LOCUS       sequence-341867-        3961 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Donor DNA for TKIT knockin of SEP to NR1 in mouse.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3961)
  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 3961)
  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-GluN1 TKIT donor
FEATURES             Location/Qualifiers
     source          1..3961
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     139..158
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     392..409
                     /label=L4440
                     /note="L4440 vector, forward primer"
     CDS             complement(1480..2190)
                     /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(1520..1540)
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     primer_bind     2135..2163
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     promoter        2475..2579
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2580..2648,2649..3440)
                     /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     complement(2798..2817)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      join(3611..3961,1..238)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct
       61 tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg
      121 cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca
      181 cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa
      241 cgccagcaat gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt
      301 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga
      361 taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga
      421 gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagccggtt
      481 cagacaggat aaagaggaac gcagaatgtt agacaacacc cgcttacgca tagctattca
      541 gaaatcaggc cgtttaagcg atgattcacg agaattgctg gcccgctgcg gcataaaaat
      601 taatttacac actcagcgct gatgaatccc ctaatgattt tggtaaaaat cattaagtta
      661 aggtggacac acatcttgtc atatgattaa atggtttcgc gaaaaatcaa taatcagaca
      721 acaagatgtg cgaactcgat attttacacg actctcttta ccaattctgc cccgaattac
      781 acttaaaacg actcaacagc ttaacgttgg cttgccacgc attacttgac tgtaaaactc
      841 tcactcttac cgaacttggc cgtaacctgc caaccaaagc gagaacaaaa cataacatca
      901 aacgaatcga ccgattgtta ggtaatcgtc acctccacaa agagcgactc gctgtatcgc
      961 tcgagggatc cgaattccgc gcgggtgttc gagcagcgct caccaatgat agtcacatga
     1021 cagcacacat ggtcctctga ggaccagtcc agggggaacc agcagacaca gacatttgtc
     1081 ccaggccttt ccagggacca gttggcttgg ggtggtctgg taaggagact gagaaggacc
     1141 agggaaggat cgaatgaaac tggggcagtg gaggatggtg gaggagcagg ggagagagga
     1201 agtagtggtc acctggctag agatgaggtc ctcacacact gacagggcca tctgtatggc
     1261 gttgggcttg tgggtgacag aagtggcgtt gagctgtatc ttccaagagc cgtgtcgctt
     1321 attggcctgg tttactgcct cgcggaacat ctgctcgtgc ttgcgcgtgc tcagcaccgc
     1381 gccgatgttg acaatcttgg ggtcgcaggc agcgcgggat ccgccaccgc ctgatccgcc
     1441 gccacccgac ccacctccgc ccgagccacc gccaccaagc ttgtatagtt catccatgcc
     1501 atgtgtaatc ccagcagctg ttacaaactc aagaaggacc atgtggtctc tcttttcgtt
     1561 gggatctttc gaaagagtag aagttgtaaa caggtaatgg ttgtctggta aaaggacggg
     1621 cccatcgcca attggagtat tttgttgata atggtctgct agttgaacgc ctccatcttc
     1681 aatgttgtgt ctaattttga agttagcttt gattccattc ttttgtttgt ctgccatgat
     1741 gtacacctgg tgatcgttat agttgtattc caatttgtgt ccaagaatgt ttccatcttc
     1801 tttaaaatca atacctttta actcgattct attaacaagg gtatcacctt caaacttgac
     1861 ttcagcacgt gtcttgtagt tcccgtcatc tttgaaaaat atagttcttt cctgtacata
     1921 accttcgggc atggcactct tgaaaaagtc atgccgtttc atatgatctg ggtatcttga
     1981 aaagcattga acaccataag ttaaagtagt gacaagtgtt ggccaaggaa caggtagttt
     2041 tccagtagtg caaataaatt taagggtaag ttttccgtat gttgcatcac cttcaccctc
     2101 tccactgaca gaaaatttgt gcccattaac atcaccatct aattcaacaa gaattgggac
     2161 aactccagtg aaaagttctt ctcctttact tctagagcca ccgccaccgg cagcgcgggc
     2221 gaaggagcag gagaaaagca gggcgaatgt cagcaggtgc atggtgctca tgagcctcgg
     2281 gcacagcggg cctggttctg ggttgcgcga gcgcgaccac ctcgcgatgt acggaccccc
     2341 gggcccccgc gcggtcaagc acccgcggat tcttcaggag cccagggcga gcgcggcggg
     2401 tgctgtccga ggtgctgaag cgtattggtc tgttccatct agtcagtgtg gaattctcga
     2461 gtcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt
     2521 atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagta
     2581 tgagtattca acatttccgt gtcgccctta ttcccttttt tgcggcattt tgccttcctg
     2641 tttttgctca cccagaaacg ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac
     2701 gagtgggtta catcgaactg gatctcaaca gcggtaagat ccttgagagt tttcgccccg
     2761 aagaacgttt tccaatgatg agcactttta aagttctgct atgtggcgcg gtattatccc
     2821 gtattgacgc cgggcaagag caactcggtc gccgcataca ctattctcag aatgacttgg
     2881 ttgagtactc accagtcaca gaaaagcatc ttacggatgg catgacagta agagaattat
     2941 gcagtgctgc cataaccatg agtgataaca ctgcggccaa cttacttctg acaacgatcg
     3001 gaggaccgaa ggagctaacc gcttttttgc acaacatggg ggatcatgta actcgccttg
     3061 atcgttggga accggagctg aatgaagcca taccaaacga cgagcgtgac accacgatgc
     3121 ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg cgaactactt actctagctt
     3181 cccggcaaca attaatagac tggatggagg cggataaagt tgcaggacca cttctgcgct
     3241 cggcccttcc ggctggctgg tttattgctg ataaatctgg agccggtgag cgtgggtctc
     3301 gcggtatcat tgcagcactg gggccagatg gtaagccctc ccgtatcgta gttatctaca
     3361 cgacggggag tcaggcaact atggatgaac gaaatagaca gatcgctgag ataggtgcct
     3421 cactgattaa gcattggtaa ctgtcagacc aagtttactc atatatactt tagattgatt
     3481 taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga
     3541 ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca
     3601 aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac
     3661 caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg
     3721 taactggctt cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag
     3781 gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac
     3841 cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt
     3901 taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg
     3961 a
//
