LOCUS       sequence-309427-        3493 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 3493)
  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 3493)
  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: pJW1850
FEATURES             Location/Qualifiers
     source          1..3493
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(25..47)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     147..166
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     360..382
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     374..391
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     375..391
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        398..897
                     /label=CeU6 promoter
                     /note="promoter for the C. elegans K09B11.12 U6 snRNA gene 
                     (Friedland et al., 2013)"
     misc_feature    1011..1247
                     /label=3' CeU6
                     /note="region downstream of the C. elegans K09B11.12 U6 
                     snRNA gene"
     primer_bind     complement(1266..1282)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1266..1282)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1279..1301)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1290..1306
                     /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(1314..1320,1321..1338,1339..1344))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1359..1380
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1499..1516)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1670..2258)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1750..1769)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2429..3220,3221..3289))
                     /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     3052..3071
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3290..3394)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3462..3480
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 gcgtttcggt gatgacggtg aaaacctctg acacatgcag ctcccggaga cggtcacagc
       61 ttgtctgtaa gcggatgccg ggagcagaca agcccgtcag ggcgcgtcag cgggtgttgg
      121 cgggtgtcgg ggctggctta actatgcggc atcagagcag attgtactga gagtgcacca
      181 tatgcggtgt gaaataccgc acagatgcgt aaggagaaaa taccgcatca ggcgccattc
      241 gccattcagg ctgcgcaact gttgggaagg gcgatcggtg cgggcctctt cgctattacg
      301 ccagctggcg aaagggggat gtgctgcaag gcgattaagt tgggtaacgc cagggttttc
      361 ccagtcacga cgttgtaaaa cgacggccag tgaattcctc caagaactcg tacaaaaatg
      421 ctctgaagta ggtctcgaga tcaatactgg gaagacgaaa gtcttgcgaa accgattcgc
      481 tgaccccagt aaagtctact tcggtagccc ttcccccacc acccagctcg acgacgtcga
      541 cgagtacatc tacctcggtc gtcaaatcaa cgcccaaaac aacttgatgc cggaaatcca
      601 ccgaagacgt cgagcagcct gggctgcatt caatggaatc aagaatgcca ccgactccat
      661 caccgacaag aagattcgtg cgaatctgtt cgactcaatt gtccttccag cgctcaccta
      721 cgtttcagaa gcctggacat tcaccaaagc tctatccgaa cgagtacgaa tcacacatgc
      781 ctcccccaca aattattgat ctacatcccc atttggtata gtgtcttaca ggactctcaa
      841 tatgtcagct gcatgataaa tggtccctat ataattgaat tgcaaatcta aatgtttgat
      901 atcagtctgt ttcgtaagtt taagagctat gctggaaaca gcatagcaag tttaaataag
      961 gctagtccgt tatcaacttg aaaaagtggc accgagtcgg tgcttttttt caataatatt
     1021 ctagatatcc ctttttgaaa ctaaaatgaa atactctaaa tttttgagtt caatgtttca
     1081 actaatcttg gttaaaggta aattcccgga gggacgattt taaagttgcc agagttgatt
     1141 aaattggtcc aaaaaatttt attaggaaac tcgaaaaaaa attgattttt tttgacaata
     1201 ttccaaagaa aattacaaaa tctcgacgcc aaacatagtc ggctgtgaag cttggcgtaa
     1261 tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc acacaacata
     1321 cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta actcacatta
     1381 attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa
     1441 tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcata tgcgcttcct
     1501 cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa
     1561 aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa
     1621 aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc
     1681 tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga
     1741 caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc
     1801 cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt
     1861 ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct
     1921 gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg
     1981 agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta
     2041 gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct
     2101 acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa
     2161 gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt
     2221 gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta
     2281 cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat
     2341 caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa
     2401 gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct
     2461 cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta
     2521 cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct
     2581 caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg
     2641 gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa
     2701 gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt
     2761 cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta
     2821 catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca
     2881 gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta
     2941 ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct
     3001 gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg
     3061 cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac
     3121 tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact
     3181 gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa
     3241 atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt
     3301 ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat
     3361 gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg
     3421 acgtctaaga aaccattatt atcatgacat taacctataa aaataggcgt atcacgaggc
     3481 cctttcgtct cgc
//
