LOCUS       sequence-308857-        3494 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 3494)
  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 3494)
  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: pJW1849
FEATURES             Location/Qualifiers
     source          1..3494
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(37..59)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     159..178
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     372..394
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     386..403
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     387..403
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        410..909
                     /label=CeU6 promoter
                     /note="promoter for the C. elegans K09B11.12 U6 snRNA gene 
                     (Friedland et al., 2013)"
     misc_feature    1024..1260
                     /label=3' CeU6
                     /note="region downstream of the C. elegans K09B11.12 U6 
                     snRNA gene"
     primer_bind     complement(1279..1295)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1279..1295)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1292..1314)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1303..1319
                     /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(1327..1333,1334..1351,1352..1357))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1372..1393
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1512..1529)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1683..2271)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1763..1782)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2442..3233,3234..3302))
                     /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     3065..3084
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3303..3407)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3475..3493
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 ctttcgtctc gcgcgtttcg gtgatgacgg tgaaaacctc tgacacatgc agctcccgga
       61 gacggtcaca gcttgtctgt aagcggatgc cgggagcaga caagcccgtc agggcgcgtc
      121 agcgggtgtt ggcgggtgtc ggggctggct taactatgcg gcatcagagc agattgtact
      181 gagagtgcac catatgcggt gtgaaatacc gcacagatgc gtaaggagaa aataccgcat
      241 caggcgccat tcgccattca ggctgcgcaa ctgttgggaa gggcgatcgg tgcgggcctc
      301 ttcgctatta cgccagctgg cgaaaggggg atgtgctgca aggcgattaa gttgggtaac
      361 gccagggttt tcccagtcac gacgttgtaa aacgacggcc agtgaattcc tccaagaact
      421 cgtacaaaaa tgctctgaag taggtctcga gatcaatact gggaagacga aagtcttgcg
      481 aaaccgattc gctgacccca gtaaagtcta cttcggtagc ccttccccca ccacccagct
      541 cgacgacgtc gacgagtaca tctacctcgg tcgtcaaatc aacgcccaaa acaacttgat
      601 gccggaaatc caccgaagac gtcgagcagc ctgggctgca ttcaatggaa tcaagaatgc
      661 caccgactcc atcaccgaca agaagattcg tgcgaatctg ttcgactcaa ttgtccttcc
      721 agcgctcacc tacgtttcag aagcctggac attcaccaaa gctctatccg aacgagtacg
      781 aatcacacat gcctccccca caaattattg atctacatcc ccatttggta tagtgtctta
      841 caggactctc aatatgtcag ctgcatgata aatggtccct atataattga attgcaaatc
      901 taaatgtttg gaaatcgccg acttgcgagg gtttaagagc tatgctggaa acagcatagc
      961 aagtttaaat aaggctagtc cgttatcaac ttgaaaaagt ggcaccgagt cggtgctttt
     1021 tttcaataat attctagata tccctttttg aaactaaaat gaaatactct aaatttttga
     1081 gttcaatgtt tcaactaatc ttggttaaag gtaaattccc ggagggacga ttttaaagtt
     1141 gccagagttg attaaattgg tccaaaaaat tttattagga aactcgaaaa aaaattgatt
     1201 ttttttgaca atattccaaa gaaaattaca aaatctcgac gccaaacata gtcggctgtg
     1261 aagcttggcg taatcatggt catagctgtt tcctgtgtga aattgttatc cgctcacaat
     1321 tccacacaac atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag
     1381 ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg
     1441 ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc
     1501 atatgcgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta
     1561 tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag
     1621 aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg
     1681 tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg
     1741 tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg
     1801 cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga
     1861 agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc
     1921 tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt
     1981 aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact
     2041 ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg
     2101 cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt
     2161 accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt
     2221 ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct
     2281 ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg
     2341 gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt
     2401 aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt
     2461 gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc
     2521 gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg
     2581 cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc
     2641 gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg
     2701 gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca
     2761 ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga
     2821 tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct
     2881 ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg
     2941 cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca
     3001 accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata
     3061 cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct
     3121 tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact
     3181 cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa
     3241 acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc
     3301 atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga
     3361 tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga
     3421 aaagtgccac ctgacgtcta agaaaccatt attatcatga cattaaccta taaaaatagg
     3481 cgtatcacga ggcc
//
