LOCUS       sequence-95095-e        2507 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  empty vector for gRNA cloning.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2507)
  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 2507)
  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: pRB1017
FEATURES             Location/Qualifiers
     source          1..2507
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     5..22
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     6..22
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        197..546
                     /label=CeU6 promoter
                     /note="promoter for the C. elegans R07E5.16 U6 snRNA gene 
                     (Dickinson et al., 2013)"
     misc_RNA        590..665
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     primer_bind     complement(808..824)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(808..824)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     terminator      872..903
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
     promoter        904..1006
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             1007..1822
                     /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(1080..1099)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     terminator      1846..1873
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     rep_origin      complement(1885..2472)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1965..1984)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cccgtgtaaa acgacggcca gtttatctag tcagcttgat tctagctgat cgtggaccgg
       61 aaggtgagcc agtgagttga ttgcagtcca gttacgctgg agtctgaggc tcgtcctgaa
      121 tgatatgcga ccgccggagg gttgcgtttg agacgggcga cagatccagt cgcgctgctc
      181 tcgtcgatcc aagcttcaaa aaaaactagc aataaaggaa taaaaaactg tacaccttaa
      241 aggcgcacac tctgttttgc aaattttatt tttagttgtg aattttctgc tgagacttga
      301 aaatagcaac tttagtacta ctataatttg tcaacctttt caaaaaaagc atgcaatttt
      361 tgagaaactc ttataaaagc tattattaaa aaaacacctt ttttccaaaa ttattccaca
      421 aaaaatatgt tatgaaatgc ctacaccctc tcacacacac tctttatact actctgtcaa
      481 actcacgaga tgtctgccgc ctcttgtgtt gcccctatat aaacacctcc tattgcgaga
      541 tgtcttggag accggtaacc atggctcgag aaaccggtac tcggtctctg ttttagagct
      601 agaaatagca agttaaaata aggctagtcc gttatcaact tgaaaaagtg gcaccgagtc
      661 ggtgcttttt gtgaaatttg ctagcggtgc gagcggatcg agcagtgtcg atcactactg
      721 gaccgcgagc tgtgctgcga cccgtgatct tacggcatta tacgtatgat cggtccacga
      781 tcagctagat tatctagtca gcttgatgtc atagctgttt cctgaggctc aatactgacc
      841 atttaaatca tacctgacct ccatagcaga aagtcaaaag cctccgaccg gaggcttttg
      901 acttgatcgg cacgtaagag gttccaactt tcaccataat gaaataagat cactaccggg
      961 cgtatttttt gagttatcga gattttcagg agctaaggaa gctaaaatga gccatattca
     1021 acgggaaacg tcttgcttga agccgcgatt aaattccaac atggatgctg atttatatgg
     1081 gtataaatgg gctcgcgata atgtcgggca atcaggtgcg acaatctatc gattgtatgg
     1141 gaagcccgat gcgccagagt tgtttctgaa acatggcaaa ggtagcgttg ccaatgatgt
     1201 tacagatgag atggtcaggc taaactggct gacggaattt atgcctcttc cgaccatcaa
     1261 gcattttatc cgtactcctg atgatgcatg gttactcacc actgcgatcc cagggaaaac
     1321 agcattccag gtattagaag aatatcctga ttcaggtgaa aatattgttg atgcgctggc
     1381 agtgttcctg cgccggttgc attcgattcc tgtttgtaat tgtcctttta acggcgatcg
     1441 cgtatttcgt ctcgctcagg cgcaatcacg aatgaataac ggtttggttg gtgcgagtga
     1501 ttttgatgac gagcgtaatg gctggcctgt tgaacaagtc tggaaagaaa tgcataaact
     1561 cttgccattc tcaccggatt cagtcgtcac tcatggtgat ttctcacttg ataaccttat
     1621 ttttgacgag gggaaattaa taggttgtat tgatgttgga cgagtcggaa tcgcagaccg
     1681 ataccaggat cttgccatcc tatggaactg cctcggtgag ttttctcctt cattacagaa
     1741 acggcttttt caaaaatatg gtattgataa tcctgatatg aataaattgc agtttcactt
     1801 gatgctcgat gagtttttct aatgaggacc taaatgtaat cacctggctc accttcgggt
     1861 gggcctttct gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa
     1921 aaatcgatgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt
     1981 tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct
     2041 gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct
     2101 cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc
     2161 cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt
     2221 atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc
     2281 tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat
     2341 ctgcgctctg ctgaagccag ttacctcgga aaaagagttg gtagctcttg atccggcaaa
     2401 caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa
     2461 aaaggatctc aagaagatcc tttgattttc taccgaagaa aggccca
//
