LOCUS       sequence-334640-        3109 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Donor DNA for TKIT knockin of SEP to GluA1 in rat.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3109)
  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 3109)
  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: pHFS:SEP-rGluA1 TKIT donor
FEATURES             Location/Qualifiers
     source          1..3109
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     433..452
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     686..703
                     /label=L4440
                     /note="L4440 vector, forward primer"
     CDS             complement(1039..1749)
                     /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(1079..1099)
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     primer_bind     1694..1722
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     promoter        1917..2021
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2022..2090,2091..2882)
                     /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(2240..2259)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      join(3053..3109,1..532)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg
       61 cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt taggccacca
      121 cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc
      181 tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga
      241 taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac
      301 gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga
      361 agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag
      421 ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg
      481 acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag
      541 caatgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc
      601 tgcgttatcc cctgattctg tggataaccg tattaccgcc tttgagtgag ctgataccgc
      661 tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg aagagcgccc
      721 agggaagacc aaatctatgg ttggggcatt tgacgagcat gaaaccccct ttcccagact
      781 ggcgacccaa gtcttcctcc catcctggtt ttagattgct aagcacaaca gctccgactg
      841 agaatgggtt gagaaatctc ccctcaaaaa aagggggaaa aaaaaagaaa gaaagaaaat
      901 cctggccaga tccccaaagt ccctccccag ctgcccctct cactcaccta tctggatatt
      961 gttggggaaa ttggcggatc cgccaccgcc tgatccgccg ccacccgacc cacctccgcc
     1021 cgagccaccg ccaccaagct tgtatagttc atccatgcca tgtgtaatcc cagcagctgt
     1081 tacaaactca agaaggacca tgtggtctct cttttcgttg ggatctttcg aaagagtaga
     1141 agttgtaaac aggtaatggt tgtctggtaa aaggacgggc ccatcgccaa ttggagtatt
     1201 ttgttgataa tggtctgcta gttgaacgcc tccatcttca atgttgtgtc taattttgaa
     1261 gttagctttg attccattct tttgtttgtc tgccatgatg tacacctggt gatcgttata
     1321 gttgtattcc aatttgtgtc caagaatgtt tccatcttct ttaaaatcaa taccttttaa
     1381 ctcgattcta ttaacaaggg tatcaccttc aaacttgact tcagcacgtg tcttgtagtt
     1441 cccgtcatct ttgaaaaata tagttctttc ctgtacataa ccttcgggca tggcactctt
     1501 gaaaaagtca tgccgtttca tatgatctgg gtatcttgaa aagcattgaa caccataagt
     1561 taaagtagtg acaagtgttg gccaaggaac aggtagtttt ccagtagtgc aaataaattt
     1621 aagggtaagt tttccgtatg ttgcatcacc ttcaccctct ccactgacag aaaatttgtg
     1681 cccattaaca tcaccatcta attcaacaag aattgggaca actccagtga aaagttcttc
     1741 tcctttactt ctagagccac cgccaccact agaattggca cccacaaccg cacctagaaa
     1801 accggtgcag aaaaaggcaa agatgtacgg catattcctt tttacattgg cgaaaaagaa
     1861 gccctggtcc aacccatgct cgtcaaatgc cacgaggctc gagtcgggga aatgtgcgcg
     1921 gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat
     1981 aaccctgata aatgcttcaa taatattgaa aaaggaagag tatgagtatt caacatttcc
     2041 gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct cacccagaaa
     2101 cgctggtgaa agtaaaagat gctgaagatc agttgggtgc acgagtgggt tacatcgaac
     2161 tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgt tttccaatga
     2221 tgagcacttt taaagttctg ctatgtggcg cggtattatc ccgtattgac gccgggcaag
     2281 agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtac tcaccagtca
     2341 cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct gccataacca
     2401 tgagtgataa cactgcggcc aacttacttc tgacaacgat cggaggaccg aaggagctaa
     2461 ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg gaaccggagc
     2521 tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgtagca atggcaacaa
     2581 cgttgcgcaa actattaact ggcgaactac ttactctagc ttcccggcaa caattaatag
     2641 actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt ccggctggct
     2701 ggtttattgc tgataaatct ggagccggtg agcgtgggtc tcgcggtatc attgcagcac
     2761 tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg agtcaggcaa
     2821 ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt aagcattggt
     2881 aactgtcaga ccaagtttac tcatatatac tttagattga tttaaaactt catttttaat
     2941 ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg
     3001 agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc
     3061 ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgct
//
