LOCUS       sequence-310836-        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"
     terminator      complement(59..145)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     194..216
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     208..225
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     209..225
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    241..340
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     regulatory      357..366
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1020..1364
                     /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(1442..1461)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1496..1515
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(1578..1594)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1579..1595)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     protein_bind    complement(1621..1720)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        complement(1738..1756)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1761..1777)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1761..1777)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1879..1898)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             1890..2699
                     /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(1957..1976)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      2846..3434
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3335..3354
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3588..3605
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(3764..3791)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
ORIGIN
        1 gtcctgcccg ccaccctccg ggccgttgct tcacaacgtt caaatccgct cccggcggat
       61 ttgtcctact caggagagcg ttcaccgaca aacaacagat aaaacgaaag gcccagtctt
      121 ccgactgagc ctttcgtttt atttgatgcc tggcagttcc ctactctcgc gttaacgcta
      181 gcatggatgt tttcccagtc acgacgttgt aaaacgacgg ccagtcttaa gctcgggccc
      241 caaataatga ttttattttg actgatagtg acctgttcgt tgcaacaaat tgatgagcaa
      301 tgctttttta taatgccaac tttgtacaaa aaagcaggct cgaattggag ctcgccgcca
      361 ccatggcacc caagaagaag aggaagatgc cgcctaaacg caagacactc aatgccgctg
      421 ccgaagccaa tgcccacgct gatggccatg ctgatggcaa tgctgatggt catgtcgcta
      481 acactgcagc aagcagcaac aacgcccgtt ttgcggactt gaccaacatt gacacacccg
      541 gcctcggccc taccacgacg acgttacttg tcgagcccgc tcgttcgaaa cgccagagag
      601 tctcgagggc ctgtgatcag tgtcgagctg cacgtgaaaa gtgtgatgga atccagccgg
      661 cttgcttccc ctgtgtgtcg cagggccggt cgtgtaccta ccaggccagt cccaagaagc
      721 gaggagtcca gacgggctac atccgcactc tcgaactggc tctggcttgg atgttcgaga
      781 acgttgcccg cagcgaggac gccctccaca atcttttggt ccgtgatgct ggccagggca
      841 gcgctctcct ggtcggcaaa gactcgcctg ctgcagaacg cctgcatgca agatgggcga
      901 cgagtcgagt caacaaaagc atcacccgtc ttctctcagg tgctagcact agtagcgccg
      961 gtcaacagca acagctgcaa tcattagtgc cactgacccc gtctgctttg tttggtggcg
     1021 ccaattttaa tcaaagtggg aatattgctg atagctcatt gtccttcact ttcactaaca
     1081 gtagcaacgg tccgaacctc ataacaactc aaacaaattc tcaagcgctt tcacaaccaa
     1141 ttgcctcctc taacgttcat gataacttca tgaataatga aatcacggct agtaaaattg
     1201 atgatggtaa taattcaaaa ccactgtcac ctggttggac ggaccaaact gcgtataacg
     1261 cgtttggaat cactacaggg atgtttaata ccactacaat ggatgatgta tataactatc
     1321 tattcgatga tgaagatacc ccaccaaacc caaaaaaaga gtaacccggg agcggcccct
     1381 ctcgagcctc tagaactata gtgagtcgta ttacgtagat ccagacatga taagatacat
     1441 tgatgagttt ggacaaacca caactagaat gcagtgaaaa aaatgcttta tttgtgaaat
     1501 ttgtgatgct attgctttat ttgtaaccat tataagctgc aataaacaag agatctgatc
     1561 tagaggatca agcttatcga taccgtcgac ctcgaggggg ggcccggtac ccagcttttg
     1621 acccagcttt cttgtacaaa gttggcatta taagaaagca ttgcttatca atttgttgca
     1681 acgaacaggt cactatcagt caaaataaaa tcattatttg ccatccagct gatatcccct
     1741 atagtgagtc gtattacatg gtcatagctg tttcctggca gctctggccc gtgtctcaaa
     1801 atctctgatg ttacattgca caagataaaa atatatcatc atgaacaata aaactgtctg
     1861 cttacataaa cagtaataca aggggtgtta tgagccatat tcaacgggaa acgtcgaggc
     1921 cgcgattaaa ttccaacatg gatgctgatt tatatgggta taaatgggct cgcgataatg
     1981 tcgggcaatc aggtgcgaca atctatcgct tgtatgggaa gcccgatgcg ccagagttgt
     2041 ttctgaaaca tggcaaaggt agcgttgcca atgatgttac agatgagatg gtcagactaa
     2101 actggctgac ggaatttatg cctcttccga ccatcaagca ttttatccgt actcctgatg
     2161 atgcatggtt actcaccact gcgatccccg gaaaaacagc attccaggta ttagaagaat
     2221 atcctgattc aggtgaaaat attgttgatg cgctggcagt gttcctgcgc cggttgcatt
     2281 cgattcctgt ttgtaattgt ccttttaaca gcgatcgcgt atttcgtctc gctcaggcgc
     2341 aatcacgaat gaataacggt ttggttgatg cgagtgattt tgatgacgag cgtaatggct
     2401 ggcctgttga acaagtctgg aaagaaatgc ataaactttt gccattctca ccggattcag
     2461 tcgtcactca tggtgatttc tcacttgata accttatttt tgacgagggg aaattaatag
     2521 gttgtattga tgttggacga gtcggaatcg cagaccgata ccaggatctt gccatcctat
     2581 ggaactgcct cggtgagttt tctccttcat tacagaaacg gctttttcaa aaatatggta
     2641 ttgataatcc tgatatgaat aaattgcagt ttcatttgat gctcgatgag tttttctaat
     2701 cagaattggt taattggttg taacactggc agagcattac gctgacttga cgggacggcg
     2761 caagctcatg accaaaatcc cttaacgtga gttacgcgtc gttccactga gcgtcagacc
     2821 ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct
     2881 tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa
     2941 ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag
     3001 tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc
     3061 tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg
     3121 actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca
     3181 cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat
     3241 gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg
     3301 tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc
     3361 ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc
     3421 ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc
     3481 cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg
     3541 cctttgagtg agctgatacc gctcgccgca gccgaacgac cgagcgcagc gagtcagtga
     3601 gcgaggaagc ggaagagcgc ccaatacgca aaccgcctct ccccgcgcgt tggccgattc
     3661 attaatgcag ctggcacgac aggtttcccg actggaaagc gggcagtgag cgcaacgcaa
     3721 ttaatacgcg taccgctagc caggaagagt ttgtagaaac gcaaaaaggc catccgtcag
     3781 gatggccttc tgcttagttt gatgcctggc agtttatggc gggc
//
