LOCUS       sequence-370712-        3484 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  dpy-10 sgRNA expression vector.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3484)
  AUTHORS   Schwartz ML, Davis MW, Rich MS, Jorgensen EM
  TITLE     High-efficiency CRISPR gene editing in C. elegans using Cas9 
            integrated into the genome.
  JOURNAL   PLoS Genet. 2021 Nov 8;17(11):e1009755. doi: 
            10.1371/journal.pgen.1009755. eCollection 2021 Nov.
  PUBMED    34748534
REFERENCE   2  (bases 1 to 3484)
  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: "PLoS 
            Genet."; date: "2021-11-8"; volume: "17(11):e1009755. doi"; pages: "
            10.1371/journal.pgen.1009755. eCollection 2021 Nov"
COMMENT     Sequence Label: pMLS597
FEATURES             Location/Qualifiers
     source          1..3484
                     /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        402..901
                     /label=CeU6 promoter
                     /note="promoter for the C. elegans K09B11.12 U6 snRNA gene 
                     (Friedland et al., 2013)"
     misc_RNA        923..998
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     misc_feature    1006..1242
                     /label=3' CeU6
                     /note="region downstream of the C. elegans K09B11.12 U6 
                     snRNA gene"
     primer_bind     complement(1261..1277)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1261..1277)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1274..1296)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1285..1301
                     /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(1309..1315,1316..1333,1334..1339))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1354..1375
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1494..1511)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1665..2253)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1745..1764)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2424..3215,3216..3284))
                     /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     3047..3066
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3285..3389)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3457..3475
                     /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 ccagtgaatt cctccaagaa ctcgtacaaa
      421 aatgctctga agtaggtctc gagatcaata ctgggaagac gaaagtcttg cgaaaccgat
      481 tcgctgaccc cagtaaagtc tacttcggta gcccttcccc caccacccag ctcgacgacg
      541 tcgacgagta catctacctc ggtcgtcaaa tcaacgccca aaacaacttg atgccggaaa
      601 tccaccgaag acgtcgagca gcctgggctg cattcaatgg aatcaagaat gccaccgact
      661 ccatcaccga caagaagatt cgtgcgaatc tgttcgactc aattgtcctt ccagcgctca
      721 cctacgtttc agaagcctgg acattcacca aagctctatc cgaacgagta cgaatcacac
      781 atgcctcccc cacaaattat tgatctacat ccccatttgg tatagtgtct tacaggactc
      841 tcaatatgtc agctgcatga taaatggtcc ctatataatt gaattgcaaa tctaaatgtt
      901 tggctaccat aggcaccacg aggttttaga gctagaaata gcaagttaaa ataaggctag
      961 tccgttatca acttgaaaaa gtggcaccga gtcggtgctt tttttcaata atattctaga
     1021 tatccctttt tgaaactaaa atgaaatact ctaaattttt gagttcaatg tttcaactaa
     1081 tcttggttaa aggtaaattc ccggagggac gattttaaag ttgccagagt tgattaaatt
     1141 ggtccaaaaa attttattag gaaactcgaa aaaaaattga ttttttttga caatattcca
     1201 aagaaaatta caaaatctcg acgccaaaca tagtcggctg tgaagcttgg cgtaatcatg
     1261 gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc
     1321 cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc
     1381 gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat
     1441 cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcatatgcgc ttcctcgctc
     1501 actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg
     1561 gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc
     1621 cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc
     1681 ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga
     1741 ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc
     1801 ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat
     1861 agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg
     1921 cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc
     1981 aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga
     2041 gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact
     2101 agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt
     2161 ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag
     2221 cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg
     2281 tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa
     2341 aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata
     2401 tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg
     2461 atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata
     2521 cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg
     2581 gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct
     2641 gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt
     2701 tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc
     2761 tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga
     2821 tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt
     2881 aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc
     2941 atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa
     3001 tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca
     3061 catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca
     3121 aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct
     3181 tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc
     3241 gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa
     3301 tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt
     3361 tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc
     3421 taagaaacca ttattatcat gacattaacc tataaaaata ggcgtatcac gaggcccttt
     3481 cgtc
//
