LOCUS       sequence-334638-        3815 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Donor DNA for TKIT knockin of SEP to GluA2 in mouse.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3815)
  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 3815)
  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-GluA2 TKIT donor
FEATURES             Location/Qualifiers
     source          1..3815
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(168..756)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(248..267)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(927..1718,1719..1787))
                     /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     1550..1569
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1788..1892)
                     /gene="bla"
                     /label=AmpR promoter
     CDS             2225..2935
                     /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(2252..2280)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     2875..2895
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     primer_bind     complement(join(3812..3815,1..14))
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag
       61 gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa
      121 ggccagcaaa aggccaggaa ccgtaaaaag gccgcattgc tggcgttttt ccataggctc
      181 cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca
      241 ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg
      301 accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct
      361 catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt
      421 gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag
      481 tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc
      541 agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac
      601 actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga
      661 gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc
      721 aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg
      781 gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca
      841 aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt
      901 atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca
      961 gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg
     1021 atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca
     1081 ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt
     1141 cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt
     1201 agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca
     1261 cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca
     1321 tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga
     1381 agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact
     1441 gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga
     1501 gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg
     1561 ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc
     1621 tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga
     1681 tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat
     1741 gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt
     1801 caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt
     1861 atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgactcga gaattctgac
     1921 gtcattatta tcagggcctc tattgtgtca aaaggcatac ctgggagata aggattcttc
     1981 tgccttcact tcgtggtttt agcagctcct tgctaaattt cgacctcaaa atgcagagga
     2041 tctaatttgc tgaggaaaac ggtcaaagaa ggaaaaggag gaaagggaaa cgaggggata
     2101 ttttgtggat gctctacttt tcttggaaat gcaaaagatt atgcatattt ctgtcctcct
     2161 ttctcctgtt ttatggggac tgatttttgg tgtctcttct aacactagtg gtggcggtgg
     2221 ctcgagtaaa ggagaagaac ttttcactgg agttgtccca attcttgttg aattagatgg
     2281 tgatgttaat gggcacaaat tttctgtcag tggagagggt gaaggtgatg caacatacgg
     2341 aaaacttacc cttaaattta tttgcactac tggaaaacta cctgttcctt ggccaacact
     2401 tgtcactact ttaacttatg gtgttcaatg cttttcaaga tacccagatc atatgaaacg
     2461 gcatgacttt ttcaagagtg ccatgcccga aggttatgta caggaaagaa ctatattttt
     2521 caaagatgac gggaactaca agacacgtgc tgaagtcaag tttgaaggtg atacccttgt
     2581 taatagaatc gagttaaaag gtattgattt taaagaagat ggaaacattc ttggacacaa
     2641 attggaatac aactataacg atcaccaggt gtacatcatg gcagacaaac aaaagaatgg
     2701 aatcaaagct aacttcaaaa ttagacacaa cattgaagat ggaggcgttc aactagcaga
     2761 ccattatcaa caaaatactc caattggcga tgggcccgtc cttttaccag acaaccatta
     2821 cctgtttaca acttctactc tttcgaaaga tcccaacgaa aagagagacc acatggtcct
     2881 tcttgagttt gtaacagctg ctgggattac acatggcatg gatgaactat acaaaggtgg
     2941 cggtggctcg ggcggaggtg ggtcgggtgg cggcggatca ggcggtggcg gatccagcat
     3001 acagataggt aggtaccttt tgtgttatgc ctttgaattg tgcataattt ggtggtaatc
     3061 tttgttcaca gtgtgcaaat taattcatgt catgtagact tatgtcatac cttgactgca
     3121 ttccagattg aaactaccta gaatatacat gattttctta ggctgaagca aaaagaaatg
     3181 actatgcctt ttgacacaac ccaggtttag ctggtggctg ttccatcatg gtttttacct
     3241 cctgaattcg gatccctcga gcgatacagc gagtcgctct ttgtggaggt gacgattacc
     3301 taacaatcgg tcgattcgtt tgatgttatg ttttgttctc gctttggttg gcaggttacg
     3361 gccaagttcg gtaagagtga gagttttaca gtcaagtaat gcgtggcaag ccaacgttaa
     3421 gctgttgagt cgttttaagt gtaattcggg gcagaattgg taaagagagt cgtgtaaaat
     3481 atcgagttcg cacatcttgt tgtctgatta ttgatttttc gcgaaaccat ttaatcatat
     3541 gacaagatgt gtgtccacct taacttaatg atttttacca aaatcattag gggattcatc
     3601 agcgctgagt gtgtaaatta atttttatgc cgcagcgggc cagcaattct cgtgaatcat
     3661 cgcttaaacg gcctgatttc tgaatagcta tgcgtaagcg ggtgttgtct aacattctgc
     3721 gttcctcttt atcctgtctg aaccggctgc attaatgaat cggccaacgc gcggggagag
     3781 gcggtttgcg tattgggcgc tcttccgctt cctcg
//
