LOCUS       sequence-385184-        3321 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 3321)
  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 3321)
  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: 768 pME mApple-CAAX
FEATURES             Location/Qualifiers
     source          1..3321
                     /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             138..845
                     /codon_start=1
                     /product="photostable monomeric derivative of DsRed (Shaner
                     et al., 2008)"
                     /label=mApple
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEENNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEA
                     FQTAKLKVTKGGPLPFAWDILSPQFMYGSKVYIKHPADIPDYFKLSFPEGFRWERVMNF
                     EDGGIIHVNQDSSLQDGVFIYKVKLRGTNFPSDGPVMQKKTMGWEASEERMYPEDGALK
                     SEIKKRLKLKDGGHYAAEVKTTYKAKKPVQLPGAYIVDIKLDIVSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     551..570
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     protein_bind    complement(910..1009)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1026..1045)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1027..1045)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1050..1066)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1050..1066)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1168..1187)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             1179..1988
                     /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(1246..1265)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      2135..2723
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2624..2643
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2877..2894
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(3053..3080)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(3172..3258)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     join(3307..3321,1..8)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     join(3321,1..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 gtgagcaagg gcgaggagaa taacatggcc atcatcaagg
      181 agttcatgcg cttcaaggtg cacatggagg gctccgtgaa cggccacgag ttcgagatcg
      241 agggcgaggg cgagggccgc ccctacgagg cctttcagac cgctaagctg aaggtgacca
      301 agggtggccc cctgcccttc gcctgggaca tcctgtcccc tcagttcatg tacggctcca
      361 aggtctacat taagcaccca gccgacatcc ccgactactt caagctgtcc ttccccgagg
      421 gcttcaggtg ggagcgcgtg atgaacttcg aggacggcgg cattattcac gttaaccagg
      481 actcctccct gcaggacggc gtgttcatct acaaggtgaa gctgcgcggc accaacttcc
      541 cctccgacgg ccccgtaatg cagaagaaga ccatgggctg ggaggcctcc gaggagcgga
      601 tgtaccccga ggacggcgcc ctgaagagcg agatcaagaa gaggctgaag ctgaaggacg
      661 gcggccacta cgccgccgag gtcaagacca cctacaaggc caagaagccc gtgcagctgc
      721 ccggcgccta catcgtcgac atcaagttgg acatcgtgtc ccacaacgag gactacacca
      781 tcgtggaaca gtacgaacgc gccgagggcc gccactccac cggcggcatg gacgagctgt
      841 acaagaagct gaaccctcct gatgagagtg gccccggctg catgagctgc aagtgtgtgc
      901 tctcctgaca cccagctttc ttgtacaaag ttggcattat aagaaagcat tgcttatcaa
      961 tttgttgcaa cgaacaggtc actatcagtc aaaataaaat cattatttgc catccagctg
     1021 atatccccta tagtgagtcg tattacatgg tcatagctgt ttcctggcag ctctggcccg
     1081 tgtctcaaaa tctctgatgt tacattgcac aagataaaaa tatatcatca tgaacaataa
     1141 aactgtctgc ttacataaac agtaatacaa ggggtgttat gagccatatt caacgggaaa
     1201 cgtcgaggcc gcgattaaat tccaacatgg atgctgattt atatgggtat aaatgggctc
     1261 gcgataatgt cgggcaatca ggtgcgacaa tctatcgctt gtatgggaag cccgatgcgc
     1321 cagagttgtt tctgaaacat ggcaaaggta gcgttgccaa tgatgttaca gatgagatgg
     1381 tcagactaaa ctggctgacg gaatttatgc ctcttccgac catcaagcat tttatccgta
     1441 ctcctgatga tgcatggtta ctcaccactg cgatccccgg aaaaacagca ttccaggtat
     1501 tagaagaata tcctgattca ggtgaaaata ttgttgatgc gctggcagtg ttcctgcgcc
     1561 ggttgcattc gattcctgtt tgtaattgtc cttttaacag cgatcgcgta tttcgtctcg
     1621 ctcaggcgca atcacgaatg aataacggtt tggttgatgc gagtgatttt gatgacgagc
     1681 gtaatggctg gcctgttgaa caagtctgga aagaaatgca taaacttttg ccattctcac
     1741 cggattcagt cgtcactcat ggtgatttct cacttgataa ccttattttt gacgagggga
     1801 aattaatagg ttgtattgat gttggacgag tcggaatcgc agaccgatac caggatcttg
     1861 ccatcctatg gaactgcctc ggtgagtttt ctccttcatt acagaaacgg ctttttcaaa
     1921 aatatggtat tgataatcct gatatgaata aattgcagtt tcatttgatg ctcgatgagt
     1981 ttttctaatc agaattggtt aattggttgt aacactggca gagcattacg ctgacttgac
     2041 gggacggcgc aagctcatga ccaaaatccc ttaacgtgag ttacgcgtcg ttccactgag
     2101 cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa
     2161 tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag
     2221 agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg
     2281 ttcttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat
     2341 acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta
     2401 ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg
     2461 gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc
     2521 gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa
     2581 gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc
     2641 tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt
     2701 caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct
     2761 tttgctggcc ttttgctcac atgttctttc ctgcgttatc ccctgattct gtggataacc
     2821 gtattaccgc ctttgagtga gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg
     2881 agtcagtgag cgaggaagcg gaagagcgcc caatacgcaa accgcctctc cccgcgcgtt
     2941 ggccgattca ttaatgcagc tggcacgaca ggtttcccga ctggaaagcg ggcagtgagc
     3001 gcaacgcaat taatacgcgt accgctagcc aggaagagtt tgtagaaacg caaaaaggcc
     3061 atccgtcagg atggccttct gcttagtttg atgcctggca gtttatggcg ggcgtcctgc
     3121 ccgccaccct ccgggccgtt gcttcacaac gttcaaatcc gctcccggcg gatttgtcct
     3181 actcaggaga gcgttcaccg acaaacaaca gataaaacga aaggcccagt cttccgactg
     3241 agcctttcgt tttatttgat gcctggcagt tccctactct cgcgttaacg ctagcatgga
     3301 tgttttccca gtcacgacgt t
//
