LOCUS       sequence-161151-        4206 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Middle entry vector containing GAL4 fused to ERT2 and the VP16 
            transcriptional activation domain for tamoxifen-inducible 
            UAS-promoted transgene expression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4206)
  AUTHORS   Akerberg AA, Stewart S, Stankunas K
  TITLE     Spatial and temporal control of transgene expression in zebrafish.
  JOURNAL   PLoS One. 2014 Mar 18;9(3):e92217. doi: 
            10.1371/journal.pone.0092217. eCollection 2014.
  PUBMED    24643048
REFERENCE   2  (bases 1 to 4206)
  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: "PLoS One.";
            date: "2014-03-18"; volume: "9(3):e92217. doi"; pages: " 
            10.1371/journal.pone.0092217. eCollection 2014"
COMMENT     Sequence Label: pME-GAL4-ERT2-VP16
FEATURES             Location/Qualifiers
     source          1..4206
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     92..109
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(268..295)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(387..473)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     522..544
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     537..553
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    569..668
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     670..686
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        696..714
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     739..755
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     740..756
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2231..2247)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2231..2247)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     primer_bind     complement(2284..2304)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(2284..2302)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     protein_bind    complement(2331..2430)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        complement(2448..2466)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2471..2487)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2471..2487)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2589..2608)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2600..3409
                     /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(2667..2686)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3556..4144
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4045..4064
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga
       61 taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga
      121 gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctggca
      181 cgacaggttt cccgactgga aagcgggcag tgagcgcaac gcaattaata cgcgtaccgc
      241 tagccaggaa gagtttgtag aaacgcaaaa aggccatccg tcaggatggc cttctgctta
      301 gtttgatgcc tggcagttta tggcgggcgt cctgcccgcc accctccggg ccgttgcttc
      361 acaacgttca aatccgctcc cggcggattt gtcctactca ggagagcgtt caccgacaaa
      421 caacagataa aacgaaaggc ccagtcttcc gactgagcct ttcgttttat ttgatgcctg
      481 gcagttccct actctcgcgt taacgctagc atggatgttt tcccagtcac gacgttgtaa
      541 aacgacggcc agtcttaagc tcgggcccca aataatgatt ttattttgac tgatagtgac
      601 ctgttcgttg caacacattg atgagcaatg cttttttata atgccaactt tgtacaaaaa
      661 agcaggcttg taaaacgacg gccagtgagc gcgcgtaata cgactcacta tagggcgaat
      721 tgggtaccgg gccccccctc gaggtcgacg gtatcgataa gcttgatatc gaattcggct
      781 tggggtcgac atgaagctac tgtcttctat cgaacaagca tgcgatattt gccgacttaa
      841 aaagctcaag tgctccaaag aaaaaccgaa gtgcgccaag tgtctgaaga acaactggga
      901 gtgtcgctac tctcccaaaa ccaaaaggtc tccgctgact agggcacatc tgacagaagt
      961 ggaatcaagg ctagaaagac tggaacagct atttctactg atttttcctc gagaagacct
     1021 tgacatgatt ttgaaaatgg attctttaca ggatataaaa gcattgttgg gtacccctgc
