LOCUS       sequence-71633-e        3423 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  to drive the sgRNA expression under RNase P non-coding RNA promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3423)
  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 3423)
  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 rpr-1 promoter
FEATURES             Location/Qualifiers
     source          1..3423
                     /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"
     protein_bind    420..440
                     /gene="mutant version of attB"
                     /label=attB6
                     /bound_moiety="BP Clonase(TM)"
                     /note="core recombination site for the Gateway(R) BP 
                     reaction"
     misc_RNA        913..988
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     primer_bind     complement(1566..1583)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1676..2264)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1756..1775)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2435..3226,3227..3295))
                     /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     3058..3077
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3296..3400)
                     /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 gcgcgccacg atttcggcta aaaaatagcg aaaaaacatc
      421 aaatttgtat taaaaagaag cagttgaaat tttgagtgag gctcagagac tacaaactac
      481 aaaaaggctc agcctcaacc aatttttagt gttaaaaatt tgattttttc agttaaaaac
      541 gatggttttt gatgcttttt acctatttta cagatagaaa attttaaatt tcgcaaaaat
      601 ctttaaaaat aaactttttt gtttggtttt cgctcgaaaa tagctttaaa tttgcagttt
      661 ttctctcaaa aactctaaat ttcatgcgtt ccacgtggct ttctatcaat ttaaatacta
      721 atattaatat tttcttcaaa atctgccacg ttacagcctg ccatagcctt ctgaaactct
      781 acaattcccc tctaatcact agcgccgcct ccccgcacac tcaacacagc gagcggcgga
      841 acccgcgggg tgtcggccgc tcggacacgc tcccgtatat aacgacgcgg cgcgcgtcaa
      901 gttgtggaat tcgttttaga gctagaaata gcaagttaaa ataaggctag tccgttatca
      961 acttgaaaaa gtggcaccga gtcggtgctt ttgtgggctt caggctttct gttgggccgt
     1021 tttaggccaa atgatagaaa gccacgtgga aatcatgaat ttaaagctat tttcgagcga
     1081 aaatcagaaa aaaagtttat tttttaaaga tttttgctga attttacatt ttctaaatgt
     1141 aaaataggtg aaaagcatca aaaaccatag tttttaactg aaaaaatcaa atttttaaca
     1201 ctaaaaattg gttgaggctg atcctttttg tagtttgtag tctctgagcc tcactcaaaa
     1261 tttcaaaagt tctttttgat acaaatttga tgatttttcg ctatttttct agccgaaatc
     1321 gtatttttca acattttctt aacaaaatta ggatttgctc aaaattgata taacattatt
     1381 taaaagtacc gttaaagttg taccaaaaat agtggaaatt tatttttttt tattaattaa
     1441 tggagcacaa gactggcctc atgggccttc cgctcactgc ccgctttcca gtcgggaaac
     1501 ctgtcgtgcc agctgcatta acatggtcat agctgtttcc ttgcgtattg ggcgctctcc
     1561 gcttcctcgc tcactgactc gctgcgctcg gtcgttcggg taaagcctgg ggtgcctaat
     1621 gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc
     1681 ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa
     1741 acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc
     1801 ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg
     1861 cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc
     1921 tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc
     1981 gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca
     2041 ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact
     2101 acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg
     2161 gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt
     2221 ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct
     2281 tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga
     2341 gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa
     2401 tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac
     2461 ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga
     2521 taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagaac
     2581 cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca
     2641 gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta
     2701 gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg
     2761 tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc
     2821 gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg
     2881 ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt
     2941 ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt
     3001 cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata
     3061 ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc
     3121 gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac
     3181 ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa
     3241 ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct
     3301 tcctttttca atattattga agcatttatc agggttattg tctcatgagc ggatacatat
     3361 ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc
     3421 cac
//
