LOCUS       sequence-178199-        2484 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  gRNA for cleavage at dpy-10(cn64) locus in C elegans.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2484)
  AUTHORS   Arribere JA, Bell RT, Fu BX, Artiles KL, Hartman PS, Fire AZ
  TITLE     Efficient Marker-Free Recovery of Custom Genetic Modifications with 
            CRISPR/Cas9 in Caenorhabditis elegans.
  JOURNAL   Genetics. 2014 Aug 26. pii: genetics.114.169730.
  PUBMED    25161212
REFERENCE   2  (bases 1 to 2484)
  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: "Genetics. 
            2014 Aug 26. pii: genetics.114.169730."
COMMENT     Sequence Label: pJA58
FEATURES             Location/Qualifiers
     source          1..2484
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     50..66
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     50..66
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     misc_RNA        complement(209..284)
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     promoter        complement(305..654)
                     /label=CeU6 promoter
                     /note="promoter for the C. elegans R07E5.16 U6 snRNA gene 
                     (Dickinson et al., 2013)"
     primer_bind     complement(829..846)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(829..845)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     rep_origin      886..1473
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1374..1393
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(1485..1512)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(1536..2351)
                     /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     2259..2278
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     promoter        complement(2352..2454)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     terminator      complement(join(2455..2484,1..2))
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
ORIGIN
        1 ctttctgcta tggaggtcag gtatgattta aatggtcagt attgagcctc aggaaacagc
       61 tatgacatca agctgactag ataatctagc tgatcgtgga ccgatcatac gtataatgcc
      121 gtaagatcac gggtcgcagc acagctcgcg gtccagtagt gatcgacact gctcgatccg
      181 ctcgcaccgc tagcaaattt cacaaaaagc accgactcgg tgccactttt tcaagttgat
      241 aacggactag ccttatttta acttgctatt tctagctcta aaacctcgtg gtgcctatgg
      301 tagcaagaca tctcgcaata ggaggtgttt atataggggc aacacaagag gcggcagaca
      361 tctcgtgagt ttgacagagt agtataaaga gtgtgtgtga gagggtgtag gcatttcata
      421 acatattttt tgtggaataa ttttggaaaa aaggtgtttt tttaataata gcttttataa
      481 gagtttctca aaaattgcat gctttttttg aaaaggttga caaattatag tagtactaaa
      541 gttgctattt tcaagtctca gcagaaaatt cacaactaaa aataaaattt gcaaaacaga
      601 gtgtgcgcct ttaaggtgta cagtttttta ttcctttatt gctagttttt tttgaagctt
      661 ggatcgacga gagcagcgcg actggatctg tcgcccgtct caaacgcaac cctccggcgg
      721 tcgcatatca ttcaggacga gcctcagact ccagcgtaac tggactgcaa tcaactcact
      781 ggctcacctt ccggtccacg atcagctaga atcaagctga ctagataaac tggccgtcgt
      841 tttacacggg tgggcctttc ttcggtagaa aatcaaagga tcttcttgag atcctttttt
      901 tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt
      961 gccggatcaa gagctaccaa ctctttttcc gaggtaactg gcttcagcag agcgcagata
     1021 ccaaatactg ttcttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca
     1081 ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag
     1141 tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc
     1201 tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga
     1261 tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg
     1321 tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac
     1381 gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagca tcgatttttg
     1441 tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcaga aaggcccacc
     1501 cgaaggtgag ccaggtgatt acatttaggt cctcattaga aaaactcatc gagcatcaag
     1561 tgaaactgca atttattcat atcaggatta tcaataccat atttttgaaa aagccgtttc
     1621 tgtaatgaag gagaaaactc accgaggcag ttccatagga tggcaagatc ctggtatcgg
     1681 tctgcgattc cgactcgtcc aacatcaata caacctatta atttcccctc gtcaaaaata
     1741 aggttatcaa gtgagaaatc accatgagtg acgactgaat ccggtgagaa tggcaagagt
     1801 ttatgcattt ctttccagac ttgttcaaca ggccagccat tacgctcgtc atcaaaatca
     1861 ctcgcaccaa ccaaaccgtt attcattcgt gattgcgcct gagcgagacg aaatacgcga
     1921 tcgccgttaa aaggacaatt acaaacagga atcgaatgca accggcgcag gaacactgcc
     1981 agcgcatcaa caatattttc acctgaatca ggatattctt ctaatacctg gaatgctgtt
     2041 ttccctggga tcgcagtggt gagtaaccat gcatcatcag gagtacggat aaaatgcttg
     2101 atggtcggaa gaggcataaa ttccgtcagc cagtttagcc tgaccatctc atctgtaaca
     2161 tcattggcaa cgctaccttt gccatgtttc agaaacaact ctggcgcatc gggcttccca
     2221 tacaatcgat agattgtcgc acctgattgc ccgacattat cgcgagccca tttataccca
     2281 tataaatcag catccatgtt ggaatttaat cgcggcttca agcaagacgt ttcccgttga
     2341 atatggctca ttttagcttc cttagctcct gaaaatctcg ataactcaaa aaatacgccc
     2401 ggtagtgatc ttatttcatt atggtgaaag ttggaacctc ttacgtgccg atcaagtcaa
     2461 aagcctccgg tcggaggctt ttga
//
