LOCUS       sequence-111541-        3319 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  sgRNA targeting dpy-10 (Arribere et al., 2014) expressed from a C. 
            elegans U6 promoter (Friedland et al., 2013), in a ""flipped and 
            extended"" sgRNA backbone (Chen et al., 2013)"".
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3319)
  AUTHORS   Katic I, Xu L, Ciosk R
  TITLE     CRISPR/Cas9 Genome Editing in Caenorhabditis elegans: Evaluation of 
            Templates for Homology-Mediated Repair and Knock-Ins by 
            Homology-Independent DNA Repair.
  JOURNAL   G3 (Bethesda). 2015 Jun 3. pii: g3.115.019273. doi: 
            10.1534/g3.115.019273.
  PUBMED    26044730
REFERENCE   2  (bases 1 to 3319)
  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: "G3 
            (Bethesda). 2015 Jun 3. pii: g3.115.019273. doi: 
            10.1534/g3.115.019273."
COMMENT     Sequence Label: pIK208
FEATURES             Location/Qualifiers
     source          1..3319
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(29..51)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     151..170
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     364..386
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     378..395
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     379..395
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        397..896
                     /label=CeU6 promoter
                     /note="promoter for the C. elegans K09B11.12 U6 snRNA gene 
                     (Friedland et al., 2013)"
     primer_bind     complement(1098..1114)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1098..1114)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1111..1133)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1122..1138
                     /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(1146..1152,1153..1170,1171..1176))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1191..1212
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1329..1346)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1500..2088)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1580..1599)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2259..3050,3051..3119))
                     /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     2882..2901
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3120..3224)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3292..3310
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca
       61 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg
      121 ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc
      181 accatatgcg gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc atcaggcgcc
      241 attcgccatt caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat
      301 tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta acgccagggt
      361 tttcccagtc acgacgttgt aaaacgacgg ccagtgctcc aagaactcgt acaaaaatgc
      421 tctgaagtag gtctcgagat caatactggg aagacgaaag tcttgcgaaa ccgattcgct
      481 gaccccagta aagtctactt cggtagccct tcccccacca cccagctcga cgacgtcgac
      541 gagtacatct acctcggtcg tcaaatcaac gcccaaaaca acttgatgcc ggaaatccac
      601 cgaagacgtc gagcagcctg ggctgcattc aatggaatca agaatgccac cgactccatc
      661 accgacaaga agattcgtgc gaatctgttc gactcaattg tccttccagc gctcacctac
      721 gtttcagaag cctggacatt caccaaagct ctatccgaac gagtacgaat cacacatgcc
      781 tcccccacaa attattgatc tacatcccca tttggtatag tgtcttacag gactctcaat
      841 atgtcagctg catgataaat ggtccctata taattgaatt gcaaatctaa atgtttgcta
      901 ccataggcac cacgaggttt aagagctatg ctggaaacag catagcaagt ttaaataagg
      961 ctagtccgtt atcaacttga aaaagtggca ccgagtcggt gctttttttc gagctcggta
     1021 cctcgcgaat gcatctagat atcggatccc gggcccgtcg actgcagagg cctgcatgca
     1081 agcttggcgt aatcatggtc atagctgttt cctgtgtgaa attgttatcc gctcacaatt
     1141 ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta atgagtgagc
     1201 taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc
     1261 cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct
     1321 tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca
     1381 gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac
     1441 atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt
     1501 ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg
     1561 cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc
     1621 tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc
     1681 gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc
     1741 aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac
     1801 tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt
     1861 aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct
     1921 aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc
     1981 ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt
     2041 ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg
     2101 atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc
     2161 atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa
     2221 tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag
     2281 gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg
     2341 tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga
     2401 gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag
     2461 cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa
     2521 gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc
     2581 atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca
     2641 aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg
     2701 atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat
     2761 aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc
     2821 aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg
     2881 gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg
     2941 gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt
     3001 gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca
     3061 ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata
     3121 ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac
     3181 atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa
     3241 gtgccacctg acgtctaaga aaccattatt atcatgacat taacctataa aaataggcgt
     3301 atcacgaggc cctttcgtc
//
