LOCUS       sequence-385109-        3288 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 3288)
  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 3288)
  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: 764 pME nls-mApple
FEATURES             Location/Qualifiers
     source          1..3288
                     /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             144..164
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             165..875
                     /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     578..597
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     protein_bind    complement(877..976)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(993..1012)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(994..1012)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1017..1033)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1017..1033)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1135..1154)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             1146..1955
                     /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(1213..1232)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      2102..2690
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2591..2610
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2844..2861
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(3020..3047)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(3139..3225)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     join(3274..3288,1..8)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     join(3288,1..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 gctccaaaga agaagcgtaa ggtaatggtg agcaagggcg
      181 aggagaataa catggccatc atcaaggagt tcatgcgctt caaggtgcac atggagggct
      241 ccgtgaacgg ccacgagttc gagatcgagg gcgagggcga gggccgcccc tacgaggcct
      301 ttcagaccgc taagctgaag gtgaccaagg gtggccccct gcccttcgcc tgggacatcc
      361 tgtcccctca gttcatgtac ggctccaagg tctacattaa gcacccagcc gacatccccg
      421 actacttcaa gctgtccttc cccgagggct tcaggtggga gcgcgtgatg aacttcgagg
      481 acggcggcat tattcacgtt aaccaggact cctccctgca ggacggcgtg ttcatctaca
      541 aggtgaagct gcgcggcacc aacttcccct ccgacggccc cgtaatgcag aagaagacca
      601 tgggctggga ggcctccgag gagcggatgt accccgagga cggcgccctg aagagcgaga
      661 tcaagaagag gctgaagctg aaggacggcg gccactacgc cgccgaggtc aagaccacct
      721 acaaggccaa gaagcccgtg cagctgcccg gcgcctacat cgtcgacatc aagttggaca
      781 tcgtgtccca caacgaggac tacaccatcg tggaacagta cgaacgcgcc gagggccgcc
      841 actccaccgg cggcatggac gagctgtaca agtaacaccc agctttcttg tacaaagttg
      901 gcattataag aaagcattgc ttatcaattt gttgcaacga acaggtcact atcagtcaaa
      961 ataaaatcat tatttgccat ccagctgata tcccctatag tgagtcgtat tacatggtca
     1021 tagctgtttc ctggcagctc tggcccgtgt ctcaaaatct ctgatgttac attgcacaag
     1081 ataaaaatat atcatcatga acaataaaac tgtctgctta cataaacagt aatacaaggg
     1141 gtgttatgag ccatattcaa cgggaaacgt cgaggccgcg attaaattcc aacatggatg
     1201 ctgatttata tgggtataaa tgggctcgcg ataatgtcgg gcaatcaggt gcgacaatct
     1261 atcgcttgta tgggaagccc gatgcgccag agttgtttct gaaacatggc aaaggtagcg
     1321 ttgccaatga tgttacagat gagatggtca gactaaactg gctgacggaa tttatgcctc
     1381 ttccgaccat caagcatttt atccgtactc ctgatgatgc atggttactc accactgcga
     1441 tccccggaaa aacagcattc caggtattag aagaatatcc tgattcaggt gaaaatattg
     1501 ttgatgcgct ggcagtgttc ctgcgccggt tgcattcgat tcctgtttgt aattgtcctt
     1561 ttaacagcga tcgcgtattt cgtctcgctc aggcgcaatc acgaatgaat aacggtttgg
     1621 ttgatgcgag tgattttgat gacgagcgta atggctggcc tgttgaacaa gtctggaaag
     1681 aaatgcataa acttttgcca ttctcaccgg attcagtcgt cactcatggt gatttctcac
     1741 ttgataacct tatttttgac gaggggaaat taataggttg tattgatgtt ggacgagtcg
     1801 gaatcgcaga ccgataccag gatcttgcca tcctatggaa ctgcctcggt gagttttctc
     1861 cttcattaca gaaacggctt tttcaaaaat atggtattga taatcctgat atgaataaat
     1921 tgcagtttca tttgatgctc gatgagtttt tctaatcaga attggttaat tggttgtaac
     1981 actggcagag cattacgctg acttgacggg acggcgcaag ctcatgacca aaatccctta
     2041 acgtgagtta cgcgtcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt
     2101 cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac
     2161 cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct
     2221 tcagcagagc gcagatacca aatactgttc ttctagtgta gccgtagtta ggccaccact
     2281 tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg
     2341 ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata
     2401 aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga
     2461 cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag
     2521 ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg
     2581 agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac
     2641 ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca
     2701 acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg ttctttcctg
     2761 cgttatcccc tgattctgtg gataaccgta ttaccgcctt tgagtgagct gataccgctc
     2821 gccgcagccg aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa gagcgcccaa
     2881 tacgcaaacc gcctctcccc gcgcgttggc cgattcatta atgcagctgg cacgacaggt
     2941 ttcccgactg gaaagcgggc agtgagcgca acgcaattaa tacgcgtacc gctagccagg
     3001 aagagtttgt agaaacgcaa aaaggccatc cgtcaggatg gccttctgct tagtttgatg
     3061 cctggcagtt tatggcgggc gtcctgcccg ccaccctccg ggccgttgct tcacaacgtt
     3121 caaatccgct cccggcggat ttgtcctact caggagagcg ttcaccgaca aacaacagat
     3181 aaaacgaaag gcccagtctt ccgactgagc ctttcgtttt atttgatgcc tggcagttcc
     3241 ctactctcgc gttaacgcta gcatggatgt tttcccagtc acgacgtt
//
