LOCUS       sequence-330918-        2629 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Gateway attB4/attB5 entry clone for inducible mammalian expression 
            using the TRE3G promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2629)
  AUTHORS   Wall VE, Garvey LA, Mehalko JL, Procter LV, Esposito D
  TITLE     Combinatorial assembly of clone libraries using site-specific 
            recombination.
  JOURNAL   Methods Mol Biol. 2014;1116:193-208. doi: 
            10.1007/978-1-62703-764-8_14.
  PUBMED    24395366
REFERENCE   2  (bases 1 to 2629)
  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: "Methods Mol
            Biol."; date: "2014"; volume: "1116:193-208. doi"; pages: " 
            10.1007/978-1-62703-764-8_14"
COMMENT     Sequence Label: C453-E23: attB4-TRE3Gp>-attB5
FEATURES             Location/Qualifiers
     source          1..2629
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    7..102
                     /gene="mutant version of attL"
                     /label=attL5
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        complement(105..483)
                     /label=TRE3G promoter
                     /note="3rd-generation Tet-responsive promoter that can be 
                     activated by binding of Tet-On(R) 3G"
     primer_bind     complement(172..196)
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     protein_bind    241..259
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    277..295
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    313..331
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    349..367
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    385..403
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    421..439
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    457..475
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    complement(483..578)
                     /gene="mutant version of attL"
                     /label=attL4
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     terminator      626..712
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      804..831
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     rep_origin      complement(966..1554)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1046..1065)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(1697..2506)
                     /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     2420..2439
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     2498..2517
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
ORIGIN
        1 agctccaaat aatgatttta ttttgactga tagtgacctg ttcgttgcaa caaattgata
       61 agcaatgctt ttttataatg ccaactttgt atacaaaagt tggctttacg agggtaggaa
      121 gtggtacgga aagttggtat aagacaaaag tgttgtggaa ttgctccagg cgatctgacg
      181 gttcactaaa cgagctctgc ttttataggc gcccaccgta cacgcctaaa gcttatacgt
      241 tctctatcac tgatagggag taaactggat atacgttctc tatcactgat agggagtaaa
      301 ctgtagatac gttctctatc actgataggg agtaaactgg tcatacgttc tctatcactg
      361 atagggagta aactccttat acgttctcta tcactgatag ggagtaaagt ctgcatacgt
      421 tctctatcac tgatagggag taaactcttc atacgttctc tatcactgat agggagtaaa
      481 gccaactttt ctatacaaag ttggcattat aaaaaagcat tgcttatcaa tttgttgcaa
      541 cgaacaggtc actatcagtc aaaataaaat cattatttgg agctccatgg tagcgttaac
      601 gcgagagtag ggaactgcca ggcatcaaat aaaacgaaag gctcagtcgg aagactgggc
      661 ctttcgtttt atctgttgtt tgtcggtgaa cgctctcctg agtaggacaa atccgccggg
      721 agcggatttg aacgttgtga agcaacggcc cggagggtgg cgggcaggac gcccgccata
      781 aactgccagg catcaaacta agcagaaggc catcctgacg gatggccttt ttgcgtttct
      841 acaaactctt cctggctagc ggtaatacgg ttatccacag aatcagggga taacgcagga
      901 aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg
      961 gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag
     1021 aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc
     1081 gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg
     1141 ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt
     1201 cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc
     1261 ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc
     1321 actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg
     1381 tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca
     1441 gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc
     1501 ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat
     1561 cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt
     1621 ttggtcatga gcttgcgccg tcccgtcaag tcagcgtaat gctctgccgg tgttacaacc
     1681 aattaaccaa ttctgattag aaaaactcat cgagcatcaa atgaaactgc aatttattca
     1741 tatcaggatt atcaatacca tatttttgaa aaagccgttt ctgtaatgaa ggagaaaact
     1801 caccgaggca gttccatagg atggcaagat cctggtatcg gtctgcgatt ccgactcgtc
     1861 caacatcaat acaacctatt aatttcccct cgtcaaaaat aaggttatca agtgagaaat
     1921 caccatgagt gacgactgaa tccggcgaga atggcaaaag tttatgcatt tctttccaga
     1981 cttgttcaac aggccagcca ttacgctcgt catcaaaatc actcgcatca accaaaccgt
     2041 tattcattcg tgattgcgcc tgagcgagac gaaatacgcg atcgctgtta aaaggacaat
     2101 tacaaacagg aatcgaatgc aaccggcgca ggaacactgc cagcgcatca acaatatttt
     2161 cacctgaatc aggatattct tctaatacct ggaatgctgt ttttccgggg atcgcagtgg
     2221 tgagtaacca tgcatcatca ggagtacgga taaaatgctt gatggtcgga agaggcataa
     2281 attccgtcag ccagtttagt ctgaccatct catctgtaac atcattggca acgctacctt
     2341 tgccatgttt cagaaacaac tctggcgcat cgggcttccc atacaagcga tagattgtcg
     2401 cacctgattg cccgacatta tcgcgagccc atttataccc atataaatca gcatccatgt
     2461 tggaatttaa tcgcggcctc gacgtttccc gttgaatatg gctcataaca ccccttgtat
     2521 tactgtttat gtaagcagac agttttattg ttcatgatga tatattttta tcttgtgcaa
     2581 tgtaacatca gagattttga gacacgggcc ctgcgccctg cagctctag
//
