LOCUS       sequence-263459-        3239 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  full-length of photoconvertible kaede with stop codon in pmE Gateway
            vector.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3239)
  AUTHORS   Shin M, Nozaki T, Idrizi F, Isogai S, Ogasawara K, Ishida K, Yuge S,
            Roscoe B, Wolfe SA, Fukuhara S, Mochizuki N, Deguchi T, Lawson ND
  TITLE     Valves Are a Conserved Feature of the Zebrafish Lymphatic System.
  JOURNAL   Dev Cell. 2019 Nov 4;51(3):374-386.e5. doi: 
            10.1016/j.devcel.2019.08.019. Epub 2019 Sep 26.
  PUBMED    31564611
REFERENCE   2  (bases 1 to 3239)
  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.devcel.2019.08.019"; journalName: "Dev Cell"; date: 
            "2019-11-4- 4"; volume: "51"; issue: "3"; pages: "374-386.e5"
COMMENT     Sequence Label: pME-Kaede
FEATURES             Location/Qualifiers
     source          1..3239
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     33..52
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     286..303
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(462..489)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(581..667)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     716..738
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     730..747
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     731..747
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    763..862
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             880..1557
                     /codon_start=1
                     /product="Kaede green-to-red photoconvertible fluorescent 
                     protein (Ando et al., 2002)"
                     /label=Kaede
                     /translation="MSLIKPEMKIKLLMEGNVNGHQFVIEGDGKGHPFEGKQSMDLVVK
                     EGAPLPFAYDILTTAFHYGNRVFAKYPDHIPDYFKQSFPKGFSWERSLMFEDGGVCIAT
                     NDITLKGDTFFNKVRFDGVNFPPNGPVMQKKTLKWEASTEKMYLRDGVLTGDITMALLL
                     KGDVHYRCDFRTTYKSRQEGVKLPGYHFVDHCISILRHDKDYNEVKLYEHAVAHSGLPD
                     NVK"
     protein_bind    complement(1558..1657)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1674..1693)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1675..1693)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1698..1714)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1698..1714)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1816..1835)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             1827..2636
                     /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(1894..1913)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      join(2783..3239,1..132)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct
       61 gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg
      121 agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct
      181 tttgctcaca tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc
      241 tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc
      301 gaggaagcgg aagagcgccc aatacgcaaa ccgcctctcc ccgcgcgttg gccgattcat
      361 taatgcagct ggcacgacag gtttcccgac tggaaagcgg gcagtgagcg caacgcaatt
      421 aatacgcgta ccgctagcca ggaagagttt gtagaaacgc aaaaaggcca tccgtcagga
      481 tggccttctg cttagtttga tgcctggcag tttatggcgg gcgtcctgcc cgccaccctc
      541 cgggccgttg cttcacaacg ttcaaatccg ctcccggcgg atttgtccta ctcaggagag
      601 cgttcaccga caaacaacag ataaaacgaa aggcccagtc ttccgactga gcctttcgtt
      661 ttatttgatg cctggcagtt ccctactctc gcgttaacgc tagcatggat gttttcccag
      721 tcacgacgtt gtaaaacgac ggccagtctt aagctcgggc cccaaataat gattttattt
      781 tgactgatag tgacctgttc gttgcaacaa attgatgagc aatgcttttt tataatgcca
      841 actttgtaca aaaaagcagg ctcgctaccg gtcgccacca tgagtctgat taaaccagaa
      901 atgaagatca agctgcttat ggaaggcaat gtaaacgggc accagtttgt tattgaggga
      961 gatggaaaag gccatccttt tgagggaaaa cagagtatgg accttgtagt caaagaaggc
     1021 gcacctctcc cttttgccta cgatatcttg acaacagcat tccattatgg taacagggtt
     1081 tttgctaaat acccagacca tataccagac tacttcaagc agtcgtttcc caaagggttt
     1141 tcttgggagc gaagcctgat gttcgaggac gggggcgttt gcatcgctac aaatgacata
     1201 acactgaaag gagacacttt ttttaacaaa gttcgatttg atggcgtaaa ctttccccca
     1261 aatggtcctg ttatgcagaa gaagactctg aaatgggagg catccactga gaaaatgtat
     1321 ttgcgtgatg gagtgttgac gggcgatatt accatggctc tgctgcttaa aggagatgtc
     1381 cattaccgat gtgacttcag aactacttac aaatctaggc aggagggtgt caagttgcca
     1441 ggatatcact ttgtcgatca ctgcatcagc atattgaggc atgacaaaga ctacaacgag
     1501 gttaagctgt atgagcatgc tgttgcccat tctggattgc cggacaacgt caagtaaacc
     1561 cagctttctt gtacaaagtt ggcattataa gaaagcattg cttatcaatt tgttgcaacg
     1621 aacaggtcac tatcagtcaa aataaaatca ttatttgcca tccagctgat atcccctata
     1681 gtgagtcgta ttacatggtc atagctgttt cctggcagct ctggcccgtg tctcaaaatc
     1741 tctgatgtta cattgcacaa gataaaaata tatcatcatg aacaataaaa ctgtctgctt
     1801 acataaacag taatacaagg ggtgttatga gccatattca acgggaaacg tcgaggccgc
     1861 gattaaattc caacatggat gctgatttat atgggtataa atgggctcgc gataatgtcg
     1921 ggcaatcagg tgcgacaatc tatcgcttgt atgggaagcc cgatgcgcca gagttgtttc
     1981 tgaaacatgg caaaggtagc gttgccaatg atgttacaga tgagatggtc agactaaact
     2041 ggctgacgga atttatgcct cttccgacca tcaagcattt tatccgtact cctgatgatg
     2101 catggttact caccactgcg atccccggaa aaacagcatt ccaggtatta gaagaatatc
     2161 ctgattcagg tgaaaatatt gttgatgcgc tggcagtgtt cctgcgccgg ttgcattcga
     2221 ttcctgtttg taattgtcct tttaacagcg atcgcgtatt tcgtctcgct caggcgcaat
     2281 cacgaatgaa taacggtttg gttgatgcga gtgattttga tgacgagcgt aatggctggc
     2341 ctgttgaaca agtctggaaa gaaatgcata aacttttgcc attctcaccg gattcagtcg
     2401 tcactcatgg tgatttctca cttgataacc ttatttttga cgaggggaaa ttaataggtt
     2461 gtattgatgt tggacgagtc ggaatcgcag accgatacca ggatcttgcc atcctatgga
     2521 actgcctcgg tgagttttct ccttcattac agaaacggct ttttcaaaaa tatggtattg
     2581 ataatcctga tatgaataaa ttgcagtttc atttgatgct cgatgagttt ttctaatcag
     2641 aattggttaa ttggttgtaa cactggcaga gcattacgct gacttgacgg gacggcgcaa
     2701 gctcatgacc aaaatccctt aacgtgagtt acgcgtcgtt ccactgagcg tcagaccccg
     2761 tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc
     2821 aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc
     2881 tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt
     2941 agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc
     3001 taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact
     3061 caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac
     3121 agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag
     3181 aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtc
//
