LOCUS       sequence-334036-        3320 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pegRNA used for installation (via TCC deletion) of the oncogenic 
            S45del in Ctnnb1 in mouse liver..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3320)
  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 3320)
  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: pegCTNNB1 S45del
FEATURES             Location/Qualifiers
     source          1..3320
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        5..109
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(110..178,179..970)
                     /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(328..347)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1141..1729
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1630..1649
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1883..1900
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    2017..2038
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(2053..2058,2059..2076,2077..2083)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2091..2107
                     /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     2096..2118
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     2115..2131
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2115..2131
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     2150..2170
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        2152..2170
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        2217..2465
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     2217..2237
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     2388..2407
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        2487..2562
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     primer_bind     complement(2616..2632)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2617..2633)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     complement(2657..2676)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2658..2676)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2686..2703)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2686..2702)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2695..2717)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      2844..3299
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2931..2950)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3141..3162
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
ORIGIN
        1 tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat
       61 gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagta tgagtattca
      121 acatttccgt gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca
      181 cccagaaacg ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac gagtgggtta
      241 catcgaactg gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgttt
      301 tccaatgatg agcactttta aagttctgct atgtggcgcg gtattatccc gtattgacgc
      361 cgggcaagag caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtactc
      421 accagtcaca gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc
      481 cataaccatg agtgataaca ctgcggccaa cttacttctg acaacgatcg gaggaccgaa
      541 ggagctaacc gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga
      601 accggagctg aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgtagcaat
      661 ggcaacaacg ttgcgcaaac tattaactgg cgaactactt actctagctt cccggcaaca
      721 attaatagac tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc
      781 ggctggctgg tttattgctg ataaatctgg agccggtgag cgtgggtctc gcggtatcat
      841 tgcagcactg gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag
      901 tcaggcaact atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa
      961 gcattggtaa ctgtcagacc aagtttactc atatatactt tagattgatt taaaacttca
     1021 tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc
     1081 ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc
     1141 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
     1201 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
     1261 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
     1321 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
     1381 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
     1441 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
     1501 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
     1561 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
     1621 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
     1681 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
     1741 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc
     1801 gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg
     1861 ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcccaat
     1921 acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc acgacaggtt
     1981 tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc tcactcatta
     2041 ggcaccccag gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa ttgtgagcgg
     2101 ataacaattt cacacaggaa acagctatga ccatgattac gccaagcgcg caattaaccc
     2161 tcactaaagg gaacaaaagc tggagctcca ccgcggtggc ggccgccccc ttcaccgagg
     2221 gcctatttcc catgattcct tcatatttgc atatacgata caaggctgtt agagagataa
     2281 ttggaattaa tttgactgta aacacaaaga tattagtaca aaatacgtga cgtagaaagt
     2341 aataatttct tgggtagttt gcagttttaa aattatgttt taaaatggac tatcatatgc
     2401 ttaccgtaac ttgaaagtat ttcgatttct tggctttata tatcttgtgg aaaggacgaa
     2461 acaccgaggg ttgcccttgc cactcagttt tagagctaga aatagcaagt taaaataagg
     2521 ctagtccgtt atcaacttga aaaagtggga ccgagtcggt ccacagctcc tctgagtggc
     2581 aagggcaatt tttttgaatt cgatatcaag cttatcgata ccgtcgacct cgaggggggg
     2641 cccggtaccc aattcgccct atagtgagtc gtattacgcg cgctcactgg ccgtcgtttt
     2701 acaacgtcgt gactgggaaa accctggcgt tacccaactt aatcgccttg cagcacatcc
     2761 ccctttcgcc agctggcgta atagcgaaga ggcccgcacc gatcgccctt cccaacagtt
     2821 gcgcagcctg aatggcgaat gggacgcgcc ctgtagcggc gcattaagcg cggcgggtgt
     2881 ggtggttacg cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc
     2941 tttcttccct tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg
     3001 gctcccttta gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgatta
     3061 gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt
     3121 ggagtccacg ttctttaata gtggactctt gttccaaact ggaacaacac tcaaccctat
     3181 ctcggtctat tcttttgatt tataagggat tttgccgatt tcggcctatt ggttaaaaaa
     3241 tgagctgatt taacaaaaat ttaacgcgaa ttttaacaaa atattaacgc ttacaattta
     3301 ggtggcactt ttcggggaaa
//
