LOCUS       sequence-8637-ex        2621 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Gateway entry vector for Firefly Luciferase shRNA under H1/TO 
            promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2621)
  AUTHORS   Campeau E, Ruhl VE, Rodier F, Smith CL, Rahmberg BL, Fuss JO, 
            Campisi J, Yaswen P, Cooper PK, Kaufman PD.
  TITLE     A versatile viral system for expression and depletion of proteins in
            mammalian cells.
  JOURNAL   PLoS One. 2009 Aug 6;4(8):e6529.
  PUBMED    19657394
REFERENCE   2  (bases 1 to 2621)
  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: "2009-08-6"; volume: "4(8)"; pages: "e6529"
COMMENT     Sequence Label: pENTR/pTER shLUC (w177-1)
FEATURES             Location/Qualifiers
     source          1..2621
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      103..189
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      281..308
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     primer_bind     316..339
                     /label=pENTR-F
                     /note="pENTR vectors, forward primer"
     protein_bind    358..457
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     634..653
                     /label=H1
                     /note="Human H1 promoter, forward primer"
     protein_bind    699..717
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    complement(850..949)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1061..1080)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             1072..1881
                     /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
                     FQVLEEYPDSGENIVDALAVSLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
                     FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
                     RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     complement(1139..1158)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      1971..2559
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2460..2479
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgctagca tggatctcgg
       61 ggacgtctaa ctactaagcg agagtaggga actgccaggc atcaaataaa acgaaaggct
      121 cagtcggaag actgggcctt tcgttttatc tgttgtttgt cggtgaacgc tctcctgagt
      181 aggacaaatc cgccgggagc ggatttgaac gttgtgaagc aacggcccgg agggtggcgg
      241 gcaggacgcc cgccataaac tgccaggcat caaactaagc agaaggccat cctgacggat
      301 ggcctttttg cgtttctaca aactcttcct gttagttagt tacttaagct cgggccccaa
      361 ataatgattt tattttgact gatagtgacc tgttcgttgc aacaaattga taagcaatgc
      421 ttttttataa tgccaacttt gtacaaaaaa gcaggcttta aaggaaccaa ttcagtcgac
      481 tggatccggt accgaattcg aacgctgacg tcatcaaccc gctccaagga atcgcggccc
      541 agtgtcacta ggcgggaaca cccagcgcgc gtgcgccctg gcaggaagat ggctgtgagg
      601 gacaggggag tggcgccctg caatatttgc atgtcgctat gtgttctggg aaatcaccat
      661 aaacgtgaaa tgtctttgga tttgggaatc ttataagttc cctatcagtg atagagatcc
      721 ccttacgctg agtacttcga gtgtgctgtc ctcgaagtac tcagcgtaag tttttggaaa
      781 agcttggtac cgagctcgga tccactagtc cagtgtggtg gaattcgcgg ccgcactcga
      841 gatatctaga cccagctttc ttgtacaaag ttggcattat aagaaagcat tgcttatcaa
      901 tttgttgcaa cgaacaggtc actatcagtc aaaataaaat cattatttgc catccagctg
      961 cagctctggc ccgtgtctca aaatctctga tgttacattg cacaagataa aaatatatca
     1021 tcatgaacaa taaaactgtc tgcttacata aacagtaata caaggggtgt tatgagccat
     1081 attcaacggg aaacgtcgag gccgcgatta aattccaaca tggatgctga tttatatggg
     1141 tataaatggg ctcgcgataa tgtcgggcaa tcaggtgcga caatctatcg cttgtatggg
     1201 aagcccgatg cgccagagtt gtttctgaaa catggcaaag gtagcgttgc caatgatgtt
     1261 acagatgaga tggtcagact aaactggctg acggaattta tgcctcttcc gaccatcaag
     1321 cattttatcc gtactcctga tgatgcatgg ttactcacca ctgcgatccc cggaaaaaca
     1381 gcattccagg tattagaaga atatcctgat tcaggtgaaa atattgttga tgcgctggca
     1441 gtgtccctgc gccggttgca ttcgattcct gtttgtaatt gtccttttaa cagcgatcgc
     1501 gtatttcgtc tcgctcaggc gcaatcacga atgaataacg gtttggttga tgcgagtgat
     1561 tttgatgacg agcgtaatgg ctggcctgtt gaacaagtct ggaaagaaat gcataaactt
     1621 ttgccattct caccggattc agtcgtcact catggtgatt tctcacttga taaccttatt
     1681 tttgacgagg ggaaattaat aggttgtatt gatgttggac gagtcggaat cgcagaccga
     1741 taccaggatc ttgccatcct atggaactgc ctcggtgagt tttctccttc attacagaaa
     1801 cggctttttc aaaaatatgg tattgataat cctgatatga ataaattgca gtttcatttg
     1861 atgctcgatg agtttttcta atcagaattg gttaattggt tgtaacatta ttcagattgg
     1921 gccccgttcc actgagcgtc agacccggta gaaaagatca aaggatcttc ttgagatcct
     1981 ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt
     2041 tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg
     2101 cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt caagaactct
     2161 gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc
     2221 gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg
     2281 tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa
     2341 ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg
     2401 gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg
     2461 ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga
     2521 tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt
     2581 ttacggttcc tggccttttg ctggcctttt gctcacatgt t
//
