LOCUS       sequence-8621-ex        3313 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Gateway entry vector for shRNA expression under H1/TO promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3313)
  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 3313)
  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+ (430-1)
FEATURES             Location/Qualifiers
     source          1..3313
                     /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(1542..1641)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1753..1772)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             1764..2573
                     /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(1831..1850)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      2663..3251
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3152..3171
                     /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 atagagatct
      721 cggcgttgtg catgtcgggc gactgctcca tgatcttgcg ccgctcgatc tgcgaccaca
      781 ccataaaggc gttcatgggc cgcttgatgt ggccactggg cgtcttgcac cagctggggt
      841 cgtccgcctt gccgcccgtc gacgcggtgg agccaggcgt cggggaggac gcgatgccca
      901 gctccaggcc ggcgcccgag tccgagctct ccccggccag cagagcctca gtgttctccg
      961 cgttgttggt ctgttgtacc atggccccgg cttgccgggc gcccacgcgc gctcacacgc
     1021 tcgcggcgcg gtcgccgggc cctgcgggct ccccggggcg gagaccaggg tcccctgagc
     1081 ggcgcgcggg ccaggggcgg ccgcctcccg ccgccaacct agagtccccg aggccctccg
     1141 tcctccctca ccgctgaagt gctccctgcg ggcaccgagg ctggagtcgg ccgcggcgag
     1201 gagtttctcc aaacgttggt agccggagta tcttccaaac gcaagtttct cgccgcctcc
     1261 ggggcaagtc tttcccccac ccctttccct gaagcagtcg attgcagttc acgagagctc
     1321 gcggtagctc aggaaagcga catcgtctct agcgcctaag tccctttcct gcagtgcaaa
     1381 gccaaagcga ctctggctcc gggactctct gtgggcggaa tcggcgctga ggagttggtg
     1441 caattatttt gttgcaaggt aggaagccaa gaagcttggt accgagctcg gatccactag
     1501 tccagtgtgg tggaattcgc ggccgcactc gagatatcta gacccagctt tcttgtacaa
     1561 agttggcatt ataagaaagc attgcttatc aatttgttgc aacgaacagg tcactatcag
     1621 tcaaaataaa atcattattt gccatccagc tgcagctctg gcccgtgtct caaaatctct
     1681 gatgttacat tgcacaagat aaaaatatat catcatgaac aataaaactg tctgcttaca
     1741 taaacagtaa tacaaggggt gttatgagcc atattcaacg ggaaacgtcg aggccgcgat
     1801 taaattccaa catggatgct gatttatatg ggtataaatg ggctcgcgat aatgtcgggc
     1861 aatcaggtgc gacaatctat cgcttgtatg ggaagcccga tgcgccagag ttgtttctga
     1921 aacatggcaa aggtagcgtt gccaatgatg ttacagatga gatggtcaga ctaaactggc
     1981 tgacggaatt tatgcctctt ccgaccatca agcattttat ccgtactcct gatgatgcat
     2041 ggttactcac cactgcgatc cccggaaaaa cagcattcca ggtattagaa gaatatcctg
     2101 attcaggtga aaatattgtt gatgcgctgg cagtgtccct gcgccggttg cattcgattc
     2161 ctgtttgtaa ttgtcctttt aacagcgatc gcgtatttcg tctcgctcag gcgcaatcac
     2221 gaatgaataa cggtttggtt gatgcgagtg attttgatga cgagcgtaat ggctggcctg
     2281 ttgaacaagt ctggaaagaa atgcataaac ttttgccatt ctcaccggat tcagtcgtca
     2341 ctcatggtga tttctcactt gataacctta tttttgacga ggggaaatta ataggttgta
     2401 ttgatgttgg acgagtcgga atcgcagacc gataccagga tcttgccatc ctatggaact
     2461 gcctcggtga gttttctcct tcattacaga aacggctttt tcaaaaatat ggtattgata
     2521 atcctgatat gaataaattg cagtttcatt tgatgctcga tgagtttttc taatcagaat
     2581 tggttaattg gttgtaacat tattcagatt gggccccgtt ccactgagcg tcagacccgg
     2641 tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc
     2701 aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc
     2761 tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt
     2821 agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc
     2881 taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact
     2941 caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac
     3001 agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag
     3061 aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg
     3121 gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg
     3181 tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga
     3241 gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt
     3301 ttgctcacat gtt
//
