LOCUS       sequence-332901-        3331 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  SpyCas9-pegRNA for CCR5.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3331)
  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 3331)
  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: Spy CCR5-pegRNA
FEATURES             Location/Qualifiers
     source          1..3331
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    121..142
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(157..162,163..180,181..187)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    195..211
                     /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     200..222
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     219..235
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     219..235
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     254..274
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        256..274
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        321..569
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     321..341
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     492..511
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     misc_RNA        591..666
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     primer_bind     complement(731..747)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(732..748)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     complement(772..791)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(773..791)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(801..818)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(801..817)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(810..832)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      959..1414
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1046..1065)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1256..1277
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        1440..1544
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1545..1613,1614..2405)
                     /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(1763..1782)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2576..3164
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3065..3084
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     join(3318..3331,1..4)
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 cgaggaagcg gaagagcgcc caatacgcaa accgcctctc cccgcgcgtt ggccgattca
       61 ttaatgcagc tggcacgaca ggtttcccga ctggaaagcg ggcagtgagc gcaacgcaat
      121 taatgtgagt tagctcactc attaggcacc ccaggcttta cactttatgc ttccggctcg
      181 tatgttgtgt ggaattgtga gcggataaca atttcacaca ggaaacagct atgaccatga
      241 ttacgccaag cgcgcaatta accctcacta aagggaacaa aagctggagc tccaccgcgg
      301 tggcggccgc ccccttcacc gagggcctat ttcccatgat tccttcatat ttgcatatac
      361 gatacaaggc tgttagagag ataattggaa ttaatttgac tgtaaacaca aagatattag
      421 tacaaaatac gtgacgtaga aagtaataat ttcttgggta gtttgcagtt ttaaaattat
      481 gttttaaaat ggactatcat atgcttaccg taacttgaaa gtatttcgat ttcttggctt
      541 tatatatctt gtggaaagga cgaaacaccg agatgactat ctttaatgtc gttttagagc
      601 tagaaatagc aagttaaaat aaggctagtc cgttatcaac ttgaaaaagt gggaccgagt
      661 cggtccttac acctgcagct ctcattttcc ttatattaaa gatagtcatc ttttttgata
      721 tcaagcttat cgataccgtc gacctcgagg gggggcccgg tacccaattc gccctatagt
      781 gagtcgtatt acgcgcgctc actggccgtc gttttacaac gtcgtgactg ggaaaaccct
      841 ggcgttaccc aacttaatcg ccttgcagca catccccctt tcgccagctg gcgtaatagc
      901 gaagaggccc gcaccgatcg cccttcccaa cagttgcgca gcctgaatgg cgaatgggac
      961 gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct
     1021 acacttgcca gcgccctagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg
     1081 ttcgccggct ttccccgtca agctctaaat cgggggctcc ctttagggtt ccgatttagt
     1141 gctttacggc acctcgaccc caaaaaactt gattagggtg atggttcacg tagtgggcca
     1201 tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga
     1261 ctcttgttcc aaactggaac aacactcaac cctatctcgg tctattcttt tgatttataa
     1321 gggattttgc cgatttcggc ctattggtta aaaaatgagc tgatttaaca aaaatttaac
     1381 gcgaatttta acaaaatatt aacgcttaca atttaggtgg cacttttcgg ggaaatgtgc
     1441 gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac
     1501 aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtatgagt attcaacatt
     1561 tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt gctcacccag
     1621 aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg ggttacatcg
     1681 aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa cgttttccaa
     1741 tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc
     1801 aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag tactcaccag
     1861 tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt gctgccataa
     1921 ccatgagtga taacactgcg gccaacttac ttctgacaac gatcggagga ccgaaggagc
     1981 taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt tgggaaccgg
     2041 agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgta gcaatggcaa
     2101 caacgttgcg caaactatta actggcgaac tacttactct agcttcccgg caacaattaa
     2161 tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc cttccggctg
     2221 gctggtttat tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt atcattgcag
     2281 cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg gggagtcagg
     2341 caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg attaagcatt
     2401 ggtaactgtc agaccaagtt tactcatata tactttagat tgatttaaaa cttcattttt
     2461 aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac
     2521 gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag
     2581 atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg
     2641 tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca
     2701 gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac cacttcaaga
     2761 actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca
     2821 gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc
     2881 agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca
     2941 ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa
     3001 aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc
     3061 cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc
     3121 gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg
     3181 cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt cctgcgttat
     3241 cccctgattc tgtggataac cgtattaccg cctttgagtg agctgatacc gctcgccgca
     3301 gccgaacgac cgagcgcagc gagtcagtga g
//
