LOCUS       sequence-71639-e        2960 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  to drive the sgRNA expression under a U6 promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2960)
  AUTHORS   Chen C, Fenk LA, de Bono M
  TITLE     Efficient genome editing in Caenorhabditis elegans by 
            CRISPR-targeted homologous recombination.
  JOURNAL   Nucleic Acids Res. 2013 Sep 5.
  PUBMED    24013562
REFERENCE   2  (bases 1 to 2960)
  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: "Nucleic 
            Acids Res. 2013 Sep 5."
COMMENT     Sequence Label: sgRNA with U6 promoter
FEATURES             Location/Qualifiers
     source          1..2960
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     278..300
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     292..309
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     293..309
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     320..339
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        320..338
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     misc_RNA        699..774
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     primer_bind     complement(1103..1120)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1213..1801)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1293..1312)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1972..2763,2764..2832))
                     /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     2595..2614
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2833..2937)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ctaaattgta agcgttaata ttttgttaaa attcgcgtta aatttttgtt aaatcagctc
       61 attttttaac caataggccg aaatcggcaa aatcccttat aaatcaaaag aatagaccga
      121 gatagggttg agtggccgct acagggcgct cccattcgcc attcaggctg cgcaactgtt
      181 gggaagggcg tttcggtgcg ggcctcttcg ctattacgcc agctggcgaa agggggatgt
      241 gctgcaaggc gattaagttg ggtaacgcca gggttttccc agtcacgacg ttgtaaaacg
      301 acggccagtg agcgcgacgt aatacgactc actatagggc gaattggcgg aaggccgtca
      361 aggccacgtg tcttgtccag gcgcgccacc caaatgtttt tataatttgg caaattacca
      421 aaaatgtgac tggaagcatt aacaaaattt tgaacaatat tcgaggtttc aaaatcatga
      481 tattttagat attttggact gttttgagtc aaattgttaa tgacagcatt ttaagagctc
      541 ccaacacata gtgtttccaa tgttataccc aatcaataat agcaagtcaa taaactacct
      601 ctacactatt tctggtaata ggcgaaacct ctacactgtc agtcactttg taaggtgtgc
      661 ctatgtattt ccataatatt tcatacaaat tgaagcttgt tttagagcta gaaatagcaa
      721 gttaaaataa ggctagtccg ttatcaactt gaaaaagtgg caccgagtcg gtgctttttt
      781 tgactaattt ctggtattat tgtttagaat tgtaagaaaa ttttacattc cagaaaatgt
      841 atccaaaaaa cttacgcaaa ctcggagcgt atagttgcat gaagtgctgg atccgaaata
      901 cgaacgacaa attgttgata ctttgctttc aaactctcgt aataaggcat cacaaagttc
      961 tcaattttat taattaatgg agcacaagac tggcctcatg ggccttccgc tcactgcccg
     1021 ctttccagtc gggaaacctg tcgtgccagc tgcattaaca tggtcatagc tgtttccttg
     1081 cgtattgggc gctctccgct tcctcgctca ctgactcgct gcgctcggtc gttcgggtaa
     1141 agcctggggt gcctaatgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc
     1201 gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc
     1261 aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag
     1321 ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct
     1381 cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta
     1441 ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc
     1501 cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc
     1561 agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt
     1621 gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct
     1681 gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc
     1741 tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca
     1801 agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta
     1861 agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa
     1921 atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg
     1981 cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg
     2041 actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc
     2101 aatgataccg cgagaaccac gctcaccggc tccagattta tcagcaataa accagccagc
     2161 cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa
     2221 ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc
     2281 cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg
     2341 ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc
     2401 cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat
     2461 ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg
     2521 tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc
     2581 ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg
     2641 aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat
     2701 gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg
     2761 gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg
     2821 ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct
     2881 catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac
     2941 atttccccga aaagtgccac
//
