LOCUS       sequence-334643-        3924 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Donor DNA for TKIT knockin of SEP to Gabrb2 in mouse.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3924)
  AUTHORS   Fang H, Bygrave AM, Roth RH, Johnson RC, Huganir RL
  TITLE     An optimized CRISPR/Cas9 approach for precise genome editing in 
            neurons.
  JOURNAL   Elife. 2021 Mar 10;10. pii: 65202. doi: 10.7554/eLife.65202.
  PUBMED    33689678
REFERENCE   2  (bases 1 to 3924)
  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: "Elife."; 
            date: "2021-03-10"; volume: "10. pii: 65202. doi"; pages: " 
            10.7554/eLife.65202"
COMMENT     Sequence Label: pMiniT:SEP-beta2 TKIT donor
FEATURES             Location/Qualifiers
     source          1..3924
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(87..104)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(258..846)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(338..357)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1017..1808,1809..1877))
                     /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     1640..1659
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1878..1982)
                     /gene="bla"
                     /label=AmpR promoter
     CDS             2354..3064
                     /codon_start=1
                     /product="pH-sensitive mutant of green fluorescent protein 
                     (Ng et al., 2002)"
                     /label=superecliptic pHluorin
                     /note="excited strongly by blue light at neutral pH but 
                     minimally fluorescent at acidic pH"
                     /translation="SKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLKF
                     ICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTIFFKDDGN
                     YKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNDHQVYIMADKQKNGIKAN
                     FKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLFTTSTLSKDPNEKRDHMVLLEF
                     VTAAGITHGMDELYK"
     primer_bind     complement(2381..2409)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     3004..3024
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
ORIGIN
        1 tctttatcct gtctgaaccg gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt
       61 ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg
      121 ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg
      181 gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag
      241 gccgcattgc tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga
      301 cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct
      361 ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc
      421 tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg
      481 gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc
      541 tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca
      601 ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag
      661 ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct
      721 ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc
      781 accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga
      841 tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca
      901 cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat
      961 taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac
     1021 caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt
     1081 gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt
     1141 gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag
     1201 ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct
     1261 attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt
     1321 gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc
     1381 tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt
     1441 agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg
     1501 gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg
     1561 actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct
     1621 tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc
     1681 attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt
     1741 tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt
     1801 tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg
     1861 aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta tcagggttat
     1921 tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg
     1981 cgcacatttc cccgactcga gaattcagtc tggtcacact ctaagggggg tgcgcatgcg
     2041 ctccacatta gaattactgc actgggcaga ctaagttgga tctcctctct tcagtgaatc
     2101 cctcaatccc accagaacct aaagggatgt ggagagtccg gaaaaggggc tactttggga
     2161 tttggtcatt tcccttaata atcgccgctg tctgtgctca gaggtaagtg tccctttcct
     2221 ttctcacttg tcagggctct gtctgcatac ctctctgtga gcagggctgt tgaagatgcc
     2281 gtgctgtctt ctttattcat tcctggttat tgtcattttt ttctttagtg tcaatggtgg
     2341 cggtggctct agaagtaaag gagaagaact tttcactgga gttgtcccaa ttcttgttga
     2401 attagatggt gatgttaatg ggcacaaatt ttctgtcagt ggagagggtg aaggtgatgc
     2461 aacatacgga aaacttaccc ttaaatttat ttgcactact ggaaaactac ctgttccttg
     2521 gccaacactt gtcactactt taacttatgg tgttcaatgc ttttcaagat acccagatca
     2581 tatgaaacgg catgactttt tcaagagtgc catgcccgaa ggttatgtac aggaaagaac
     2641 tatatttttc aaagatgacg ggaactacaa gacacgtgct gaagtcaagt ttgaaggtga
     2701 tacccttgtt aatagaatcg agttaaaagg tattgatttt aaagaagatg gaaacattct
     2761 tggacacaaa ttggaataca actataacga tcaccaggtg tacatcatgg cagacaaaca
     2821 aaagaatgga atcaaagcta acttcaaaat tagacacaac attgaagatg gaggcgttca
     2881 actagcagac cattatcaac aaaatactcc aattggcgat gggcccgtcc ttttaccaga
     2941 caaccattac ctgtttacaa cttctactct ttcgaaagat cccaacgaaa agagagacca
     3001 catggtcctt cttgagtttg taacagctgc tgggattaca catggcatgg atgaactata
     3061 caagcttggt ggcggtggct cgggcggagg tgggtcgggt ggcggcggat caggcggtgg
     3121 cggatccagt gtcaatgacc ctagtaatat gtcgctggtt aaagagacgg tggacagact
     3181 gttgaaaggc tatgacattc gtctgagacc agattttgga ggtaatggga ttgccttgca
     3241 aataagaaac agcccagctc cagcccaagc tgtcttttgg gctttgtgtg cactctcttt
     3301 cctctcctta gttgtgggcg catgtgatgt atagaatgtt ttcctatttt accttgctga
     3361 gaaacagatc caaacttaag ccagtttctg tccagcactt gagaggtctt tcaagaagag
     3421 gcgtaccaaa acatcaaagg gggaattcgg atccctcgag cgatacagcg agtcgctctt
     3481 tgtggaggtg acgattacct aacaatcggt cgattcgttt gatgttatgt tttgttctcg
     3541 ctttggttgg caggttacgg ccaagttcgg taagagtgag agttttacag tcaagtaatg
     3601 cgtggcaagc caacgttaag ctgttgagtc gttttaagtg taattcgggg cagaattggt
     3661 aaagagagtc gtgtaaaata tcgagttcgc acatcttgtt gtctgattat tgatttttcg
     3721 cgaaaccatt taatcatatg acaagatgtg tgtccacctt aacttaatga tttttaccaa
     3781 aatcattagg ggattcatca gcgctgagtg tgtaaattaa tttttatgcc gcagcgggcc
     3841 agcaattctc gtgaatcatc gcttaaacgg cctgatttct gaatagctat gcgtaagcgg
     3901 gtgttgtcta acattctgcg ttcc
//
