LOCUS       sequence-385844-        3317 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 3317)
  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 3317)
  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: pME minprom KalTa4
FEATURES             Location/Qualifiers
     source          1..3317
                     /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             302..742
                     /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(906..1005)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1022..1041)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1023..1041)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1046..1062)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1046..1062)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1164..1183)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             1175..1984
                     /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(1242..1261)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      2131..2719
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2620..2639
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2873..2890
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(3049..3076)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(3168..3254)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     join(3303..3317,1..8)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     join(3317,1..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 gcccatggcc aatctgctca gagaggacag agtgggcagg
      181 agccagcatt gggtatataa agctgagcag ggtcagttgc ttcttacgtt tgcttctgag
      241 tctgttgtgt tgacttgcaa cctcagaaac agacatccca tgggcggccg cccccttcac
      301 catgaaactg ctctcatcca tcgagcaagc ctgcgacatt tgtcggctta agaagctgaa
      361 atgctccaag gaaaagccga aatgtgccaa atgcctgaag aacaattggg aatgtcgtta
      421 ctctcccaaa accaagcgaa gtccactcac aagggctcat ctgaccgaag tggagagcag
      481 gctagagaga ctggaacaac tctttttgct catcttccct agagaggacc ttgacatgat
      541 cctcaagatg gattctctcc aggatattaa agcccttttg actggcttat tcgtccagga
      601 caatgtgaac aaagacgctg tgacagaccg attggcaagt gtcgagaccg atatgcctct
      661 gacactgaga cagcacagaa tcagcgctac ttcctcaagc gaagagtctt ctaacaaggg
      721 acagagacag ctgactgttt cgagcaggtc gaccccgtcc ccggccgacg ccctggacga
      781 cggcgacctg gacatgctgc ctgctgatgc tctcgatgat ttcgatctgg atatgctccc
      841 ggccgacgcc ctggacgact acgacctgga catcctcccg ggtaactaaa agggtgggcg
      901 cgccgaccca gctttcttgt acaaagttgg cattataaga aagcattgct tatcaatttg
      961 ttgcaacgaa caggtcacta tcagtcaaaa taaaatcatt atttgccatc cagctgatat
     1021 cccctatagt gagtcgtatt acatggtcat agctgtttcc tggcagctct ggcccgtgtc
     1081 tcaaaatctc tgatgttaca ttgcacaaga taaaaatata tcatcatgaa caataaaact
     1141 gtctgcttac ataaacagta atacaagggg tgttatgagc catattcaac gggaaacgtc
     1201 gaggccgcga ttaaattcca acatggatgc tgatttatat gggtataaat gggctcgcga
     1261 taatgtcggg caatcaggtg cgacaatcta tcgcttgtat gggaagcccg atgcgccaga
     1321 gttgtttctg aaacatggca aaggtagcgt tgccaatgat gttacagatg agatggtcag
     1381 actaaactgg ctgacggaat ttatgcctct tccgaccatc aagcatttta tccgtactcc
     1441 tgatgatgca tggttactca ccactgcgat ccccggaaaa acagcattcc aggtattaga
     1501 agaatatcct gattcaggtg aaaatattgt tgatgcgctg gcagtgttcc tgcgccggtt
     1561 gcattcgatt cctgtttgta attgtccttt taacagcgat cgcgtatttc gtctcgctca
     1621 ggcgcaatca cgaatgaata acggtttggt tgatgcgagt gattttgatg acgagcgtaa
     1681 tggctggcct gttgaacaag tctggaaaga aatgcataaa cttttgccat tctcaccgga
     1741 ttcagtcgtc actcatggtg atttctcact tgataacctt atttttgacg aggggaaatt
     1801 aataggttgt attgatgttg gacgagtcgg aatcgcagac cgataccagg atcttgccat
     1861 cctatggaac tgcctcggtg agttttctcc ttcattacag aaacggcttt ttcaaaaata
     1921 tggtattgat aatcctgata tgaataaatt gcagtttcat ttgatgctcg atgagttttt
     1981 ctaatcagaa ttggttaatt ggttgtaaca ctggcagagc attacgctga cttgacggga
     2041 cggcgcaagc tcatgaccaa aatcccttaa cgtgagttac gcgtcgttcc actgagcgtc
     2101 agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg
     2161 ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct
     2221 accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct
     2281 tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct
     2341 cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg
     2401 gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc
     2461 gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga
     2521 gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg
     2581 cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta
     2641 tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg
     2701 ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg
     2761 ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat
     2821 taccgccttt gagtgagctg ataccgctcg ccgcagccga acgaccgagc gcagcgagtc
     2881 agtgagcgag gaagcggaag agcgcccaat acgcaaaccg cctctccccg cgcgttggcc
     2941 gattcattaa tgcagctggc acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa
     3001 cgcaattaat acgcgtaccg ctagccagga agagtttgta gaaacgcaaa aaggccatcc
     3061 gtcaggatgg ccttctgctt agtttgatgc ctggcagttt atggcgggcg tcctgcccgc
     3121 caccctccgg gccgttgctt cacaacgttc aaatccgctc ccggcggatt tgtcctactc
     3181 aggagagcgt tcaccgacaa acaacagata aaacgaaagg cccagtcttc cgactgagcc
     3241 tttcgtttta tttgatgcct ggcagttccc tactctcgcg ttaacgctag catggatgtt
     3301 ttcccagtca cgacgtt
//
