LOCUS       P527_POU4F2-p2A-mNeonGreen-h2b        5691 bp    DNA     circular SYN 22-APR-2025
DEFINITION  Azenta NGS; inserts a p2A-mNeonGreen-h2b fusion at the endogenous
            POU4F2 gene.
ACCESSION
VERSION     .
KEYWORDS    .
SOURCE      .
  ORGANISM  .
COMMENT     Annotated with pLannotate v1.2.0.
FEATURES             Location/Qualifiers
     misc_feature    202..267
                     /note="Geneious type: P2A"
                     /note="Derived using Geneious Prime 2025.0.3 'Annotate
                     from Database' based on nucleotide similarity"
                     /standard_name="p2A"
     source          268..978
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
                     /note="Derived using Geneious Prime 2025.0.3 'Annotate
                     from Database' based on nucleotide similarity"
                     /note="Geneious type: GFP"
     misc_feature    976..1352
                     /note="Geneious type: NLS"
                     /note="Derived using Geneious Prime 2025.0.3 'Annotate
                     from Database' based on nucleotide similarity"
                     /standard_name="H2B NLS"
     misc_feature    1354..2284
                     /note="Geneious type: homology arm"
                     /note="Derived using Geneious Prime 2025.1.1 'Annotate
                     from Database' based on nucleotide similarity"
                     /standard_name="POU4F2 RHA"
     regulatory      2420..2670
                     /note="Geneious type: promoter"
                     /note="Derived using Geneious Prime 2025.0.3 'Annotate
                     from Database' based on nucleotide similarity"
                     /standard_name="U6 promoter"
     misc_feature    2671..2690
                     /note="Geneious type: gRNA"
                     /note="Derived using Geneious Prime 2025.0.3 'Annotate
                     from Database' based on nucleotide similarity"
                     /standard_name="BRN3B gRNA T2"
     misc_feature    2691..2772
                     /note="Geneious type: grna scaffold"
                     /note="Derived using Geneious Prime 2025.0.3 'Annotate
                     from Database' based on nucleotide similarity"
                     /standard_name="gRNA scaffold V1"
     rep_origin      complement(3038..3626)
     misc_RNA        complement(3038..3590)
                     /note="RNAII transcript (complementary strand)"
                     /note="Derived using Geneious Prime 2025.0.3 'Annotate
                     from Database' based on nucleotide similarity"
     misc_RNA        3480..3587
                     /note="RNAI transcript"
                     /note="Derived using Geneious Prime 2025.0.3 'Annotate
                     from Database' based on nucleotide similarity"
     misc_feature    complement(3797..4657)
                     /note="Geneious type: Marker"
                     /note="Derived using Geneious Prime 2025.0.3 'Annotate
                     from Database' based on nucleotide similarity"
                     /standard_name="amp marker"
     regulatory      complement(4658..4762)
                     /note="Geneious type: promoter"
                     /note="Promoter for ampicillin resistance gene
                     (b-lactamase)"
                     /regulatory_class="promoter"
                     /note="Derived using Geneious Prime 2024.0.5 'Annotate
                     from Database' based on nucleotide similarity"
                     /note="Derived using Geneious Prime 2025.0.3 'Annotate
                     from Database' based on nucleotide similarity"
                     /standard_name="amp"