     1081 agctgcgtcg actctagagg atccatctgc tggagacatg agagctgcca acctttggcc
     1141 aagcccgctc atgatcaaac gctctaagaa gaacagcctg gccttgtccc tgacggccga
     1201 ccagatggtc agtgccttgt tggatgctga gccccccata ctctattccg agtatgatcc
     1261 taccagaccc ttcagtgaag cttcgatgat gggcttactg accaacctgg cagacaggga
     1321 gctggttcac atgatcaact gggcgaagag ggtgccaggc tttgtggatt tgaccctcca
     1381 tgatcaggtc caccttctag aatgtgcctg gctagagatc ctgatgattg gtctcgtctg
     1441 gcgctccatg gagcacccag tgaagctact gtttgctcct aacttgctct tggacaggaa
     1501 ccagggaaaa tgtgtagagg gcatggtgga gatcttcgac atgctgctgg ctacatcatc
     1561 tcggttccgc atgatgaatc tgcagggaga ggagtttgtg tgcctcaaat ctattatttt
     1621 gcttaattct ggagtgtaca catttctgtc cagcaccctg aagtctctgg aagagaagga
     1681 ccatatccac cgagtcctgg acaagatcac agacactttg atccacctga tggccaaggc
     1741 aggcctgacc ctgcagcagc agcaccagcg gctggcccag ctcctcctca tcctctccca
     1801 catcaggcac atgagtaaca aaggcatgga gcatctgtac agcatgaagt gcaagaacgt
     1861 ggtgcccctc tatgacctgc tgctcgaggc ggcggacgcc caccgcctac atgcgcccac
     1921 tagccgtgga ggggcatccg tggaggagac ggaccaaagc cacttggcca ctgcgggctc
     1981 tacttcatcg gagctccact tagacggcga ggacgtggcg atggcgcatg ccgacgcgct
     2041 agacgatttc gatctggaca tgttggggga cggggattcc ccgggtccgg gatttacccc
     2101 ccacgactcc gccccctacg gcgctctgga tatggccgac ttcgagtttg agcagatgtt
     2161 taccgatgcc cttggaattg acgagtacgg tgggtaagtc gaccccaagc cgaattcctg
     2221 cagcccgggg gatccactag ttctagagcg gccgccaccg cggtggagct ccagcttttg
     2281 ttccctttag tgagggttaa ttgcgcgctt ggcgtaatca tggtcatagc acccagcttt
     2341 cttgtacaaa gttggcatta taagaaagca ttgctcatca atttgttgca acgaacaggt
     2401 cactatcagt caaaataaaa tcattatttg ccatccagct gatatcccct atagtgagtc
     2461 gtattacatg gtcatagctg tttcctggca gctctggccc gtgtctcaaa atctctgatg
     2521 ttacattgca caagataaaa taatatcatc atgaacaata aaactgtctg cttacataaa
     2581 cagtaataca aggggtgtta tgagccatat tcaacgggaa acgtcgaggc cgcgattaaa
     2641 ttccaacatg gatgctgatt tatatgggta taaatgggct cgcgataatg tcgggcaatc
     2701 aggtgcgaca atctatcgct tgtatgggaa gcccgatgcg ccagagttgt ttctgaaaca
     2761 tggcaaaggt agcgttgcca atgatgttac agatgagatg gtcagactaa actggctgac
     2821 ggaatttatg cctcttccga ccatcaagca ttttatccgt actcctgatg atgcatggtt
     2881 actcaccact gcgatccccg gaaaaacagc attccaggta ttagaagaat atcctgattc
     2941 aggtgaaaat attgttgatg cgctggcagt gttcctgcgc cggttgcatt cgattcctgt
     3001 ttgtaattgt ccttttaaca gcgatcgcgt atttcgtctc gctcaggcgc aatcacgaat
     3061 gaataacggt ttggttgatg cgagtgattt tgatgacgag cgtaatggct ggcctgttga
     3121 acaagtctgg aaagaaatgc ataaactttt gccattctca ccggattcag tcgtcactca
     3181 tggtgatttc tcacttgata accttatttt tgacgagggg aaattaatag gttgtattga
     3241 tgttggacga gtcggaatcg cagaccgata ccaggatctt gccatcctat ggaactgcct
     3301 cggtgagttt tctccttcat tacagaaacg gctttttcaa aaatatggta ttgataatcc
     3361 tgatatgaat aaattgcagt ttcatttgat gctcgatgag tttttctaat cagaattggt
     3421 taattggttg taacactggc agagcattac gctgacttga cgggacggcg caagctcatg
     3481 accaaaatcc cttaacgtga gttacgcgtc gttccactga gcgtcagacc ccgtagaaaa
     3541 gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa
     3601 aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc
     3661 gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta
     3721 gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct
     3781 gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg
     3841 atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag
     3901 cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc
     3961 cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg
     4021 agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt
     4081 tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg
     4141 gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca
     4201 catgtt
//
