LOCUS       sequence-111540-        3307 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  ""flipped and extended"" sgRNA backbone (Chen et al., 2013) 
            following a C. elegans U6 promoter (Friedland et al., 2013). For 
            Gibson cloning locus-specific sgRNA sequences..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3307)
  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 3307)
  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: pIK198
FEATURES             Location/Qualifiers
     source          1..3307
                     /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(1086..1102)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1086..1102)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1099..1121)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1110..1126
                     /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(1134..1140,1141..1158,1159..1164))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1179..1200
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1317..1334)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1488..2076)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1568..1587)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2247..3038,3039..3107))
                     /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     2870..2889
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3108..3212)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3280..3298
                     /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 atgtttgcgg
      901 ccgcgtttaa gagctatgct ggaaacagca tagcaagttt aaataaggct agtccgttat
      961 caacttgaaa aagtggcacc gagtcggtgc tttttttcga gctcggtacc tcgcgaatgc
     1021 atctagatat cggatcccgg gcccgtcgac tgcagaggcc tgcatgcaag cttggcgtaa
     1081 tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc acacaacata
     1141 cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta actcacatta
     1201 attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa
     1261 tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg
     1321 ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag
     1381 gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa
     1441 ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc
     1501 cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca
     1561 ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg
     1621 accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct
     1681 catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt
     1741 gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag
     1801 tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc
     1861 agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac
     1921 actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga
     1981 gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc
     2041 aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg
     2101 gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca
     2161 aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt
     2221 atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca
     2281 gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg
     2341 atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca
     2401 ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt
     2461 cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt
     2521 agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca
     2581 cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca
     2641 tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga
     2701 agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact
     2761 gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga
     2821 gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg
     2881 ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc
     2941 tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga
     3001 tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat
     3061 gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt
     3121 caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt
     3181 atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac
     3241 gtctaagaaa ccattattat catgacatta acctataaaa ataggcgtat cacgaggccc
     3301 tttcgtc
//
