LOCUS       sequence-334724-        4092 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Donor DNA for TKIT knockin of SEP to NR2A in mouse.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4092)
  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 4092)
  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-GluN2A TKIT donor
FEATURES             Location/Qualifiers
     source          1..4092
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(join(397..1188,1189..1257))
                     /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     1020..1039
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1258..1362)
                     /gene="bla"
                     /label=AmpR promoter
     CDS             1717..2427
                     /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(1744..1772)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     2367..2387
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     primer_bind     complement(3559..3576)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(join(3730..4092,1..226))
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3810..3829)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa
       61 ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt
      121 cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt
      181 ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat
      241 cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat
      301 gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc
      361 aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc
      421 acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta
      481 gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga
      541 cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg
      601 cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc
      661 tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat
      721 cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag
      781 gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat
      841 cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa
      901 ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa
      961 gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga
     1021 taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg
     1081 gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc
     1141 acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg
     1201 aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact
     1261 cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat
     1321 atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgactcga
     1381 gaattcacac cgagcaattc tcacaaagga gatggacatg ggtcgaagaa tgtggatggc
     1441 atcaaggatg agtattctaa agagtgacaa aaacagtcag agctggtgtc gggtttcaca
     1501 ttcgactctc agagcgggtg caggggcagt ctatcctgtt tcctgggccc ctcactgacc
     1561 accacctgcc tctcgctctc ctccacctta cagggaccct aagtggcgac catgggcaga
     1621 ctgggctact ggaccttgct ggtattgccg gcccttctgg tctggcacgg tccggcgcag
     1681 aacgcggcgg cggagaaggg tggcggtggc tctagaagta aaggagaaga acttttcact
     1741 ggagttgtcc caattcttgt tgaattagat ggtgatgtta atgggcacaa attttctgtc
     1801 agtggagagg gtgaaggtga tgcaacatac ggaaaactta cccttaaatt tatttgcact
     1861 actggaaaac tacctgttcc ttggccaaca cttgtcacta ctttaactta tggtgttcaa
     1921 tgcttttcaa gatacccaga tcatatgaaa cggcatgact ttttcaagag tgccatgccc
     1981 gaaggttatg tacaggaaag aactatattt ttcaaagatg acgggaacta caagacacgt
     2041 gctgaagtca agtttgaagg tgataccctt gttaatagaa tcgagttaaa aggtattgat
     2101 tttaaagaag atggaaacat tcttggacac aaattggaat acaactataa cgatcaccag
     2161 gtgtacatca tggcagacaa acaaaagaat ggaatcaaag ctaacttcaa aattagacac
     2221 aacattgaag atggaggcgt tcaactagca gaccattatc aacaaaatac tccaattggc
     2281 gatgggcccg tccttttacc agacaaccat tacctgttta caacttctac tctttcgaaa
     2341 gatcccaacg aaaagagaga ccacatggtc cttcttgagt ttgtaacagc tgctgggatt
     2401 acacatggca tggatgaact atacaagctt ggtggcggtg gctcgggcgg aggtgggtcg
     2461 ggtggcggcg gatcaggcgg tggcggatcc gagaagggta ctccagcgct gaacattgcg
     2521 gtgctgctgg gtcacagcca cgacgtgaca gaacgcgaac ttcgaaatct gtggggcccg
     2581 gagcaggcaa ccggcttgcc cctggatgtg aacgtggtgg cgttattgat gaaccgcact
     2641 gaccctaaga gcctcatcac gcatgtgtgc gacctcatgt ccggggcgcg catccatggc
     2701 ttggtgtttg gagatgatac ggaccaagag gctgtggctc agatgctgga ttttatctcc
     2761 tcacagactt tcatccccat cttgggcatt catgggggtg catctatgat catggctgac
     2821 aaggtaaggc caggaggaga tggactgcca gggtcctgga ggggttgttc tacaaatggc
     2881 agccacctcg cgggggtggg gtggggtaca tctcctattc agttgtacac atatactcaa
     2941 ttcgaaagca ggttaagtgg cctctgacca cgtatcagtc caagccgagc gaattcggat
     3001 ccctcgagcg atacagcgag tcgctctttg tggaggtgac gattacctaa caatcggtcg
     3061 attcgtttga tgttatgttt tgttctcgct ttggttggca ggttacggcc aagttcggta
     3121 agagtgagag ttttacagtc aagtaatgcg tggcaagcca acgttaagct gttgagtcgt
     3181 tttaagtgta attcggggca gaattggtaa agagagtcgt gtaaaatatc gagttcgcac
     3241 atcttgttgt ctgattattg atttttcgcg aaaccattta atcatatgac aagatgtgtg
     3301 tccaccttaa cttaatgatt tttaccaaaa tcattagggg attcatcagc gctgagtgtg
     3361 taaattaatt tttatgccgc agcgggccag caattctcgt gaatcatcgc ttaaacggcc
     3421 tgatttctga atagctatgc gtaagcgggt gttgtctaac attctgcgtt cctctttatc
     3481 ctgtctgaac cggctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat
     3541 tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg
     3601 agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc
     3661 aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcatt
     3721 gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag
     3781 tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc
     3841 cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc
     3901 ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt
     3961 cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt
     4021 atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc
     4081 agccactggt aa
//
