LOCUS       sequence-8636-ex        2620 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Gateway entry vector for EGFPshRNA#2 under H1/TO promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2620)
  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 2620)
  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 shEGFP #2 (728-2)
FEATURES             Location/Qualifiers
     source          1..2620
                     /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(849..948)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1060..1079)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             1071..1880
                     /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(1138..1157)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      1970..2558
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2459..2478
                     /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 ccgctggagt acaactacaa cttcaagaga gttgtagttg tactccagct ttttggaaaa
      781 gcttggtacc gagctcggat ccactagtcc agtgtggtgg aattcgcggc cgcactcgag
      841 atatctagac ccagctttct tgtacaaagt tggcattata agaaagcatt gcttatcaat
      901 ttgttgcaac gaacaggtca ctatcagtca aaataaaatc attatttgcc atccagctgc
      961 agctctggcc cgtgtctcaa aatctctgat gttacattgc acaagataaa aatatatcat
     1021 catgaacaat aaaactgtct gcttacataa acagtaatac aaggggtgtt atgagccata
     1081 ttcaacggga aacgtcgagg ccgcgattaa attccaacat ggatgctgat ttatatgggt
     1141 ataaatgggc tcgcgataat gtcgggcaat caggtgcgac aatctatcgc ttgtatggga
     1201 agcccgatgc gccagagttg tttctgaaac atggcaaagg tagcgttgcc aatgatgtta
     1261 cagatgagat ggtcagacta aactggctga cggaatttat gcctcttccg accatcaagc
     1321 attttatccg tactcctgat gatgcatggt tactcaccac tgcgatcccc ggaaaaacag
     1381 cattccaggt attagaagaa tatcctgatt caggtgaaaa tattgttgat gcgctggcag
     1441 tgtccctgcg ccggttgcat tcgattcctg tttgtaattg tccttttaac agcgatcgcg
     1501 tatttcgtct cgctcaggcg caatcacgaa tgaataacgg tttggttgat gcgagtgatt
     1561 ttgatgacga gcgtaatggc tggcctgttg aacaagtctg gaaagaaatg cataaacttt
     1621 tgccattctc accggattca gtcgtcactc atggtgattt ctcacttgat aaccttattt
     1681 ttgacgaggg gaaattaata ggttgtattg atgttggacg agtcggaatc gcagaccgat
     1741 accaggatct tgccatccta tggaactgcc tcggtgagtt ttctccttca ttacagaaac
     1801 ggctttttca aaaatatggt attgataatc ctgatatgaa taaattgcag tttcatttga
     1861 tgctcgatga gtttttctaa tcagaattgg ttaattggtt gtaacattat tcagattggg
     1921 ccccgttcca ctgagcgtca gacccggtag aaaagatcaa aggatcttct tgagatcctt
     1981 tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt
     2041 gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc
     2101 agataccaaa tactgttctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg
     2161 tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg
     2221 ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt
     2281 cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac
     2341 tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg
     2401 acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg
     2461 gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat
     2521 ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt
     2581 tacggttcct ggccttttgc tggccttttg ctcacatgtt
//
