LOCUS       sequence-334641-        4060 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Donor DNA for TKIT knockin of SEP to NR2B in mouse.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4060)
  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 4060)
  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-GluN2B TKIT donor
FEATURES             Location/Qualifiers
     source          1..4060
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(13..30)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(184..772)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(264..283)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(943..1734,1735..1803))
                     /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     1566..1585
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1804..1908)
                     /gene="bla"
                     /label=AmpR promoter
     CDS             2169..2879
                     /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(2196..2224)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     2819..2839
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
ORIGIN
        1 ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt
       61 atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa
      121 gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cattgctggc
      181 gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag
      241 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt
      301 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg
      361 aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg
      421 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg
      481 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac
      541 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg
      601 gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt
      661 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg
      721 tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc
      781 tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt
      841 ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt
      901 taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag
      961 tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt
     1021 cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg caatgatacc
     1081 gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc
     1141 cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg
     1201 ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac
     1261 aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg
     1321 atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc
     1381 tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact
     1441 gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc
     1501 aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat
     1561 acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc
     1621 ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac
     1681 tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa
     1741 aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact
     1801 catactcttc ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg
     1861 atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg
     1921 actcgagaat tcccctgtga aagggtcatt caatggattt tctagcattc agacacttca
     1981 aggatgcgtt ccacaacgcg ggtcacgttt taaataaatt tcttccttct ctttttcaga
     2041 gagtcgacga gctgaagatg aagcccagcg cagagtgctg ttctcccaag ttctggttgg
     2101 tgttggccgt cttggccgta tcgggcagca aagctcgttc ccaaaagagc ggtggcggtg
     2161 gctctagaag taaaggagaa gaacttttca ctggagttgt cccaattctt gttgaattag
     2221 atggtgatgt taatgggcac aaattttctg tcagtggaga gggtgaaggt gatgcaacat
     2281 acggaaaact tacccttaaa tttatttgca ctactggaaa actacctgtt ccttggccaa
     2341 cacttgtcac tactttaact tatggtgttc aatgcttttc aagataccca gatcatatga
     2401 aacggcatga ctttttcaag agtgccatgc ccgaaggtta tgtacaggaa agaactatat
     2461 ttttcaaaga tgacgggaac tacaagacac gtgctgaagt caagtttgaa ggtgataccc
     2521 ttgttaatag aatcgagtta aaaggtattg attttaaaga agatggaaac attcttggac
     2581 acaaattgga atacaactat aacgatcacc aggtgtacat catggcagac aaacaaaaga
     2641 atggaatcaa agctaacttc aaaattagac acaacattga agatggaggc gttcaactag
     2701 cagaccatta tcaacaaaat actccaattg gcgatgggcc cgtcctttta ccagacaacc
     2761 attacctgtt tacaacttct actctttcga aagatcccaa cgaaaagaga gaccacatgg
     2821 tccttcttga gtttgtaaca gctgctggga ttacacatgg catggatgaa ctatacaagc
     2881 ttggtggcgg tggctcgggc ggaggtgggt cgggtggcgg cggatcaggc ggtggcggat
     2941 cccaaaagag cgcccccagc atcggcatcg ctgtcatcct cgtgggcact tccgacgaag
     3001 tggccataaa agatgcccac gagaaagatg acttccatca tctctcagta gttccccggg
     3061 tggagctggt agccatgaac gagactgacc caaagagcat aatcacccgc atctgcgatc
     3121 ttatgtctga ccggaagatc cagggggtgg tgttcgcgga tgacacggac caggaagcca
     3181 tcgcccagat cctcgatttc atttctgctc agactctcac ccccatcctg ggcatccatg
     3241 ggggctcatc tatgataatg gcagataagg taaaaaggag gctattccaa aaagcaaatg
     3301 atgcattggt ttttaaggca aaactaagta gttaaagata gacgtgcacc agagcacatc
     3361 gatgagtgag cgaatggatg ggaactagtc agatagttag gagatgaaag tgaaaatgtg
     3421 atagtggtgt agaaaggtct acagcaatgc cgctcaacct gtgggtcgtg acccattcag
     3481 gaagaacctc gtctccgtct gcgggaattc ggatccctcg agcgatacag cgagtcgctc
     3541 tttgtggagg tgacgattac ctaacaatcg gtcgattcgt ttgatgttat gttttgttct
     3601 cgctttggtt ggcaggttac ggccaagttc ggtaagagtg agagttttac agtcaagtaa
     3661 tgcgtggcaa gccaacgtta agctgttgag tcgttttaag tgtaattcgg ggcagaattg
     3721 gtaaagagag tcgtgtaaaa tatcgagttc gcacatcttg ttgtctgatt attgattttt
     3781 cgcgaaacca tttaatcata tgacaagatg tgtgtccacc ttaacttaat gatttttacc
     3841 aaaatcatta ggggattcat cagcgctgag tgtgtaaatt aatttttatg ccgcagcggg
     3901 ccagcaattc tcgtgaatca tcgcttaaac ggcctgattt ctgaatagct atgcgtaagc
     3961 gggtgttgtc taacattctg cgttcctctt tatcctgtct gaaccggctg cattaatgaa
     4021 tcggccaacg cgcggggaga ggcggtttgc gtattgggcg
//
