LOCUS       sequence-334624-        3645 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Donor DNA for TKIT knockin of SEP to GluA1 in mouse.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3645)
  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 3645)
  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-GluA1 TKIT donor
FEATURES             Location/Qualifiers
     source          1..3645
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(26..43)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(197..785)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(277..296)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(956..1747,1748..1816))
                     /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     1579..1598
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1817..1921)
                     /gene="bla"
                     /label=AmpR promoter
     CDS             2111..2821
                     /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(2138..2166)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     2761..2781
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
ORIGIN
        1 tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc
       61 tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg
      121 ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg
      181 ccgcattgct ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac
      241 gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg
      301 gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct
      361 ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg
      421 tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct
      481 gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac
      541 tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt
      601 tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc
      661 tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca
      721 ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat
      781 ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac
      841 gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt
      901 aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc
      961 aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg
     1021 cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg
     1081 ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc
     1141 cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta
     1201 ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg
     1261 ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct
     1321 ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta
     1381 gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg
     1441 ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga
     1501 ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt
     1561 gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca
     1621 ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt
     1681 cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt
     1741 ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga
     1801 aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat cagggttatt
     1861 gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc
     1921 gcacatttcc ccgactcgag aattctgacg tcattattat cagggccttc ctggtgttcc
     1981 attgctaagg ttggaccagg gcttcttttt cgccaatgta aaaaggaata tgccgtacat
     2041 ctttgccttt ttctgcaccg gttttctagg tgcggttgtg ggtgccaatt ctagtggtgg
     2101 cggtggctcg agtaaaggag aagaactttt cactggagtt gtcccaattc ttgttgaatt
     2161 agatggtgat gttaatgggc acaaattttc tgtcagtgga gagggtgaag gtgatgcaac
     2221 atacggaaaa cttaccctta aatttatttg cactactgga aaactacctg ttccttggcc
     2281 aacacttgtc actactttaa cttatggtgt tcaatgcttt tcaagatacc cagatcatat
     2341 gaaacggcat gactttttca agagtgccat gcccgaaggt tatgtacagg aaagaactat
     2401 atttttcaaa gatgacggga actacaagac acgtgctgaa gtcaagtttg aaggtgatac
     2461 ccttgttaat agaatcgagt taaaaggtat tgattttaaa gaagatggaa acattcttgg
     2521 acacaaattg gaatacaact ataacgatca ccaggtgtac atcatggcag acaaacaaaa
     2581 gaatggaatc aaagctaact tcaaaattag acacaacatt gaagatggag gcgttcaact
     2641 agcagaccat tatcaacaaa atactccaat tggcgatggg cccgtccttt taccagacaa
     2701 ccattacctg tttacaactt ctactctttc gaaagatccc aacgaaaaga gagaccacat
     2761 ggtccttctt gagtttgtaa cagctgctgg gattacacat ggcatggatg aactatacaa
     2821 aggtggcggt ggctcgggcg gaggtgggtc gggtggcggc ggatcaggcg gtggcggatc
     2881 cttccccaac aatatccaga taggtgagtg agaggggcag ctggggaggg actttgggga
     2941 tctggccagg attttctttt tcgttttttt ctttttcttt tttctttttt tttatttgga
     3001 ggggagattt ctcaacccat tctcagtcgg agctgttgtg cttagcaatg gaacaccacc
     3061 aaccatagat ttggtcttcc cacatcatgg tttttacctc ctgaattcgg atccctcgag
     3121 cgatacagcg agtcgctctt tgtggaggtg acgattacct aacaatcggt cgattcgttt
     3181 gatgttatgt tttgttctcg ctttggttgg caggttacgg ccaagttcgg taagagtgag
     3241 agttttacag tcaagtaatg cgtggcaagc caacgttaag ctgttgagtc gttttaagtg
     3301 taattcgggg cagaattggt aaagagagtc gtgtaaaata tcgagttcgc acatcttgtt
     3361 gtctgattat tgatttttcg cgaaaccatt taatcatatg acaagatgtg tgtccacctt
     3421 aacttaatga tttttaccaa aatcattagg ggattcatca gcgctgagtg tgtaaattaa
     3481 tttttatgcc gcagcgggcc agcaattctc gtgaatcatc gcttaaacgg cctgatttct
     3541 gaatagctat gcgtaagcgg gtgttgtcta acattctgcg ttcctcttta tcctgtctga
     3601 accggctgca ttaatgaatc ggccaacgcg cggggagagg cggtt
//
