858
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     rep_origin      complement(1870..2457)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1950..1969)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cgtgtaaaac gacggccagt ttatctagtc agcttgattc tagctgatcg tggaccggaa
       61 ggtgagccag tgagttgatt gcagtccagt tacgctggag tctgaggctc gtcctgaatg
      121 atatgcgacc gccggagggt tgcgtttgag acgggcgaca gatccagtcg cgctgctctc
      181 gtcgatccaa gcttcaaaaa aaactagcaa taaaggaata aaaaactgta caccttaaag
      241 gcgcacactc tgttttgcaa attttatttt tagttgtgaa ttttctgctg agacttgaaa
      301 atagcaactt tagtactact ataatttgtc aaccttttca aaaaaagcat gcaatttttg
      361 agaaactctt ataaaagcta ttattaaaaa aacacctttt ttccaaaatt attccacaaa
      421 aaatatgtta tgaaatgcct acaccctctc acacacactc tttatactac tctgtcaaac
      481 tcacgagatg tctgccgcct cttgtgttgc ccctatataa acacctccta ttgcgagatg
      541 tcttgcggtc gcggtactcc aaatgtttaa gagctatgct ggaaacagca tagcaagttt
      601 aaataaggct agtccgttat caacttgaaa aagtggcacc gagtcggtgc tttttgtgaa
      661 atttgctagc ggtgcgagcg gatcgagcag tgtcgatcac tactggaccg cgagctgtgc
      721 tgcgacccgt gatcttacgg cattatacgt atgatcggtc cacgatcagc tagattatct
      781 agtcagcttg atgtcatagc tgtttcctga ggctcaatac tgaccattta aatcatacct
      841 gacctccata gcagaaagtc aaaagcctcc gaccggaggc ttttgacttg atcggcacgt
      901 aagaggttcc aactttcacc ataatgaaat aagatcacta ccgggcgtat tttttgagtt
      961 atcgagattt tcaggagcta aggaagctaa aatgagccat attcaacggg aaacgtcttg
     1021 cttgaagccg cgattaaatt ccaacatgga tgctgattta tatgggtata aatgggctcg
     1081 cgataatgtc gggcaatcag gtgcgacaat ctatcgattg tatgggaagc ccgatgcgcc
     1141 agagttgttt ctgaaacatg gcaaaggtag cgttgccaat gatgttacag atgagatggt
     1201 caggctaaac tggctgacgg aatttatgcc tcttccgacc atcaagcatt ttatccgtac
     1261 tcctgatgat gcatggttac tcaccactgc gatcccaggg aaaacagcat tccaggtatt
     1321 agaagaatat cctgattcag gtgaaaatat tgttgatgcg ctggcagtgt tcctgcgccg
     1381 gttgcattcg attcctgttt gtaattgtcc ttttaacggc gatcgcgtat ttcgtctcgc
     1441 tcaggcgcaa tcacgaatga ataacggttt ggttggtgcg agtgattttg atgacgagcg
     1501 taatggctgg cctgttgaac aagtctggaa agaaatgcat aaactcttgc cattctcacc
     1561 ggattcagtc gtcactcatg gtgatttctc acttgataac cttatttttg acgaggggaa
     1621 attaataggt tgtattgatg ttggacgagt cggaatcgca gaccgatacc aggatcttgc
     1681 catcctatgg aactgcctcg gtgagttttc tccttcatta cagaaacggc tttttcaaaa
     1741 atatggtatt gataatcctg atatgaataa attgcagttt cacttgatgc tcgatgagtt
     1801 tttctaatga ggacctaaat gtaatcacct ggctcacctt cgggtgggcc tttctgcgtt
     1861 gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc gatgctcaag
     1921 tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc
     1981 cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc
     2041 ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt
     2101 cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt
     2161 atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc
     2221 agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa
     2281 gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa
     2341 gccagttacc tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt
     2401 agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa
     2461 gatcctttga ttttctaccg aagaaaggcc cacc
//
