LOCUS       sequence-318645-        3187 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  sgRNA Target Vector for CRISPR.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3187)
  AUTHORS   Ashley GE, Duong T, Levenson MT, Martinez MAQ, Johnson LC, Hibshman 
            JD, Saeger HN, Palmisano NJ, Doonan R, Martinez-Mendez R, Davidson 
            BR, Zhang W, Ragle JM, Medwig-Kinney TN, Sirota SS, Goldstein B, 
            Matus DQ, Dickinson DJ, Reiner DJ, Ward JD
  TITLE     An expanded auxin-inducible degron toolkit for Caenorhabditis 
            elegans.
  JOURNAL   Genetics. 2021 Mar 31;217(3). pii: 6104563. doi: 
            10.1093/genetics/iyab006.
  PUBMED    33677541
REFERENCE   2  (bases 1 to 3187)
  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.";
            date: "2021-03-31"; volume: "217(3). pii: 6104563. doi"; pages: " 
            10.1093/genetics/iyab006"
COMMENT     Sequence Label: pJW1882
FEATURES             Location/Qualifiers
     source          1..3187
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(39..61)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     161..180
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     374..396
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     388..405
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     389..405
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        412..761
                     /label=CeU6 promoter
                     /note="promoter for the C. elegans R07E5.16 U6 snRNA gene 
                     (Dickinson et al., 2013)"
     primer_bind     complement(974..990)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(974..990)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(987..1009)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    998..1014
                     /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)."
     promoter        complement(join(1022..1028,1029..1046,1047..1052))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1067..1088
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1207..1224)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1378..1966)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1458..1477)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2137..2928,2929..2997))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     2760..2779
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2998..3102)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     join(3170..3187,1)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 ccctttcgtc tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg
       61 gagacggtca cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg
      121 tcagcgggtg ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta
      181 ctgagagtgc accatatgcg gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc
      241 atcaggcgcc attcgccatt caggctgcgc aactgttggg aagggcgatc ggtgcgggcc
      301 tcttcgctat tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta
      361 acgccagggt tttcccagtc acgacgttgt aaaacgacgg ccagtgaatt ccaaaaaaaa
      421 ctagcaataa aggaataaaa aactgtacac cttaaaggcg cacactctgt tttgcaaatt
      481 ttatttttag ttgtgaattt tctgctgaga cctgaaaata gcaactttag tactactata
      541 atttgtcaac cttttcaaaa aaagcatgca atttttgaga aactcttata aaagctatta
      601 ttaaaaaaac accttttttc caaaattatt ccacaaaaaa tatgttatga aatgcctaca
      661 ccctctcaca cacactcttt atactactct gtcaaactca cgagatgtct gccgcctctt
      721 gtgttgcccc tatataaaca cctcctattg cgagatgtct tggaaatcgc cgacttgcga
      781 gggtttaaga gctatgctgg aaacagcata gcaagtttaa ataaggctag tccgttatca
      841 acttgaaaaa gtggcaccga gtcggtgctt ttttttgtga aatttctggc gtaatagcga
      901 agaggcccgc accgatcgcc cttcccaaca gttgcgcagc ctgaatggcg aatggaagct
      961 tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac
     1021 acaacatacg agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac
     1081 tcacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc
     1141 tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcatatg
     1201 cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc
     1261 tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat
     1321 gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt
     1381 ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg
     1441 aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc
     1501 tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt
     1561 ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa
     1621 gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta
     1681 tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa
     1741 caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa
     1801 ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt
     1861 cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt
     1921 ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat
     1981 cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat
     2041 gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc
     2101 aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc
     2161 acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta
     2221 gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga
     2281 cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg
     2341 cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc
     2401 tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat
     2461 cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag
     2521 gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat
     2581 cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa
     2641 ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa
     2701 gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga
     2761 taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg
     2821 gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc
     2881 acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg
     2941 aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact
     3001 cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat
     3061 atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt
     3121 gccacctgac gtctaagaaa ccattattat catgacatta acctataaaa ataggcgtat
     3181 cacgagg
//
