LOCUS       sequence-211306-        1990 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Golden Gate Level 2 construct encoding Cas9-YFP, Zeocin resistance..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 1990)
  AUTHORS   Graff, Mock
  TITLE     Mock lab P. tricornutum plasmids
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 1990)
  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: 
            "Unpublished"
COMMENT     Sequence Label: pAGM4723:Ble:Cas9
FEATURES             Location/Qualifiers
     source          1..1990
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     CDS             979..999
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             join(1006..1008,1009..1011,1012..1725)
                     /codon_start=1
                     /product="enhanced YFP"
                     /label=EYFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1051..1072)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1312..1331)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1659..1680
                     /label=EGFP-C
                     /note="EGFP, forward primer"
ORIGIN
        1 aagaccgaga ttacactggc caatggagag attcggaagc gaccacttat cgaaacaaac
       61 ggagaaacag gagaaatcgt gtgggacaag ggtagggatt tcgcgacagt ccggaaggtc
      121 ctgtccatgc cgcaggtgaa catcgttaaa aagaccgaag tacagaccgg aggcttctcc
      181 aaggaaagta tcctcccgaa aaggaacagc gacaagctga tcgcacgcaa aaaagattgg
      241 gaccccaaga aatacggcgg attcgattct cctacagtcg cttacagtgt actggttgtg
      301 gccaaagtgg agaaagggaa gtctaaaaaa ctcaaaagcg tcaaggaact gctgggcatc
      361 acaatcatgg agcgatcaag cttcgaaaaa aaccccatcg actttctcga ggcgaaagga
      421 tataaagagg tcaaaaaaga cctcatcatt aagcttccca agtactctct ctttgagctt
      481 gaaaacggcc ggaaacgaat gctcgctagt gcgggcgagc tgcagaaagg taacgagctg
      541 gcactgccct ctaaatacgt taatttcttg tatctggcca gccactatga aaagctcaaa
      601 ggatctcccg aagataatga gcagaagcag ctgttcgtgg aacaacacaa acactacctt
      661 gatgagatca tcgagcaaat aagcgaattc tccaaaagag tgatcctcgc cgacgctaac
      721 ctcgataagg tgctttctgc ttacaataag cacagggata agcccatcag ggagcaggca
      781 gaaaacatta tccacttgtt tactctgacc aacttgggcg cgcctgcagc cttcaagtac
      841 ttcgacacca ccatagacag aaagcggtac acctctacaa aggaggtcct ggacgccaca
      901 ctgattcatc agtcaattac ggggctctat gaaacaagaa tcgacctctc tcagctcggt
      961 ggagacagca gggctgaccc caagaagaag aggaaggtgg gttcgatggt gagcaagggc
     1021 gaggagctgt tcaccggggt ggtgcccatc ctggtcgagc tggacggcga cgtaaacggc
     1081 cacaagttca gcgtgtccgg cgagggcgag ggcgatgcca cctacggcaa gctgaccctg
     1141 aagttcatct gcaccaccgg caagctgccc gtgccctggc ccaccctcgt gaccaccttc
     1201 ggctacggcc tgcagtgctt cgcccgctac cccgaccaca tgaagcagca cgacttcttc
     1261 aagtccgcca tgcccgaagg ctacgtccag gagcgcacca tcttcttcaa ggacgacggc
     1321 aactacaaga cccgcgccga ggtgaagttc gagggcgaca ccctggtgaa ccgcatcgag
     1381 ctgaagggca tcgacttcaa ggaggacggc aacatcctgg ggcacaagct ggagtacaac
     1441 tacaacagcc acaacgtcta tatcatggcc gacaagcaga agaacggcat caaggtgaac
     1501 ttcaagatcc gccacaacat cgaggacggc agcgtgcagc tcgccgacca ctaccagcag
     1561 aacaccccca tcggcgacgg ccccgtgctg ctgcccgaca accactacct gagctaccag
     1621 tccgccctga gcaaagaccc caacgagaag cgcgatcaca tggtcctgct ggagttcgtg
     1681 accgccgccg ggatcactct cggcatggac gagctgtaca agtgagcttc cttccttaaa
     1741 aatttaattt tcattagttg cagtcactcc gctttggttt cacagtcagg aataacacta
     1801 gctcgacttc accatggatg ccaatctcgc ctattcatgg tgtataaaag ttcaacatcc
     1861 aaagctagaa cttttggaaa gagaaagaat atccgaatag ggcacggcgt gccgtattgt
     1921 tggagtggac tagcagaaag tgaggaaggc acaggatgag ttttctcgag cgctactaga
     1981 ggatgcacat
//
