LOCUS       sequence-263465-        3618 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  full-length of nuclear localized sfGFP with stop codon in pmE 
            Gateway vector.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3618)
  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 3618)
  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-nls-sfGFP
FEATURES             Location/Qualifiers
     source          1..3618
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      complement(85..112)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(204..290)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     339..361
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     353..370
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     354..370
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    386..485
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     regulatory      488..497
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             500..520
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             548..577
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             587..616
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             626..655
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             665..694
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             704..733
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             767..796
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             1460..1507
                     /codon_start=1
                     /product="11th beta-strand of superfolder GFP"
                     /label=GFP11
                     /note="associates with GFP(1-10), consisting of the first 
                     10 beta-strands, to reconstitute fluorescent GFP (Cabantous
                     et al., 2005; Kamiyama et al., 2016)"
                     /translation="RDHMVLHEYVNAAGIT"
     protein_bind    complement(1560..1659)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1676..1695)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1677..1695)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1700..1716)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1700..1716)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1818..1837)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             1829..2638
                     /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(1896..1915)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      2785..3373
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3274..3293
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3527..3544
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 caggtttccc gactggaaag cgggcagtga gcgcaacgca attaatacgc gtaccgctag
       61 ccaggaagag tttgtagaaa cgcaaaaagg ccatccgtca ggatggcctt ctgcttagtt
      121 tgatgcctgg cagtttatgg cgggcgtcct gcccgccacc ctccgggccg ttgcttcaca
      181 acgttcaaat ccgctcccgg cggatttgtc ctactcagga gagcgttcac cgacaaacaa
      241 cagataaaac gaaaggccca gtcttccgac tgagcctttc gttttatttg atgcctggca
      301 gttccctact ctcgcgttaa cgctagcatg gatgttttcc cagtcacgac gttgtaaaac
      361 gacggccagt cttaagctcg ggccccaaat aatgatttta ttttgactga tagtgacctg
      421 ttcgttgcaa caaattgatg agcaatgctt ttttataatg ccaactttgt acaaaaaagc
      481 aggctccgcc accatggctc ctaagaagaa gaggaaggtg gcaggatccc atcgatttaa
      541 agctatggag caaaagctca tttctgaaga ggacttgaat gaaatggagc aaaagctcat
      601 ttctgaagag gacttgaatg aaatggagca aaagctcatt tctgaagagg acttgaatga
      661 aatggagcaa aagctcattt ctgaagagga cttgaatgaa atggagcaaa agctcatttc
      721 tgaagaggac ttgaatgaaa tggagagctt gggcgacctc accatggagc aaaagctcat
      781 ttctgaagag gacttgaatt tagagatctc gaattccatg tccaaaggag aagagctgtt
      841 cactggagtg gtaccaatac ttgtggagtt ggacggagat gtgaacggac acaaattttc
      901 agtccgcggg gagggggaag gggatgctac tattggcaag ctgacgctca aattcatctg
      961 taccaccgga aaactccctg taccctggcc cacactggtg acaactctga cttacggcgt
     1021 gcaatgtttt agccgatacc cagaccacat gaagaggcac gactttttca aaagcgcaat
     1081 gcctgaagga tacgtacagg aaaggaccat ttcttttaaa gacgacggga agtacaaaac
     1141 ccgggcagtg gtgaagtttg agggcgatac cctcgtcaat aggatcgaat tgaagggaac
     1201 tgacttcaaa gaagatggca acatcctggg tcacaagctt gagtataact ttaactccca
     1261 caacgtgtat attacagccg acaaacagaa gaatggaatt aaggctaact tcactgtcag
     1321 acacaatgtc gaagatggct ccgtgcagct cgccgatcac tatcaacaga atactcctat
     1381 cggggacggc ccagtcctgc tgcccgacaa ccactacctg agtacccaga ctgttctgag
     1441 caaagatccg aacgagaagc gcgaccacat ggtgctgcat gagtatgtca acgctgcggg
     1501 aattaccctc ggcatggacg agctgtacaa gtctagagct ccaaagaagc gtaaggtaca
     1561 cccagctttc ttgtacaaag ttggcattat aagaaagcat tgcttatcaa tttgttgcaa
     1621 cgaacaggtc actatcagtc aaaataaaat cattatttgc catccagctg atatccccta
     1681 tagtgagtcg tattacatgg tcatagctgt ttcctggcag ctctggcccg tgtctcaaaa
     1741 tctctgatgt tacattgcac aagataaaaa tatatcatca tgaacaataa aactgtctgc
     1801 ttacataaac agtaatacaa ggggtgttat gagccatatt caacgggaaa cgtcgaggcc
     1861 gcgattaaat tccaacatgg atgctgattt atatgggtat aaatgggctc gcgataatgt
     1921 cgggcaatca ggtgcgacaa tctatcgctt gtatgggaag cccgatgcgc cagagttgtt
     1981 tctgaaacat ggcaaaggta gcgttgccaa tgatgttaca gatgagatgg tcagactaaa
     2041 ctggctgacg gaatttatgc ctcttccgac catcaagcat tttatccgta ctcctgatga
     2101 tgcatggtta ctcaccactg cgatccccgg aaaaacagca ttccaggtat tagaagaata
     2161 tcctgattca ggtgaaaata ttgttgatgc gctggcagtg ttcctgcgcc ggttgcattc
     2221 gattcctgtt tgtaattgtc cttttaacag cgatcgcgta tttcgtctcg ctcaggcgca
     2281 atcacgaatg aataacggtt tggttgatgc gagtgatttt gatgacgagc gtaatggctg
     2341 gcctgttgaa caagtctgga aagaaatgca taaacttttg ccattctcac cggattcagt
     2401 cgtcactcat ggtgatttct cacttgataa ccttattttt gacgagggga aattaatagg
     2461 ttgtattgat gttggacgag tcggaatcgc agaccgatac caggatcttg ccatcctatg
     2521 gaactgcctc ggtgagtttt ctccttcatt acagaaacgg ctttttcaaa aatatggtat
     2581 tgataatcct gatatgaata aattgcagtt tcatttgatg ctcgatgagt ttttctaatc
     2641 agaattggtt aattggttgt aacactggca gagcattacg ctgacttgac gggacggcgc
     2701 aagctcatga ccaaaatccc ttaacgtgag ttacgcgtcg ttccactgag cgtcagaccc
     2761 cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt
     2821 gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac
     2881 tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg ttcttctagt
     2941 gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct
     3001 gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga
     3061 ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac
     3121 acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg
     3181 agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt
     3241 cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc
     3301 tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg
     3361 gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc
     3421 ttttgctcac atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc
     3481 ctttgagtga gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag
     3541 cgaggaagcg gaagagcgcc caatacgcaa accgcctctc cccgcgcgtt ggccgattca
     3601 ttaatgcagc tggcacga
//
