LOCUS       sequence-8562-ex        3287 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Gateway entry vector for shRNA expression under H1 promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3287)
  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 3287)
  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/pSUPER+ (575-1)
FEATURES             Location/Qualifiers
     source          1..3287
                     /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"
     promoter        509..723
                     /label=H1 promoter
                     /note="human H1 RNA promoter"
     primer_bind     644..663
                     /label=H1
                     /note="Human H1 promoter, forward primer"
     primer_bind     complement(1489..1505)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1490..1506)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     protein_bind    complement(1516..1615)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1727..1746)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             1738..2547
                     /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(1805..1824)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      2637..3225
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3126..3145
                     /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 gcaggctcca ccatgggaac caattcagtc
      481 gactggatcc cccgggctgc aggaattcga acgctgacgt catcaacccg ctccaaggaa
      541 tcgcgggccc agtgtcacta ggcgggaaca cccagcgcgc gtgcgccctg gcaggaagat
      601 ggctgtgagg gacaggggag tggcgccctg caatatttgc atgtcgctat gtgttctggg
      661 aaatcaccat aaacgtgaaa tgtctttgga tttgggaatc ttataagttc tgtatgagac
      721 cacagatctc ggcgttgtgc atgtcgggcg actgctccat gatcttgcgc cgctcgatct
      781 gcgaccacac cataaaggcg ttcatgggcc gcttgatgtg gccactgggc gtcttgcacc
      841 agctggggtc gtccgccttg ccgcccgtcg acgcggtgga gccaggcgtc ggggaggacg
      901 cgatgcccag ctccaggccg gcgcccgagt ccgagctctc cccggccagc agagcctcag
      961 tgttctccgc gttgttggtc tgttgtacca tggccccggc ttgccgggcg cccacgcgcg
     1021 ctcacacgct cgcggcgcgg tcgccgggcc ctgcgggctc cccggggcgg agaccagggt
     1081 cccctgagcg gcgcgcgggc caggggcggc cgcctcccgc cgccaaccta gagtccccga
     1141 ggccctccgt cctccctcac cgctgaagtg ctccctgcgg gcaccgaggc tggagtcggc
     1201 cgcggcgagg agtttctcca aacgttggta gccggagtat cttccaaacg caagtttctc
     1261 gccgcctccg gggcaagtct ttcccccacc cctttccctg aagcagtcga ttgcagttca
     1321 cgagagctcg cggtagctca ggaaagcgac atcgtctcta gcgcctaagt ccctttcctg
     1381 cagtgcaaag ccaaagcgac tctggctccg ggactctctg tgggcggaat cggcgctgag
     1441 gagttggtgc aattattttg ttgcaaggta ggaagccaag aagcttatcg ataccgtcga
     1501 cctcgagata tctagaccca gctttcttgt acaaagttgg cattataaga aagcattgct
     1561 tatcaatttg ttgcaacgaa caggtcacta tcagtcaaaa taaaatcatt atttgccatc
     1621 cagctgcagc tctggcccgt gtctcaaaat ctctgatgtt acattgcaca agataaaaat
     1681 atatcatcat gaacaataaa actgtctgct tacataaaca gtaatacaag gggtgttatg
     1741 agccatattc aacgggaaac gtcgaggccg cgattaaatt ccaacatgga tgctgattta
     1801 tatgggtata aatgggctcg cgataatgtc gggcaatcag gtgcgacaat ctatcgcttg
     1861 tatgggaagc ccgatgcgcc agagttgttt ctgaaacatg gcaaaggtag cgttgccaat
     1921 gatgttacag atgagatggt cagactaaac tggctgacgg aatttatgcc tcttccgacc
     1981 atcaagcatt ttatccgtac tcctgatgat gcatggttac tcaccactgc gatccccgga
     2041 aaaacagcat tccaggtatt agaagaatat cctgattcag gtgaaaatat tgttgatgcg
     2101 ctggcagtgt ccctgcgccg gttgcattcg attcctgttt gtaattgtcc ttttaacagc
     2161 gatcgcgtat ttcgtctcgc tcaggcgcaa tcacgaatga ataacggttt ggttgatgcg
     2221 agtgattttg atgacgagcg taatggctgg cctgttgaac aagtctggaa agaaatgcat
     2281 aaacttttgc cattctcacc ggattcagtc gtcactcatg gtgatttctc acttgataac
     2341 cttatttttg acgaggggaa attaataggt tgtattgatg ttggacgagt cggaatcgca
     2401 gaccgatacc aggatcttgc catcctatgg aactgcctcg gtgagttttc tccttcatta
     2461 cagaaacggc tttttcaaaa atatggtatt gataatcctg atatgaataa attgcagttt
     2521 catttgatgc tcgatgagtt tttctaatca gaattggtta attggttgta acattattca
     2581 gattgggccc cgttccactg agcgtcagac ccggtagaaa agatcaaagg atcttcttga
     2641 gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg
     2701 gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc
     2761 agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca ccacttcaag
     2821 aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc
     2881 agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg
     2941 cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac
     3001 accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga
     3061 aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt
     3121 ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag
     3181 cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg
     3241 gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgtt
//