     misc_feature    5082..200
                     /note="Geneious type: homology arm"
                     /note="Derived using Geneious Prime 2025.0.3 'Annotate
                     from Database' based on nucleotide similarity"
                     /standard_name="POU4F2 LHA"
     misc_feature    5098..1356
                     /note="Geneious type: ORF"
                     /standard_name="ORF 10 (frame 1)"
ORIGIN      
        1 gcggagaaga agcgcaagcg cacgtccatc gctgcgccag agaagcgctc gctcgaagcc
       61 tactttgcca ttcagcctcg gccctcctct gaaaagatcg ccgccatcgc ggagaagctg
      121 gacctgaaga aaaacgtggt gcgcgtctgg ttctgcaacc agaggcagaa acagaaaaga
      181 atgaaatatt ccgccggcat tgggagcgga gctactaact tcagcctgct gaagcaggct
      241 ggagacgtgg aggagaaccc tggacctatg gtgagcaagg gcgaggagga taacatggcc
      301 tctctcccag cgacacatga gttacacatc tttggctcca tcaacggtgt ggactttgac
      361 atggtgggtc agggcaccgg caatccaaat gatggttatg aggagttaaa cctgaagtcc
      421 accaagggtg acctccagtt ctccccctgg attctggtcc ctcatatcgg gtatggcttc
      481 catcagtacc tgccctaccc tgacgggatg tcgcctttcc aggccgccat ggtagatggc
      541 tccggatacc aagtccatcg cacaatgcag tttgaagatg gtgcctccct tactgttaac
      601 taccgctaca cctacgaggg aagccacatc aaaggagagg cccaggtgaa ggggactggt
      661 ttccctgctg acggtcctgt gatgaccaac tcgctgaccg ctgcggactg gtgcaggtcg
      721 aagaagactt accccaacga caaaaccatc atcagtacct ttaagtggag ttacaccact
      781 ggaaatggca agcgctaccg gagcactgcg cggaccacct acacctttgc caagccaatg
      841 gcggctaact atctgaagaa ccagccgatg tacgtgttcc gtaagacgga gctcaagcac
      901 tccaagaccg agctcaactt caaggagtgg caaaaggcct ttaccgatgt gatgggcatg
      961 gacgagctgt acaagatgcc tgaaccctct agttctgctc cagcccctaa aaagggttct
     1021 aagaaggcta tcactaaggc gcagaagaag gatggtaaga agcgtaagcg cagccgcaag
     1081 gagagctatt ctatctatgt gtacaaggtt ctgaagcagg tccaccccga caccggcatc
     1141 tcatccaagg ccatggggat catgaactcc ttcgtcaacg acatcttcga gcgcatcgcg
     1201 ggcgaggctt ctcgcctggc tcactacaat aagcgctcga ccatcacctc cagggagatt
     1261 cagacggctg tgcgcctgct gctgcctggg gagctggcta agcatgctgt gtccgagggc
     1321 actaaggcag ttaccaagta cactagctct aaatagaaga ctcttggcct ctccagagac
     1381 gcccctttcc tcgtccgctc ttttctctcc tctcttctgc ctcttttcac ttttggcgac
     1441 tagaaacaat tccagtaaat gtgaatctcg acaaatcgag gactgaagag ggagcgaacg
     1501 agcgaacaac tgagcccaag ccggtgagaa tgtgaaacag tttctcaaag gaaagaataa
     1561 caaaagatgg tatttgtctg ttgtagcaaa gttgtccctt tgaaccccac ctcggcttct
     1621 tcagaggaag tgtggagatg gctgtttgca ggaaggcaga cgagacagtg tttaaaaagt
     1681 ccacaagaat gatcaagtaa gatttgtttt tattcttaca gacatcaccc gtgttcaagt
     1741 ttaaaagtac actttgcaac tatttttcag aaatagaaat tgattcagga ctaaaacttt
     1801 aaactagagt tgatgcttaa tgtgatagag acatctctaa agtattttga attttaaaaa
     1861 aagatggcag attttctgca tttacactgt atattatata tatattttta ttgtggttct
     1921 tacccccttt tccttctctg aagtgttaat gcttaagaaa agagttgcgc ctgctgtgtt
     1981 cactgatctt gaaagctatt attagattat tgcagaacaa ccctctgtaa attattaatt
     2041 tatctctcta gcaacttaat tttgtgcaca ttctaattaa ttaaacttct tccgtctaaa
     2101 aaaagtgggg gaaatgtata gctagtaacg ttcaaaaaat tttgtttgat gagtttaccg
     2161 aatttttaca gctttcctcc tatactgtgt tccttttgac ccatttgtat attctcactt
     2221 gaatgaagat tgtttttttc tttgttttta ctggtagtgt tctgatttgt gagtcgacac
     2281 tcagtttggt accaagcttt gcgcggtgtg gaaagtcccc aggctcccca ggcaggcaga
     2341 agtatgcaaa gcatgcatct caattagtca gcaaccagcc taggcgccta gggaattcgc
     2401 ggccgccttt tgctcacatg tgagggccta tttcccatga ttccttcata tttgcatata
     2461 cgatacaagg ctgttagaga gataattgga attaatttga ctgtaaacac aaagatatta
     2521 gtacaaaata cgtgacgtag aaagtaataa tttcttgggt agtttgcagt tttaaaatta
