LOCUS       sequence-385830-        3182 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Gateway pME Gal4 variant.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3182)
  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 3182)
  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: pCK007 pME KalTa4
FEATURES             Location/Qualifiers
     source          1..3182
                     /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             167..607
                     /codon_start=1
                     /gene="Saccharomyces cerevisiae GAL4 (truncated)"
                     /product="DNA binding domain of the GAL4 transcriptional 
                     activator"
                     /label=GAL4 DNA binding domain
                     /translation="MKLLSSIEQACDICRLKKLKCSKEKPKCAKCLKNNWECRYSPKTK
                     RSPLTRAHLTEVESRLERLEQLFLLIFPREDLDMILKMDSLQDIKALLTGLFVQDNVNK
                     DAVTDRLASVETDMPLTLRQHRISATSSSEESSNKGQRQLTVS"
     protein_bind    complement(771..870)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(887..906)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(888..906)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(911..927)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(911..927)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1029..1048)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             1040..1849
                     /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(1107..1126)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      1996..2584
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2485..2504
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2738..2755
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(2914..2941)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(3033..3119)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     join(3168..3182,1..8)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     join(3182,1..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 gcccatgggc ggccgccccc ttcaccatga aactgctctc
      181 atccatcgag caagcctgcg acatttgtcg gcttaagaag ctgaaatgct ccaaggaaaa
      241 gccgaaatgt gccaaatgcc tgaagaacaa ttgggaatgt cgttactctc ccaaaaccaa
      301 gcgaagtcca ctcacaaggg ctcatctgac cgaagtggag agcaggctag agagactgga
      361 acaactcttt ttgctcatct tccctagaga ggaccttgac atgatcctca agatggattc
      421 tctccaggat attaaagccc ttttgactgg cttattcgtc caggacaatg tgaacaaaga
      481 cgctgtgaca gaccgattgg caagtgtcga gaccgatatg cctctgacac tgagacagca
      541 cagaatcagc gctacttcct caagcgaaga gtcttctaac aagggacaga gacagctgac
      601 tgtttcgagc aggtcgaccc cgtccccggc cgacgccctg gacgacggcg acctggacat
      661 gctgcctgct gatgctctcg atgatttcga tctggatatg ctcccggccg acgccctgga
      721 cgactacgac ctggacatcc tcccgggtaa ctaaaagggt gggcgcgccg acccagcttt
      781 cttgtacaaa gttggcatta taagaaagca ttgcttatca atttgttgca acgaacaggt
      841 cactatcagt caaaataaaa tcattatttg ccatccagct gatatcccct atagtgagtc
      901 gtattacatg gtcatagctg tttcctggca gctctggccc gtgtctcaaa atctctgatg
      961 ttacattgca caagataaaa atatatcatc atgaacaata aaactgtctg cttacataaa
     1021 cagtaataca aggggtgtta tgagccatat tcaacgggaa acgtcgaggc cgcgattaaa
     1081 ttccaacatg gatgctgatt tatatgggta taaatgggct cgcgataatg tcgggcaatc
     1141 aggtgcgaca atctatcgct tgtatgggaa gcccgatgcg ccagagttgt ttctgaaaca
     1201 tggcaaaggt agcgttgcca atgatgttac agatgagatg gtcagactaa actggctgac
     1261 ggaatttatg cctcttccga ccatcaagca ttttatccgt actcctgatg atgcatggtt
     1321 actcaccact gcgatccccg gaaaaacagc attccaggta ttagaagaat atcctgattc
     1381 aggtgaaaat attgttgatg cgctggcagt gttcctgcgc cggttgcatt cgattcctgt
     1441 ttgtaattgt ccttttaaca gcgatcgcgt atttcgtctc gctcaggcgc aatcacgaat
     1501 gaataacggt ttggttgatg cgagtgattt tgatgacgag cgtaatggct ggcctgttga
     1561 acaagtctgg aaagaaatgc ataaactttt gccattctca ccggattcag tcgtcactca
     1621 tggtgatttc tcacttgata accttatttt tgacgagggg aaattaatag gttgtattga
     1681 tgttggacga gtcggaatcg cagaccgata ccaggatctt gccatcctat ggaactgcct
     1741 cggtgagttt tctccttcat tacagaaacg gctttttcaa aaatatggta ttgataatcc
     1801 tgatatgaat aaattgcagt ttcatttgat gctcgatgag tttttctaat cagaattggt
     1861 taattggttg taacactggc agagcattac gctgacttga cgggacggcg caagctcatg
     1921 accaaaatcc cttaacgtga gttacgcgtc gttccactga gcgtcagacc ccgtagaaaa
     1981 gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa
     2041 aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc
     2101 gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta
     2161 gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct
     2221 gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg
     2281 atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag
     2341 cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc
     2401 cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg
     2461 agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt
     2521 tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg
     2581 gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca
     2641 catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg cctttgagtg
     2701 agctgatacc gctcgccgca gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc
     2761 ggaagagcgc ccaatacgca aaccgcctct ccccgcgcgt tggccgattc attaatgcag
     2821 ctggcacgac aggtttcccg actggaaagc gggcagtgag cgcaacgcaa ttaatacgcg
     2881 taccgctagc caggaagagt ttgtagaaac gcaaaaaggc catccgtcag gatggccttc
     2941 tgcttagttt gatgcctggc agtttatggc gggcgtcctg cccgccaccc tccgggccgt
     3001 tgcttcacaa cgttcaaatc cgctcccggc ggatttgtcc tactcaggag agcgttcacc
     3061 gacaaacaac agataaaacg aaaggcccag tcttccgact gagcctttcg ttttatttga
     3121 tgcctggcag ttccctactc tcgcgttaac gctagcatgg atgttttccc agtcacgacg
     3181 tt
//
