LOCUS       sequence-385555-        3437 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Gateway pME fluorophore. *Note: this plasmid has been discontinued. 
            Please see the new, optimized version, Plasmid 213484..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3437)
  AUTHORS   Kemmler CL, Moran HR, Murray BF, Scoresby A, Klem JR, Eckert RL, 
            Lepovsky E, Bertho S, Nieuwenhuize S, Burger S, D'Agati G, Betz C, 
            Puller AC, Felker A, Ditrychova K, Botschi S, Affolter M, Rohner N, 
            Lovely CB, Kwan KM, Burger A, Mosimann C
  TITLE     Next-generation plasmids for transgenesis in zebrafish and beyond.
  JOURNAL   Development. 2023 Mar 28:dev.201531. doi: 10.1242/dev.201531.
  PUBMED    36975217
REFERENCE   2  (bases 1 to 3437)
  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: 
            "Development. 2023 Mar 28:dev.201531. doi: 10.1242/dev.201531."
COMMENT     Sequence Label: pCK006 pME minprom_mCherry no stop
FEATURES             Location/Qualifiers
     source          1..3437
                     /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             296..1003
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYSVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(441..459)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     709..728
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     903..926
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     protein_bind    complement(1026..1125)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1142..1161)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1143..1161)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1166..1182)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1166..1182)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1284..1303)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             1295..2104
                     /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(1362..1381)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      2251..2839
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2740..2759
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2993..3010
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(3169..3196)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(3288..3374)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     join(3423..3437,1..8)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     join(3437,1..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 ctccgcggcc gcccccttag atctcccttc caatctgctc agagaggaca
      181 gagtgggcag gagccagcat tgggtatata aagctgagca gggtcagttg cttcttacgt
      241 ttgcttctga gtctgttgtg ttgacttgca acctcagaaa cagacatccg gatccatggt
      301 gagcaagggc gaggaggaca acatggccat catcaaggag ttcatgcgct tcaaggtgca
      361 catggagggc tccgtgaacg gccacgagtt cgagatcgag ggcgagggcg agggccgccc
      421 ctacgagggc acccagaccg ccaagctgaa ggtgaccaag ggcggccccc tgcccttcgc
      481 ctgggacatc ctgtcccctc agttcatgta cggctccaag gcctacgtga agcaccccgc
      541 cgacatcccc gactacttga agctatcctt ccccgagggc ttcaagtggg agcgcgtgat
      601 gaacttcgag gacggcggcg tggtgaccgt gacccaggac tcctccctgc aggacggcga
      661 gttcatctac aaggtgaagc tgcgcggcac caacttcccc tccgacggcc ccgtaatgca
      721 gaagaagacc atgggctggg aggcctcctc cgagcggatg taccccgagg acggcgccct
      781 gaagggcgag atcaagcaga ggctgaagct gaaggacggc ggccactacg acgccgaggt
      841 caagaccacc tacaaggcca agaagcccgt gcagctgccc ggcgcctaca gcgtcaacat
      901 caagctggac atcacctccc acaacgagga ctacaccatc gtggaacagt acgagcgcgc
      961 cgagggccgc cactccaccg gcggcatgga cgagctgtac aagctcgaga agggtgggcg
     1021 cgccgaccca gctttcttgt acaaagttgg cattataaga aagcattgct tatcaatttg
     1081 ttgcaacgaa caggtcacta tcagtcaaaa taaaatcatt atttgccatc cagctgatat
     1141 cccctatagt gagtcgtatt acatggtcat agctgtttcc tggcagctct ggcccgtgtc
     1201 tcaaaatctc tgatgttaca ttgcacaaga taaaaatata tcatcatgaa caataaaact
     1261 gtctgcttac ataaacagta atacaagggg tgttatgagc catattcaac gggaaacgtc
     1321 gaggccgcga ttaaattcca acatggatgc tgatttatat gggtataaat gggctcgcga
     1381 taatgtcggg caatcaggtg cgacaatcta tcgcttgtat gggaagcccg atgcgccaga
     1441 gttgtttctg aaacatggca aaggtagcgt tgccaatgat gttacagatg agatggtcag
     1501 actaaactgg ctgacggaat ttatgcctct tccgaccatc aagcatttta tccgtactcc
     1561 tgatgatgca tggttactca ccactgcgat ccccggaaaa acagcattcc aggtattaga
     1621 agaatatcct gattcaggtg aaaatattgt tgatgcgctg gcagtgttcc tgcgccggtt
     1681 gcattcgatt cctgtttgta attgtccttt taacagcgat cgcgtatttc gtctcgctca
     1741 ggcgcaatca cgaatgaata acggtttggt tgatgcgagt gattttgatg acgagcgtaa
     1801 tggctggcct gttgaacaag tctggaaaga aatgcataaa cttttgccat tctcaccgga
     1861 ttcagtcgtc actcatggtg atttctcact tgataacctt atttttgacg aggggaaatt
     1921 aataggttgt attgatgttg gacgagtcgg aatcgcagac cgataccagg atcttgccat
     1981 cctatggaac tgcctcggtg agttttctcc ttcattacag aaacggcttt ttcaaaaata
     2041 tggtattgat aatcctgata tgaataaatt gcagtttcat ttgatgctcg atgagttttt
     2101 ctaatcagaa ttggttaatt ggttgtaaca ctggcagagc attacgctga cttgacggga
     2161 cggcgcaagc tcatgaccaa aatcccttaa cgtgagttac gcgtcgttcc actgagcgtc
     2221 agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg
     2281 ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct
     2341 accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct
     2401 tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct
     2461 cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg
     2521 gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc
     2581 gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga
     2641 gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg
     2701 cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta
     2761 tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg
     2821 ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg
     2881 ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat
     2941 taccgccttt gagtgagctg ataccgctcg ccgcagccga acgaccgagc gcagcgagtc
     3001 agtgagcgag gaagcggaag agcgcccaat acgcaaaccg cctctccccg cgcgttggcc
     3061 gattcattaa tgcagctggc acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa
     3121 cgcaattaat acgcgtaccg ctagccagga agagtttgta gaaacgcaaa aaggccatcc
     3181 gtcaggatgg ccttctgctt agtttgatgc ctggcagttt atggcgggcg tcctgcccgc
     3241 caccctccgg gccgttgctt cacaacgttc aaatccgctc ccggcggatt tgtcctactc
     3301 aggagagcgt tcaccgacaa acaacagata aaacgaaagg cccagtcttc cgactgagcc
     3361 tttcgtttta tttgatgcct ggcagttccc tactctcgcg ttaacgctag catggatgtt
     3421 ttcccagtca cgacgtt
//
