LOCUS       sequence-140827-        3272 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Middle entry clone with photoconvertible eos conjugated to a nuclear
            localization signal.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3272)
  AUTHORS   Curran K, Lister JA, Kunkel GR, Prendergast A, Parichy DM, Raible DW
  TITLE     Interplay between Foxd3 and Mitf regulates cell fate plasticity in 
            the zebrafish neural crest.
  JOURNAL   Dev Biol. 2010 Aug 1;344(1):107-18. doi: 
            10.1016/j.ydbio.2010.04.023. Epub 2010 May 9.
  PUBMED    20460180
REFERENCE   2  (bases 1 to 3272)
  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"; doi: 
            "10.1016/j.ydbio.2010.04.023"; journalName: "Dev Biol"; date: 
            "2010-08-1- 1"; volume: "344"; issue: "1"; pages: "107-18"
COMMENT     Sequence Label: pME nls-eos
FEATURES             Location/Qualifiers
     source          1..3272
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     92..109
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(268..295)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(387..473)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     522..544
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     536..553
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     537..553
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    569..668
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     regulatory      671..680
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             683..703
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             710..1390
                     /codon_start=1
                     /product="green-to-red photoswitchable fluorescent protein 
                     (Wiedenmann et al., 2004)"
                     /label=EosFP
                     /translation="MSAIKPDMKINLRMEGNVNGHHFVIDGDGTGKPFEGKQSMDLEVK
                     EGGPLPFAFDILTTAFHYGNRVFAEYPDHIQDYFKQSFPKGYSWERSLTFEDGGICIAR
                     NDITMEGDTFYNKVRFHGVNFPANGPVMQKKTLKWEPSTEKMYVRDGVLTGDITMALLL
                     EGNAHYRCDFRTTYKAKEKGVKLPGYHFVDHCIEILSHDKDYNKVKLYEHAVAHSGLPD
                     NARR"
     protein_bind    complement(1397..1496)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1513..1532)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1514..1532)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1537..1553)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1537..1553)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1655..1674)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             1666..2475
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP
                     DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA
                     FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
                     FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
                     RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     complement(1733..1752)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      2622..3210
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3111..3130
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga
       61 taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga
      121 gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctggca
      181 cgacaggttt cccgactgga aagcgggcag tgagcgcaac gcaattaata cgcgtaccgc
      241 tagccaggaa gagtttgtag aaacgcaaaa aggccatccg tcaggatggc cttctgctta
      301 gtttgatgcc tggcagttta tggcgggcgt cctgcccgcc accctccggg ccgttgcttc
      361 acaacgttca aatccgctcc cggcggattt gtcctactca ggagagcgtt caccgacaaa
      421 caacagataa aacgaaaggc ccagtcttcc gactgagcct ttcgttttat ttgatgcctg
      481 gcagttccct actctcgcgt taacgctagc atggatgttt tcccagtcac gacgttgtaa
      541 aacgacggcc agtcttaagc tcgggcccca aataatgatt ttattttgac tgatagtgac
      601 ctgttcgttg caacaaattg atgagcaatg cttttttata atgccaactt tgtacaaaaa
      661 agcaggctgc gccaccatgg ctccaaagaa gaagcgtaag gtagaattca tgagtgcgat
      721 taagccagac atgaagatca acctccgtat ggaaggcaac gtaaacgggc accactttgt
      781 gatcgacgga gatggtacag gcaagccttt tgagggaaaa cagagtatgg atcttgaagt
      841 caaagagggc ggacctctgc cttttgcctt tgatatcctg accactgcat tccattacgg
      901 caacagggta ttcgccgaat atccagacca catacaagac tattttaagc agtcgtttcc
      961 taaggggtat tcgtgggaac gaagcttgac tttcgaagac gggggcattt gcattgccag
     1021 aaacgacata acaatggaag gggacacttt ctataataaa gttcgatttc acggtgtaaa
     1081 ctttcccgcc aatggtccag ttatgcagaa gaagacgctg aaatgggagc cctccactga
     1141 gaaaatgtat gtgcgtgatg gagtgctgac gggtgatatt accatggctt tgttgcttga
     1201 aggaaatgcc cattaccgat gtgacttcag aactacttac aaagctaagg agaagggtgt
     1261 caagttacca ggctaccact ttgtggacca ctgcattgag attttaagcc atgacaaaga
     1321 ttacaacaag gttaagctgt atgagcatgc tgttgctcat tctggattgc ctgacaatgc
     1381 cagacgatag ggatccaccc agctttcttg tacaaagttg gcattataag aaagcattgc
     1441 ttatcaattt gttgcaacga acaggtcact atcagtcaaa ataaaatcat tatttgccat
     1501 ccagctgata tcccctatag tgagtcgtat tacatggtca tagctgtttc ctggcagctc
     1561 tggcccgtgt ctcaaaatct ctgatgttac attgcacaag ataaaaatat atcatcatga
     1621 acaataaaac tgtctgctta cataaacagt aatacaaggg gtgttatgag ccatattcaa
     1681 cgggaaacgt cgaggccgcg attaaattcc aacatggatg ctgatttata tgggtataaa
     1741 tgggctcgcg ataatgtcgg gcaatcaggt gcgacaatct atcgcttgta tgggaagccc
     1801 gatgcgccag agttgtttct gaaacatggc aaaggtagcg ttgccaatga tgttacagat
     1861 gagatggtca gactaaactg gctgacggaa tttatgcctc ttccgaccat caagcatttt
     1921 atccgtactc ctgatgatgc atggttactc accactgcga tccccggaaa aacagcattc
     1981 caggtattag aagaatatcc tgattcaggt gaaaatattg ttgatgcgct ggcagtgttc
     2041 ctgcgccggt tgcattcgat tcctgtttgt aattgtcctt ttaacagcga tcgcgtattt
     2101 cgtctcgctc aggcgcaatc acgaatgaat aacggtttgg ttgatgcgag tgattttgat
     2161 gacgagcgta atggctggcc tgttgaacaa gtctggaaag aaatgcataa acttttgcca
     2221 ttctcaccgg attcagtcgt cactcatggt gatttctcac ttgataacct tatttttgac
     2281 gaggggaaat taataggttg tattgatgtt ggacgagtcg gaatcgcaga ccgataccag
     2341 gatcttgcca tcctatggaa ctgcctcggt gagttttctc cttcattaca gaaacggctt
     2401 tttcaaaaat atggtattga taatcctgat atgaataaat tgcagtttca tttgatgctc
     2461 gatgagtttt tctaatcaga attggttaat tggttgtaac actggcagag cattacgctg
     2521 acttgacggg acggcgcaag ctcatgacca aaatccctta acgtgagtta cgcgtcgttc
     2581 cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg
     2641 cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg
     2701 gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca
     2761 aatactgttc ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg
     2821 cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg
     2881 tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga
     2941 acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac
     3001 ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat
     3061 ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc
     3121 tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga
     3181 tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc
     3241 ctggcctttt gctggccttt tgctcacatg tt
//
