LOCUS       sequence-334033-        3294 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express a nicking sgRNA used for installation of the oncogenic S45F 
            or S45del in Ctnnb1 in mouse liver..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3294)
  AUTHORS   Liu P, Liang SQ, Zheng C, Mintzer E, Zhao YG, Ponnienselvan K, Mir 
            A, Sontheimer EJ, Gao G, Flotte TR, Wolfe SA, Xue W
  TITLE     Improved prime editors enable pathogenic allele correction and 
            cancer modelling in adult mice.
  JOURNAL   Nat Commun. 2021 Apr 9;12(1):2121. doi: 10.1038/s41467-021-22295-w.
  PUBMED    33837189
REFERENCE   2  (bases 1 to 3294)
  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: "Nat 
            Commun."; date: "2021-04-9"; volume: "12(1):2121. doi"; pages: " 
            10.1038/s41467-021-22295-w"
COMMENT     Sequence Label: sgCTNNB1 Nicking
FEATURES             Location/Qualifiers
     source          1..3294
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        15..119
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(120..188,189..980)
                     /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     complement(338..357)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1151..1739
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1640..1659
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1893..1910
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    2027..2048
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(2063..2068,2069..2086,2087..2093)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2101..2117
                     /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)."
     primer_bind     2106..2128
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     2125..2141
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2125..2141
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     2160..2180
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        2162..2180
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        2227..2475
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     2227..2247
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     2398..2417
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        2497..2572
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     primer_bind     complement(2600..2616)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2601..2617)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     complement(2641..2660)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2642..2660)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2670..2687)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2670..2686)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2679..2701)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      2828..3283
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2915..2934)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3125..3146
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
ORIGIN
        1 ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt
       61 atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagta
      121 tgagtattca acatttccgt gtcgccctta ttcccttttt tgcggcattt tgccttcctg
      181 tttttgctca cccagaaacg ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac
      241 gagtgggtta catcgaactg gatctcaaca gcggtaagat ccttgagagt tttcgccccg
      301 aagaacgttt tccaatgatg agcactttta aagttctgct atgtggcgcg gtattatccc
      361 gtattgacgc cgggcaagag caactcggtc gccgcataca ctattctcag aatgacttgg
      421 ttgagtactc accagtcaca gaaaagcatc ttacggatgg catgacagta agagaattat
      481 gcagtgctgc cataaccatg agtgataaca ctgcggccaa cttacttctg acaacgatcg
      541 gaggaccgaa ggagctaacc gcttttttgc acaacatggg ggatcatgta actcgccttg
      601 atcgttggga accggagctg aatgaagcca taccaaacga cgagcgtgac accacgatgc
      661 ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg cgaactactt actctagctt
      721 cccggcaaca attaatagac tggatggagg cggataaagt tgcaggacca cttctgcgct
      781 cggcccttcc ggctggctgg tttattgctg ataaatctgg agccggtgag cgtgggtctc
      841 gcggtatcat tgcagcactg gggccagatg gtaagccctc ccgtatcgta gttatctaca
      901 cgacggggag tcaggcaact atggatgaac gaaatagaca gatcgctgag ataggtgcct
      961 cactgattaa gcattggtaa ctgtcagacc aagtttactc atatatactt tagattgatt
     1021 taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga
     1081 ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca
     1141 aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac
     1201 caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg
     1261 taactggctt cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag
     1321 gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac
     1381 cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt
     1441 taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg
     1501 agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc
     1561 ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc
     1621 gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc
     1681 acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa
     1741 acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt
     1801 tctttcctgc gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg
     1861 ataccgctcg ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag
     1921 agcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc
     1981 acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc
     2041 tcactcatta ggcaccccag gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa
     2101 ttgtgagcgg ataacaattt cacacaggaa acagctatga ccatgattac gccaagcgcg
     2161 caattaaccc tcactaaagg gaacaaaagc tggagctcca ccgcggtggc ggccgccccc
     2221 ttcaccgagg gcctatttcc catgattcct tcatatttgc atatacgata caaggctgtt
     2281 agagagataa ttggaattaa tttgactgta aacacaaaga tattagtaca aaatacgtga
     2341 cgtagaaagt aataatttct tgggtagttt gcagttttaa aattatgttt taaaatggac
     2401 tatcatatgc ttaccgtaac ttgaaagtat ttcgatttct tggctttata tatcttgtgg
     2461 aaaggacgaa acaccggaaa agctgctgtc agccacgttt tagagctaga aatagcaagt
     2521 taaaataagg ctagtccgtt atcaacttga aaaagtggca ccgagtcggt gctttttttg
     2581 aattcgatat caagcttatc gataccgtcg acctcgaggg ggggcccggt acccaattcg
     2641 ccctatagtg agtcgtatta cgcgcgctca ctggccgtcg ttttacaacg tcgtgactgg
     2701 gaaaaccctg gcgttaccca acttaatcgc cttgcagcac atcccccttt cgccagctgg
     2761 cgtaatagcg aagaggcccg caccgatcgc ccttcccaac agttgcgcag cctgaatggc
     2821 gaatgggacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc
     2881 gtgaccgcta cacttgccag cgccctagcg cccgctcctt tcgctttctt cccttccttt
     2941 ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc gggggctccc tttagggttc
     3001 cgatttagtg ctttacggca cctcgacccc aaaaaacttg attagggtga tggttcacgt
     3061 agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc cacgttcttt
     3121 aatagtggac tcttgttcca aactggaaca acactcaacc ctatctcggt ctattctttt
     3181 gatttataag ggattttgcc gatttcggcc tattggttaa aaaatgagct gatttaacaa
     3241 aaatttaacg cgaattttaa caaaatatta acgcttacaa tttaggtggc actt
//
