LOCUS       sequence-382110-        2494 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  sgRNA targeting ChrII_ttTi5605 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.ChrII_ttTi5605.sgRNA
FEATURES             Location/Qualifiers
     source          1..2494
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     8..25
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     9..25
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        200..549
                     /label=CeU6 promoter
                     /note="promoter for the C. elegans R07E5.16 U6 snRNA gene 
                     (Dickinson et al., 2013)"
     primer_bind     complement(798..814)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(798..814)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     terminator      862..893
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
     promoter        894..996
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             997..1812
                     /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(1070..1089)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     terminator      1836..1863
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     rep_origin      complement(1875..2462)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1955..1974)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ccacccgtgt aaaacgacgg ccagtttatc tagtcagctt gattctagct gatcgtggac
       61 cggaaggtga gccagtgagt tgattgcagt ccagttacgc tggagtctga ggctcgtcct
      121 gaatgatatg cgaccgccgg agggttgcgt ttgagacggg cgacagatcc agtcgcgctg
      181 ctctcgtcga tccaagcttc aaaaaaaact agcaataaag gaataaaaaa ctgtacacct
      241 taaaggcgca cactctgttt tgcaaatttt atttttagtt gtgaattttc tgctgagact
      301 tgaaaatagc aactttagta ctactataat ttgtcaacct tttcaaaaaa agcatgcaat
      361 ttttgagaaa ctcttataaa agctattatt aaaaaaacac cttttttcca aaattattcc
      421 acaaaaaata tgttatgaaa tgcctacacc ctctcacaca cactctttat actactctgt
      481 caaactcacg agatgtctgc cgcctcttgt gttgccccta tataaacacc tcctattgcg
      541 agatgtcttg atatcagtct gtttcgtaag tttaagagct atgctggaaa cagcatagca
      601 agtttaaata aggctagtcc gttatcaact tgaaaaagtg gcaccgagtc ggtgcttttt
      661 gtgaaatttg ctagcggtgc gagcggatcg agcagtgtcg atcactactg gaccgcgagc
      721 tgtgctgcga cccgtgatct tacggcatta tacgtatgat cggtccacga tcagctagat
      781 tatctagtca gcttgatgtc atagctgttt cctgaggctc aatactgacc atttaaatca
      841 tacctgacct ccatagcaga aagtcaaaag cctccgaccg gaggcttttg acttgatcgg
      901 cacgtaagag gttccaactt tcaccataat gaaataagat cactaccggg cgtatttttt
      961 gagttatcga gattttcagg agctaaggaa gctaaaatga gccatattca acgggaaacg
     1021 tcttgcttga agccgcgatt aaattccaac atggatgctg atttatatgg gtataaatgg
     1081 gctcgcgata atgtcgggca atcaggtgcg acaatctatc gattgtatgg gaagcccgat
     1141 gcgccagagt tgtttctgaa acatggcaaa ggtagcgttg ccaatgatgt tacagatgag
     1201 atggtcaggc taaactggct gacggaattt atgcctcttc cgaccatcaa gcattttatc
     1261 cgtactcctg atgatgcatg gttactcacc actgcgatcc cagggaaaac agcattccag
     1321 gtattagaag aatatcctga ttcaggtgaa aatattgttg atgcgctggc agtgttcctg
     1381 cgccggttgc attcgattcc tgtttgtaat tgtcctttta acggcgatcg cgtatttcgt
     1441 ctcgctcagg cgcaatcacg aatgaataac ggtttggttg gtgcgagtga ttttgatgac
     1501 gagcgtaatg gctggcctgt tgaacaagtc tggaaagaaa tgcataaact cttgccattc
     1561 tcaccggatt cagtcgtcac tcatggtgat ttctcacttg ataaccttat ttttgacgag
     1621 gggaaattaa taggttgtat tgatgttgga cgagtcggaa tcgcagaccg ataccaggat
     1681 cttgccatcc tatggaactg cctcggtgag ttttctcctt cattacagaa acggcttttt
     1741 caaaaatatg gtattgataa tcctgatatg aataaattgc agtttcactt gatgctcgat
     1801 gagtttttct aatgaggacc taaatgtaat cacctggctc accttcgggt gggcctttct
     1861 gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgatgc
     1921 tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga
     1981 agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt
     2041 ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg
     2101 taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc
     2161 gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg
     2221 gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc
     2281 ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg
     2341 ctgaagccag ttacctcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg
     2401 ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc
     2461 aagaagatcc tttgattttc taccgaagaa aggc
//
