LOCUS       sequence-370782-        3484 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  miniMos Arm 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: pMLS719
FEATURES             Location/Qualifiers
     source          1..3484
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     108..127
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     321..343
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     335..352
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     336..352
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        359..858
                     /label=CeU6 promoter
                     /note="promoter for the C. elegans K09B11.12 U6 snRNA gene 
                     (Friedland et al., 2013)"
     misc_RNA        880..955
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     misc_feature    963..1199
                     /label=3' CeU6
                     /note="region downstream of the C. elegans K09B11.12 U6 
                     snRNA gene"
     primer_bind     complement(1218..1234)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1218..1234)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1231..1253)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1242..1258
                     /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(1266..1272,1273..1290,1291..1296))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1311..1332
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1451..1468)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1622..2210)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1702..1721)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2381..3172,3173..3241))
                     /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     3004..3023
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3242..3346)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3414..3432
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(join(3470..3484,1..8))
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
ORIGIN
        1 gctcccggag acggtcacag cttgtctgta agcggatgcc gggagcagac aagcccgtca
       61 gggcgcgtca gcgggtgttg gcgggtgtcg gggctggctt aactatgcgg catcagagca
      121 gattgtactg agagtgcacc atatgcggtg tgaaataccg cacagatgcg taaggagaaa
      181 ataccgcatc aggcgccatt cgccattcag gctgcgcaac tgttgggaag ggcgatcggt
      241 gcgggcctct tcgctattac gccagctggc gaaaggggga tgtgctgcaa ggcgattaag
      301 ttgggtaacg ccagggtttt cccagtcacg acgttgtaaa acgacggcca gtgaattcct
      361 ccaagaactc gtacaaaaat gctctgaagt aggtctcgag atcaatactg ggaagacgaa
      421 agtcttgcga aaccgattcg ctgaccccag taaagtctac ttcggtagcc cttcccccac
      481 cacccagctc gacgacgtcg acgagtacat ctacctcggt cgtcaaatca acgcccaaaa
      541 caacttgatg ccggaaatcc accgaagacg tcgagcagcc tgggctgcat tcaatggaat
      601 caagaatgcc accgactcca tcaccgacaa gaagattcgt gcgaatctgt tcgactcaat
      661 tgtccttcca gcgctcacct acgtttcaga agcctggaca ttcaccaaag ctctatccga
      721 acgagtacga atcacacatg cctcccccac aaattattga tctacatccc catttggtat
      781 agtgtcttac aggactctca atatgtcagc tgcatgataa atggtcccta tataattgaa
      841 ttgcaaatct aaatgtttga ctgtcgaacc accatagttg ttttagagct agaaatagca
      901 agttaaaata aggctagtcc gttatcaact tgaaaaagtg gcaccgagtc ggtgcttttt
      961 ttcaataata ttctagatat ccctttttga aactaaaatg aaatactcta aatttttgag
     1021 ttcaatgttt caactaatct tggttaaagg taaattcccg gagggacgat tttaaagttg
     1081 ccagagttga ttaaattggt ccaaaaaatt ttattaggaa actcgaaaaa aaattgattt
     1141 tttttgacaa tattccaaag aaaattacaa aatctcgacg ccaaacatag tcggctgtga
     1201 agcttggcgt aatcatggtc atagctgttt cctgtgtgaa attgttatcc gctcacaatt
     1261 ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta atgagtgagc
     1321 taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc
     1381 cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctca
     1441 tatgcgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat
     1501 cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga
     1561 acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt
     1621 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt
     1681 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc
     1741 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa
     1801 gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct
     1861 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta
     1921 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg
     1981 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc
     2041 ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta
     2101 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg
     2161 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt
     2221 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg
     2281 tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta
     2341 aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg
     2401 aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg
     2461 tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc
     2521 gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg
     2581 agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg
     2641 aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag
     2701 gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat
     2761 caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc
     2821 cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc
     2881 ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa
     2941 ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac
     3001 gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt
     3061 cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc
     3121 gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa
     3181 caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca
     3241 tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat
     3301 acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa
     3361 aagtgccacc tgacgtctaa gaaaccatta ttatcatgac attaacctat aaaaataggc
     3421 gtatcacgag gccctttcgt ctcgcgcgtt tcggtgatga cggtgaaaac ctctgacaca
     3481 tgca
//
