LOCUS       sequence-332902-        3339 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  SauCas9 pegRNA for EMX1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3339)
  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 3339)
  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: SauCas9 EMX1-pegRNA
FEATURES             Location/Qualifiers
     source          1..3339
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..249
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     1..21
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     172..191
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        273..348
                     /label=Sa gRNA scaffold
                     /note="guide RNA scaffold for the Staphylococcus aureus 
                     CRISPR/Cas9 system"
     primer_bind     complement(419..435)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(420..436)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     complement(460..479)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(461..479)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(489..506)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(489..505)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(498..520)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      647..1102
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(734..753)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     944..965
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        1128..1232
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1233..1301,1302..2093)
                     /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(1451..1470)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2264..2852
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2753..2772
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3006..3023
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    3140..3161
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(3176..3181,3182..3199,3200..3206)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3214..3230
                     /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     3219..3241
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     3238..3254
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3238..3254
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     3273..3293
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        3275..3293
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
ORIGIN
        1 gagggcctat ttcccatgat tccttcatat ttgcatatac gatacaaggc tgttagagag
       61 ataattggaa ttaatttgac tgtaaacaca aagatattag tacaaaatac gtgacgtaga
      121 aagtaataat ttcttgggta gtttgcagtt ttaaaattat gttttaaaat ggactatcat
      181 atgcttaccg taacttgaaa gtatttcgat ttcttggctt tatatatctt gtggaaagga
      241 cgaaacaccg cagaagctgg aggaggaagg gcgttttagt actctggaaa cagaatctac
      301 taaaacaagg caaaatgccg tgtttatctc gtcaacttgt tggcgagatt cttctgctcg
      361 gactcaggcc aagcttcttc ctcctccagc ttttttttga attcgatatc aagcttatcg
      421 ataccgtcga cctcgagggg gggcccggta cccaattcgc cctatagtga gtcgtattac
      481 gcgcgctcac tggccgtcgt tttacaacgt cgtgactggg aaaaccctgg cgttacccaa
      541 cttaatcgcc ttgcagcaca tccccctttc gccagctggc gtaatagcga agaggcccgc
      601 accgatcgcc cttcccaaca gttgcgcagc ctgaatggcg aatgggacgc gccctgtagc
      661 ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgccagc
      721 gccctagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt
      781 ccccgtcaag ctctaaatcg ggggctccct ttagggttcc gatttagtgc tttacggcac
      841 ctcgacccca aaaaacttga ttagggtgat ggttcacgta gtgggccatc gccctgatag
      901 acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact cttgttccaa
      961 actggaacaa cactcaaccc tatctcggtc tattcttttg atttataagg gattttgccg
     1021 atttcggcct attggttaaa aaatgagctg atttaacaaa aatttaacgc gaattttaac
     1081 aaaatattaa cgcttacaat ttaggtggca cttttcgggg aaatgtgcgc ggaaccccta
     1141 tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat
     1201 aaatgcttca ataatattga aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc
     1261 ttattccctt ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctggtga
     1321 aagtaaaaga tgctgaagat cagttgggtg cacgagtggg ttacatcgaa ctggatctca
     1381 acagcggtaa gatccttgag agttttcgcc ccgaagaacg ttttccaatg atgagcactt
     1441 ttaaagttct gctatgtggc gcggtattat cccgtattga cgccgggcaa gagcaactcg
     1501 gtcgccgcat acactattct cagaatgact tggttgagta ctcaccagtc acagaaaagc
     1561 atcttacgga tggcatgaca gtaagagaat tatgcagtgc tgccataacc atgagtgata
     1621 acactgcggc caacttactt ctgacaacga tcggaggacc gaaggagcta accgcttttt
     1681 tgcacaacat gggggatcat gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag
     1741 ccataccaaa cgacgagcgt gacaccacga tgcctgtagc aatggcaaca acgttgcgca
     1801 aactattaac tggcgaacta cttactctag cttcccggca acaattaata gactggatgg
     1861 aggcggataa agttgcagga ccacttctgc gctcggccct tccggctggc tggtttattg
     1921 ctgataaatc tggagccggt gagcgtgggt ctcgcggtat cattgcagca ctggggccag
     1981 atggtaagcc ctcccgtatc gtagttatct acacgacggg gagtcaggca actatggatg
     2041 aacgaaatag acagatcgct gagataggtg cctcactgat taagcattgg taactgtcag
     2101 accaagttta ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga
     2161 tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt
     2221 tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc
     2281 tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc
     2341 cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac
     2401 caaatactgt tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac
     2461 cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt
     2521 cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct
     2581 gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat
     2641 acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt
     2701 atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg
     2761 cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt
     2821 gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt
     2881 tcctggcctt ttgctggcct tttgctcaca tgttctttcc tgcgttatcc cctgattctg
     2941 tggataaccg tattaccgcc tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg
     3001 agcgcagcga gtcagtgagc gaggaagcgg aagagcgccc aatacgcaaa ccgcctctcc
     3061 ccgcgcgttg gccgattcat taatgcagct ggcacgacag gtttcccgac tggaaagcgg
     3121 gcagtgagcg caacgcaatt aatgtgagtt agctcactca ttaggcaccc caggctttac
     3181 actttatgct tccggctcgt atgttgtgtg gaattgtgag cggataacaa tttcacacag
     3241 gaaacagcta tgaccatgat tacgccaagc gcgcaattaa ccctcactaa agggaacaaa
     3301 agctggagct ccaccgcggt ggcggccgcc cccttcacc
//
