LOCUS       sequence-435552-        2766 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of 2x-deadEGFP(A111V/L202S) and aptamer-gRNAs 
            to correct both mutations for MOBE1-2.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2766)
  AUTHORS   Cowan QT, Gu S, Gu W, Ranzau BL, Simonson TS, Komor AC
  TITLE     Development of multiplexed orthogonal base editor (MOBE) systems.
  JOURNAL   Nat Biotechnol. 2024 May 21. doi: 10.1038/s41587-024-02240-0.
  PUBMED    38773305
REFERENCE   2  (bases 1 to 2766)
  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 
            Biotechnol. 2024 May 21. doi: 10.1038/s41587-024-02240-0."
COMMENT     Sequence Label: Fluorescent reporter for MOBE1-2
FEATURES             Location/Qualifiers
     source          1..2766
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(68..656)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(148..167)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(827..1618,1619..1687))
                     /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     1450..1469
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1688..1792)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     1860..1878
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(1916..1938)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     2038..2057
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(2074..2529)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2211..2232)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2423..2442
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     repeat_region   2604..2744
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   2615..2744
                     /label=AAV2 ITR
     primer_bind     join(2751..2766,1)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     join(2751..2766,1)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 ctagcgacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc
       61 tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc
      121 agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc
      181 tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt
      241 cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg
      301 ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat
      361 ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag
      421 ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt
      481 ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc
      541 cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta
      601 gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag
      661 atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga
      721 ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa
      781 gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa
      841 tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc
      901 ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga
      961 taccgcggct tccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa
     1021 gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt
     1081 gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg
     1141 ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc
     1201 aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg
     1261 gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag
     1321 cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt
     1381 actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt
     1441 caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac
     1501 gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac
     1561 ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag
     1621 caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa
     1681 tactcatact cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga
     1741 gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc
     1801 cccgaaaagt gccacctgac gtctaagaaa ccattattat catgacatta acctataaaa
     1861 ataggcgtat cacgaggccc tttcgtctcg cgcgtttcgg tgatgacggt gaaaacctct
     1921 gacacatgca gctcccggag acggtcacag cttgtctgta agcggatgcc gggagcagac
     1981 aagcccgtca gggcgcgtca gcgggtgttg gcgggtgtcg gggctggctt aactatgcgg
     2041 catcagagca gattgtactg agagtgcacc ataaaattgt aaacgttaat attttgttaa
     2101 aattcgcgtt aaatttttgt taaatcagct cattttttaa ccaatagacc gaaatcggca
     2161 aaatccctta taaatcaaaa gaatagcccg agatagagtt gagtgttgtt ccagtttgga
     2221 acaagagtcc actattaaag aacgtggact ccaacgtcaa agggcgaaaa accgtctatc
     2281 agggcgatgg cccactacgt gaaccatcac ccaaatcaag ttttttgggg tcgaggtgcc
     2341 gtaaagcact aaatcggaac cctaaaggga gcccccgatt tagagcttga cggggaaagc
     2401 cggcgaacgt ggcgagaaag gaagggaaga aagcgaaagg agcgggcgct aaggcgctgg
     2461 caagtgtagc ggtcacgctg cgcgtaacca ccacacccgc cgcgcttaat gcgccgctac
     2521 agggcgcgta ctatggttgc tttgacgtat gcggtgtgaa ataccgcaca gatgcgtaag
     2581 gagaaaatac cgcatcaggc gcccctgcag gcagctgcgc gctcgctcgc tcactgaggc
     2641 cgcccgggca aagcccgggc gtcgggcgac ctttggtcgc ccggcctcag tgagcgagcg
     2701 agcgcgcaga gagggagtgg ccaactccat cactaggggt tcctgcggcc caggaaacag
     2761 ctatga
//
