LOCUS       sequence-304792-        3824 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Middle entry vector containing QFGal4.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3824)
  AUTHORS   Burgess J, Burrows JT, Sadhak R, Chiang S, Weiss A, D'Amata C, 
            Molinaro AM, Zhu S, Long M, Hu C, Krause HM, Pearson BJ
  TITLE     An optimized QF-binary expression system for use in zebrafish.
  JOURNAL   Dev Biol. 2020 Jul 19. pii: S0012-1606(20)30202-5. doi: 
            10.1016/j.ydbio.2020.07.007.
  PUBMED    32697972
REFERENCE   2  (bases 1 to 3824)
  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: "Dev Biol. 
            2020 Jul 19. pii: S0012-1606(20)30202-5. doi: 
            10.1016/j.ydbio.2020.07.007."
COMMENT     Sequence Label: pME-QFGal4-SV40pA
FEATURES             Location/Qualifiers
     source          1..3824
                     /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     536..553
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     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"
     regulatory      685..694
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1348..1692
                     /codon_start=1
                     /gene="Saccharomyces cerevisiae GAL4 (truncated)"
                     /product="activation domain of the GAL4 transcriptional 
                     activator"
                     /label=GAL4 activation domain
                     /translation="ANFNQSGNIADSSLSFTFTNSSNGPNLITTQTNSQALSQPIASSN
                     VHDNFMNNEITASKIDDGNNSKPLSPGWTDQTAYNAFGITTGMFNTTTMDDVYNYLFDD
                     EDTPPNPKKE"
     primer_bind     complement(1770..1789)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1824..1843
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(1906..1922)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1907..1923)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     protein_bind    complement(1949..2048)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        complement(2066..2084)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2089..2105)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2089..2105)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2207..2226)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2218..3027
                     /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(2285..2304)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3174..3762
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3663..3682
                     /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 caacaaattg atgagcaatg cttttttata atgccaactt tgtacaaaaa
      661 agcaggctcg aattggagct cgccgccacc atggcaccca agaagaagag gaagatgccg
      721 cctaaacgca agacactcaa tgccgctgcc gaagccaatg cccacgctga tggccatgct
      781 gatggcaatg ctgatggtca tgtcgctaac actgcagcaa gcagcaacaa cgcccgtttt
      841 gcggacttga ccaacattga cacacccggc ctcggcccta ccacgacgac gttacttgtc
      901 gagcccgctc gttcgaaacg ccagagagtc tcgagggcct gtgatcagtg tcgagctgca
      961 cgtgaaaagt gtgatggaat ccagccggct tgcttcccct gtgtgtcgca gggccggtcg
     1021 tgtacctacc aggccagtcc caagaagcga ggagtccaga cgggctacat ccgcactctc
     1081 gaactggctc tggcttggat gttcgagaac gttgcccgca gcgaggacgc cctccacaat
     1141 cttttggtcc gtgatgctgg ccagggcagc gctctcctgg tcggcaaaga ctcgcctgct
     1201 gcagaacgcc tgcatgcaag atgggcgacg agtcgagtca acaaaagcat cacccgtctt
     1261 ctctcaggtg ctagcactag tagcgccggt caacagcaac agctgcaatc attagtgcca
     1321 ctgaccccgt ctgctttgtt tggtggcgcc aattttaatc aaagtgggaa tattgctgat
     1381 agctcattgt ccttcacttt cactaacagt agcaacggtc cgaacctcat aacaactcaa
     1441 acaaattctc aagcgctttc acaaccaatt gcctcctcta acgttcatga taacttcatg
     1501 aataatgaaa tcacggctag taaaattgat gatggtaata attcaaaacc actgtcacct
     1561 ggttggacgg accaaactgc gtataacgcg tttggaatca ctacagggat gtttaatacc
     1621 actacaatgg atgatgtata taactatcta ttcgatgatg aagatacccc accaaaccca
     1681 aaaaaagagt aacccgggag cggcccctct cgagcctcta gaactatagt gagtcgtatt
     1741 acgtagatcc agacatgata agatacattg atgagtttgg acaaaccaca actagaatgc
     1801 agtgaaaaaa atgctttatt tgtgaaattt gtgatgctat tgctttattt gtaaccatta
     1861 taagctgcaa taaacaagag atctgatcta gaggatcaag cttatcgata ccgtcgacct
     1921 cgaggggggg cccggtaccc agcttttgac ccagctttct tgtacaaagt tggcattata
     1981 agaaagcatt gcttatcaat ttgttgcaac gaacaggtca ctatcagtca aaataaaatc
     2041 attatttgcc atccagctga tatcccctat agtgagtcgt attacatggt catagctgtt
     2101 tcctggcagc tctggcccgt gtctcaaaat ctctgatgtt acattgcaca agataaaaat
     2161 atatcatcat gaacaataaa actgtctgct tacataaaca gtaatacaag gggtgttatg
     2221 agccatattc aacgggaaac gtcgaggccg cgattaaatt ccaacatgga tgctgattta
     2281 tatgggtata aatgggctcg cgataatgtc gggcaatcag gtgcgacaat ctatcgcttg
     2341 tatgggaagc ccgatgcgcc agagttgttt ctgaaacatg gcaaaggtag cgttgccaat
     2401 gatgttacag atgagatggt cagactaaac tggctgacgg aatttatgcc tcttccgacc
     2461 atcaagcatt ttatccgtac tcctgatgat gcatggttac tcaccactgc gatccccgga
     2521 aaaacagcat tccaggtatt agaagaatat cctgattcag gtgaaaatat tgttgatgcg
     2581 ctggcagtgt tcctgcgccg gttgcattcg attcctgttt gtaattgtcc ttttaacagc
     2641 gatcgcgtat ttcgtctcgc tcaggcgcaa tcacgaatga ataacggttt ggttgatgcg
     2701 agtgattttg atgacgagcg taatggctgg cctgttgaac aagtctggaa agaaatgcat
     2761 aaacttttgc cattctcacc ggattcagtc gtcactcatg gtgatttctc acttgataac
     2821 cttatttttg acgaggggaa attaataggt tgtattgatg ttggacgagt cggaatcgca
     2881 gaccgatacc aggatcttgc catcctatgg aactgcctcg gtgagttttc tccttcatta
     2941 cagaaacggc tttttcaaaa atatggtatt gataatcctg atatgaataa attgcagttt
     3001 catttgatgc tcgatgagtt tttctaatca gaattggtta attggttgta acactggcag
     3061 agcattacgc tgacttgacg ggacggcgca agctcatgac caaaatccct taacgtgagt
     3121 tacgcgtcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat
     3181 cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg
     3241 gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga
     3301 gcgcagatac caaatactgt tcttctagtg tagccgtagt taggccacca cttcaagaac
     3361 tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt
     3421 ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag
     3481 cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc
     3541 gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag
     3601 gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca
     3661 gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt
     3721 cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc
     3781 tttttacggt tcctggcctt ttgctggcct tttgctcaca tgtt
//