     2581 tgttttaaaa tggactatca tatgcttacc gtaacttgaa agtatttcga tttcttggct
     2641 ttatatatct tgtggaaagg acgaaacacc gccggcattt agaagactct gttttagagc
     2701 tagaaatagc aagttaaaat aaggctagtc cgttatcaac ttgaaaaagt ggcaccgagt
     2761 cggtgctttt ttggcggccg cgaccccaac tggggtaacc tttgagttct ctcagttggg
     2821 gtccataggg ataacagggt aatttgatat cgcgctcttc cgcttcctcg ctcactgact
     2881 cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac
     2941 ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa
     3001 aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg
     3061 acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa
     3121 gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc
     3181 ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac
     3241 gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac
     3301 cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg
     3361 taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt
     3421 atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa
     3481 cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct
     3541 cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga
     3601 ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg
     3661 ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct
     3721 tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt
     3781 aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc
     3841 tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg
     3901 gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag
     3961 atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt
     4021 tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag
     4081 ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt
     4141 ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca
     4201 tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg
     4261 ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat
     4321 ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta
     4381 tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca
     4441 gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct
     4501 taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat
     4561 cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa
     4621 agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt
     4681 gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa
     4741 ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac gtcgacggat
     4801 cgggagatct cccgatcccc tatggtgcac tctcagtaca atctgctctg atgccgcata
     4861 gttaagccag tatctgctcc ctgcttgtgt gttggaggtc gctgagtagt gcgcgagcaa
     4921 aatttaagct acaacaaggc aaggcttgac cgacaattgc atgaagaatc tgcttagggt
     4981 taggcgtttt gtgctgcttc gtagggataa cagggtaatt catgggtgcc agggcgtgcc
     5041 cttgggctcc ccgggcgcgt tcgatgtacg ggccagatat acgccaccta ccacactatg
     5101 aataccatcc cgtgcacgtc ggccgcctct tcttcatcgg tgcccatctc gcacccttcc
     5161 gcgttggcgg gcacgcacca ccaccaccac catcaccacc accaccacca ccaaccgcac
     5221 caggcgctgg agggcgagct gctggagcac ctgagtcccg ggctggccct gggcgctatg
     5281 gcgggccccg acggcgctgt ggtgtccacg ccggctcacg cgccgcacat ggccaccatg
     5341 aaccccatgc accaagcagc gctcagcatg gcccacgcgc acgggctgcc gtcgcacatg
     5401 ggctgcatga gcgacgtgga cgccgacccg cgggacctgg aggcattcgc cgagcgcttc
     5461 aagcagcgac gcatcaagct gggggtgacc caggcagatg tgggctccgc gctggccaac
     5521 ctcaagatcc ccggcgtggg ctcgcttagc cagagcacca tctgcaggtt cgagtccctc
     5581 acactgtccc acaataatat gatcgcgctc aaacccatcc tgcaggcatg gctcgaggag
     5641 gccgagaagt cccaccgcga gaagctcacc aagcctgaac tcttcaatgg c
//
