LOCUS       sequence-496311-        3900 bp    DNA     circular SYN 07-JAN-2026
DEFINITION  Expresses nucleus-localized Slug-nCas9 (D10A) fused to PolI5M-Delta 
            and mCherry using a CMV promoter. Includes a GFP cassette flanked by
            BsmbI cut sites to knock in and coexpress user-defined gRNAs..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3900)
  AUTHORS   Hurtado JE, Schieferecke AJ, Halperin SO, Guan J, Aidlen D, Schaffer
            DV, Dueber JE
  TITLE     Nickase fidelity drives EvolvR-mediated diversification in mammalian
            cells.
  JOURNAL   Nat Commun. 2025 Apr 19;16(1):3723. doi: 10.1038/s41467-025-58414-0.
  PUBMED    40253348
REFERENCE   2  (bases 1 to 3900)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Jan 7, 2026 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nat 
            Commun."; date: "2025-04-19"; volume: "16(1):3723. doi"; pages: " 
            10.1038/s41467-025-58414-0"
COMMENT     Sequence Label: pEvolvR-mCherry-Slug-nCas9(D10A)-PolI5M-Delta
FEATURES             Location/Qualifiers
     source          1..3900
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      1..589
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     490..509
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2833..3624,3625..3693))
                     /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     3456..3475
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3694..3798)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
       61 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
      121 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
      181 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
      241 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
      301 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
      361 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
      421 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
      481 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
      541 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
      601 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc
      661 gttatcccct gattctgtgg ataaccgtgc ggccgcccct gaattcgcat ctagaataaa
      721 tcgcagccaa gtgagtgaat agatgacgca ccacggtcag acacggccac atcgtatctc
      781 acaggagcaa gcgcgatagg agcactcaca catagtacgg tgatccgctg actcctttgc
      841 ccaaataaga cgtgagcccg tctcagatgc acacactggc ttaagatgac aacgtgcaga
      901 aaatatgctc ttggcaatta cgtagttgta gaagctgggg atggcttgca aagaggccct
      961 tgggcgttga tgggtaaggg cggatctcct ggggcagcgg atggacagaa gccagttgta
     1021 gattctggct ccgcagtatc ccgagtttca gccgtccctg cctgggtcct gcgaccttcc
     1081 cctggattgc aaaaggtaaa caatagcata cagaaagaaa cgatagcgta gaaaaggcgg
     1141 gtaagataca cttaataaaa cgtctacgcg aaagcagcat cccagataac cgggtaagat
     1201 ggaggtgctc acggctagat tgagtgcgct gcggtttgct cggcacggtc gagccagtat
     1261 cggcaatgtc ccggtatcca attttcacgc aaaatatgtc ttcccggata ttccgttgct
     1321 gaaaaacaat agaaatcggt tttctcaacg atatcgacca gtgaaaatac agctaaataa
     1381 ctcgttggtc ggaaacttaa ttgtggtttg tgtttctttt ttaccctttt ggtaccgatt
     1441 gttgtttact caatttggac aaacaaagtc gctcgtttca aggtgcataa tttgcatcca
     1501 cctcttaaaa atgcagtatc tttccttcgt ttttctccat ccaactttat attctttttt
     1561 tctttcagtt caattgctag aatagagact catcagatct atggagaacc gtgagccacc
     1621 actgttgcca gcccgttgga gcagcgccta tgtctcttat tggagcccga tgctgccgga
     1681 tgaccagctg accagcggct attgctggtt cgactatgaa cgtgacatct gtcgtattga
     1741 cggcctgttc aatccgtgga gcgagcgtga tactggttat cgcctgtgga tgtcggaggt
     1801 tggtaatgcg gccagcggcc gtacctggaa acaaaaagtc gcctatggtc gtgagcgtac
     1861 cgccctgggt gaacagctgt gtgagcgtcc gctggatgat gagactggcc cttttgccga
     1921 attgttcctg ccacgcgatg tcctgcgccg tctgggtgcc cgtcacattg gccgtcgcgt
     1981 ggttctgggt cgcgaagcgg acggttggcg ttaccagcgc ccaggtaaag gtccgagcac
     2041 cctgtacctg gatgcggcga gcggcactcc actgcgcatg gtcaccggcg atgaagcgtc
     2101 gcgtgcaagc ctgcgtgatt ttccgaatgt gagcgaggcg gagatcccgg acgcggtttt
     2161 cgcggccaag cgcggatcct ttccaccaga agttgaagaa caagatgatg gtactttacc
     2221 aatgtcttgt gctcaagaat ctggtatgga tagacatggt tcattgggaa gaggtagaag
     2281 atccaaattg taataatggc taattacaca atggcttggt ttttttctat aagcccgtcc
     2341 cagtttgtag accagattaa ataaacttcg gctgcagaca ggaaaagctc tccgagtaga
     2401 aagccgaccc acaagccagc aactcctaac ccaaagtaga agccaaacaa caactccaga
     2461 ggaatggcaa tcatgtagta ggccgtcaga ttcaacactg agccgatgtt ttggcggccc
     2521 tggcctctca aaagggagga aaaaatggcg ttggctggtt ctgagacggc agacctagac
     2581 cacatgaggc tgatgtagga cagccaccag tggcagctaa tcaacaccgc aagatgccga
     2641 tgcacgctca tatcatcgtc gtcagcctgg tagccattcg acatacggat cagggaactc
     2701 gaacccagta cactagtgca ctgcagtaca gcggccgcga ttatcaaaaa ggatcttcac
     2761 ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac
     2821 ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt
     2881 tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt
     2941 accatctggc cccagtgctg caatgatacc gcgggaccca cgctcaccgg ctccagattt
     3001 atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc
     3061 cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa
     3121 tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg
     3181 tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt
     3241 gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc
     3301 agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt
     3361 aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg
     3421 gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac
     3481 tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc
     3541 gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt
     3601 tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg
     3661 aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat attattgaag
     3721 catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa
     3781 acaaataggg gttccgcgca catttccccg aaaagtgcca cctgtcatga ccaaaatccc
     3841 ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc
//
