LOCUS       sequence-332832-        3296 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  SauCas9KKH nicking sgRNA for CCR5.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3296)
  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 3296)
  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: SauCas9KKH CCR5-nicking sgRNA
FEATURES             Location/Qualifiers
     source          1..3296
                     /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(376..392)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(377..393)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     complement(417..436)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(418..436)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(446..463)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(446..462)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(455..477)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      604..1059
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(691..710)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     901..922
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        1085..1189
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1190..1258,1259..2050)
                     /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(1408..1427)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2221..2809
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2710..2729
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2963..2980
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    3097..3118
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(3133..3138,3139..3156,3157..3163)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3171..3187
                     /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     3176..3198
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     3195..3211
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3195..3211
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     3230..3250
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        3232..3250
                     /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 catctttacc agatctcaaa aagttttagt actctggaaa cagaatctac
      301 taaaacaagg caaaatgccg tgtttatctc gtcaacttgt tggcgagatt tttttgaatt
      361 cgatatcaag cttatcgata ccgtcgacct cgaggggggg cccggtaccc aattcgccct
      421 atagtgagtc gtattacgcg cgctcactgg ccgtcgtttt acaacgtcgt gactgggaaa
      481 accctggcgt tacccaactt aatcgccttg cagcacatcc ccctttcgcc agctggcgta
      541 atagcgaaga ggcccgcacc gatcgccctt cccaacagtt gcgcagcctg aatggcgaat
      601 gggacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga
      661 ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct tcctttctcg
      721 ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat
      781 ttagtgcttt acggcacctc gaccccaaaa aacttgatta gggtgatggt tcacgtagtg
      841 ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata
      901 gtggactctt gttccaaact ggaacaacac tcaaccctat ctcggtctat tcttttgatt
      961 tataagggat tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat
     1021 ttaacgcgaa ttttaacaaa atattaacgc ttacaattta ggtggcactt ttcggggaaa
     1081 tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat
     1141 gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagta tgagtattca
     1201 acatttccgt gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca
     1261 cccagaaacg ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac gagtgggtta
     1321 catcgaactg gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgttt
     1381 tccaatgatg agcactttta aagttctgct atgtggcgcg gtattatccc gtattgacgc
     1441 cgggcaagag caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtactc
     1501 accagtcaca gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc
     1561 cataaccatg agtgataaca ctgcggccaa cttacttctg acaacgatcg gaggaccgaa
     1621 ggagctaacc gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga
     1681 accggagctg aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgtagcaat
     1741 ggcaacaacg ttgcgcaaac tattaactgg cgaactactt actctagctt cccggcaaca
     1801 attaatagac tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc
     1861 ggctggctgg tttattgctg ataaatctgg agccggtgag cgtgggtctc gcggtatcat
     1921 tgcagcactg gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag
     1981 tcaggcaact atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa
     2041 gcattggtaa ctgtcagacc aagtttactc atatatactt tagattgatt taaaacttca
     2101 tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc
     2161 ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc
     2221 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
     2281 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
     2341 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
     2401 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
     2461 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
     2521 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
     2581 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
     2641 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
     2701 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
     2761 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
     2821 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc
     2881 gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg
     2941 ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcccaat
     3001 acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc acgacaggtt
     3061 tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc tcactcatta
     3121 ggcaccccag gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa ttgtgagcgg
     3181 ataacaattt cacacaggaa acagctatga ccatgattac gccaagcgcg caattaaccc
     3241 tcactaaagg gaacaaaagc tggagctcca ccgcggtggc ggccgccccc ttcacc
//
