LOCUS       sequence-385574-        3462 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Gateway pME fluorophore.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3462)
  AUTHORS   Prummel KD, Hess C, Nieuwenhuize S, Parker HJ, Rogers KW, Kozmikova 
            I, Racioppi C, Brombacher EC, Czarkwiani A, Knapp D, Burger S, 
            Chiavacci E, Shah G, Burger A, Huisken J, Yun MH, Christiaen L, 
            Kozmik Z, Muller P, Bronner M, Krumlauf R, Mosimann C
  TITLE     A conserved regulatory program initiates lateral plate mesoderm 
            emergence across chordates.
  JOURNAL   Nat Commun. 2019 Aug 26;10(1):3857. doi: 10.1038/s41467-019-11561-7.
  PUBMED    31451684
REFERENCE   2  (bases 1 to 3462)
  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: "Nat 
            Commun."; date: "2019-08-26"; volume: "10(1):3857. doi"; pages: " 
            10.1038/s41467-019-11561-7"
COMMENT     Sequence Label: pKD001 pME minprom_EGFP
FEATURES             Location/Qualifiers
     source          1..3462
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     1..17
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    33..132
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             join(273..275,276..278,279..989)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(273..275,276..278,279..989)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(318..339)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(579..598)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     926..947
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     protein_bind    complement(1051..1150)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1167..1186)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1168..1186)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1191..1207)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1191..1207)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1309..1328)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             1320..2129
                     /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(1387..1406)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      2276..2864
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2765..2784
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3018..3035
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(3194..3221)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(3313..3399)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     join(3448..3462,1..8)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     join(3462,1..17)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
ORIGIN
        1 gtaaaacgac ggccagtctt aagctcgggc cccaaataat gattttattt tgactgatag
       61 tgacctgttc gttgcaacaa attgatgagc aatgcttttt tataatgcca actttgtaca
      121 aaaaagcagg ctggaccatg gccaatctgc tcagagagga cagagtgggc aggagccagc
      181 attgggtata taaagctgag cagggtcagt tgcttcttac gtttgcttct gagtctgttg
      241 tgttgacttg caacctcaga aacagacatc ccatggtgag caagggcgag gagctgttca
      301 ccggggtggt gcccatcctg gtcgagctgg acggcgacgt aaacggccac aagttcagcg
      361 tgtccggcga gggcgagggc gatgccacct acggcaagct gaccctgaag ttcatctgca
      421 ccaccggcaa gctgcccgtg ccctggccca ccctcgtgac caccctgacc tacggcgtgc
      481 agtgcttcag ccgctacccc gaccacatga agcagcacga cttcttcaag tccgccatgc
      541 ccgaaggcta cgtccaggag cgcaccatct tcttcaagga cgacggcaac tacaagaccc
      601 gcgccgaggt gaagttcgag ggcgacaccc tggtgaaccg catcgagctg aagggcatcg
      661 acttcaagga ggacggcaac atcctggggc acaagctgga gtacaactac aacagccaca
      721 acgtctatat catggccgac aagcagaaga acggcatcaa ggtgaacttc aagatccgcc
      781 acaacatcga ggacggcagc gtgcagctcg ccgaccacta ccagcagaac acccccatcg
      841 gcgacggccc cgtgctgctg cccgacaacc actacctgag cacccagtcc gccctgagca
      901 aagaccccaa cgagaagcgc gatcacatgg tcctgctgga gttcgtgacc gccgccggga
      961 tcactctcgg catggacgag ctgtacaagg gcggtggaag atctgggaat tcaaggcctc
     1021 tcgagcctct agattctgca gccctatagc acccagcttt cttgtacaaa gttggcatta
     1081 taagaaagca ttgcttatca atttgttgca acgaacaggt cactatcagt caaaataaaa
     1141 tcattatttg ccatccagct gatatcccct atagtgagtc gtattacatg gtcatagctg
     1201 tttcctggca gctctggccc gtgtctcaaa atctctgatg ttacattgca caagataaaa
     1261 atatatcatc atgaacaata aaactgtctg cttacataaa cagtaataca aggggtgtta
     1321 tgagccatat tcaacgggaa acgtcgaggc cgcgattaaa ttccaacatg gatgctgatt
     1381 tatatgggta taaatgggct cgcgataatg tcgggcaatc aggtgcgaca atctatcgct
     1441 tgtatgggaa gcccgatgcg ccagagttgt ttctgaaaca tggcaaaggt agcgttgcca
     1501 atgatgttac agatgagatg gtcagactaa actggctgac ggaatttatg cctcttccga
     1561 ccatcaagca ttttatccgt actcctgatg atgcatggtt actcaccact gcgatccccg
     1621 gaaaaacagc attccaggta ttagaagaat atcctgattc aggtgaaaat attgttgatg
     1681 cgctggcagt gttcctgcgc cggttgcatt cgattcctgt ttgtaattgt ccttttaaca
     1741 gcgatcgcgt atttcgtctc gctcaggcgc aatcacgaat gaataacggt ttggttgatg
     1801 cgagtgattt tgatgacgag cgtaatggct ggcctgttga acaagtctgg aaagaaatgc
     1861 ataaactttt gccattctca ccggattcag tcgtcactca tggtgatttc tcacttgata
     1921 accttatttt tgacgagggg aaattaatag gttgtattga tgttggacga gtcggaatcg
     1981 cagaccgata ccaggatctt gccatcctat ggaactgcct cggtgagttt tctccttcat
     2041 tacagaaacg gctttttcaa aaatatggta ttgataatcc tgatatgaat aaattgcagt
     2101 ttcatttgat gctcgatgag tttttctaat cagaattggt taattggttg taacactggc
     2161 agagcattac gctgacttga cgggacggcg caagctcatg accaaaatcc cttaacgtga
     2221 gttacgcgtc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag
     2281 atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg
     2341 tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca
     2401 gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac cacttcaaga
     2461 actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca
     2521 gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc
     2581 agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca
     2641 ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa
     2701 aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc
     2761 cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc
     2821 gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg
     2881 cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt cctgcgttat
     2941 cccctgattc tgtggataac cgtattaccg cctttgagtg agctgatacc gctcgccgca
     3001 gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc ggaagagcgc ccaatacgca
     3061 aaccgcctct ccccgcgcgt tggccgattc attaatgcag ctggcacgac aggtttcccg
     3121 actggaaagc gggcagtgag cgcaacgcaa ttaatacgcg taccgctagc caggaagagt
     3181 ttgtagaaac gcaaaaaggc catccgtcag gatggccttc tgcttagttt gatgcctggc
     3241 agtttatggc gggcgtcctg cccgccaccc tccgggccgt tgcttcacaa cgttcaaatc
     3301 cgctcccggc ggatttgtcc tactcaggag agcgttcacc gacaaacaac agataaaacg
     3361 aaaggcccag tcttccgact gagcctttcg ttttatttga tgcctggcag ttccctactc
     3421 tcgcgttaac gctagcatgg atgttttccc agtcacgacg tt
//
