LOCUS       sequence-80282-e        2804 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  dual shRNA expression cassette entry vector.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2804)
  AUTHORS   Radulovich N, Leung L, Tsao MS
  TITLE     Modified gateway system for double shRNA expression and Cre/lox 
            based gene expression.
  JOURNAL   BMC Biotechnol. 2011 Mar 22;11:24.
  PUBMED    21418658
REFERENCE   2  (bases 1 to 2804)
  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: "BMC 
            Biotechnol."; date: "2011-03-22"; volume: "11"; pages: "24"
COMMENT     Sequence Label: pENTR.hU6hH1
FEATURES             Location/Qualifiers
     source          1..2804
                     /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        501..749
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     501..521
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     672..691
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     promoter        777..991
                     /label=H1 promoter
                     /note="human H1 RNA promoter"
     primer_bind     912..931
                     /label=H1
                     /note="Human H1 promoter, forward primer"
     primer_bind     complement(1006..1022)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1007..1023)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     protein_bind    complement(1033..1132)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1244..1263)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             1255..2064
                     /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
                     DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPGDAWLLTTAIPGKTA
                     FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
                     FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
                     RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     complement(1322..1341)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      2154..2742
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2643..2662
                     /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 gagcggccgc ccccttcacc gagggcctat ttcccatgat tccttcatat ttgcatatac
      541 gatacaaggc tgttagagag ataattggaa ttaatttgac tgtaaacaca aagatattag
      601 tacaaaatac gtgacgtaga aagtaataat ttcttgggta gtttgcagtt ttaaaattat
      661 gttttaaaat ggactatcat atgcttaccg taacttgaaa gtatttcgat ttcttggctt
      721 tatatatctt gtggaaagga cgaaacaccg gtccgcaggt atgcacgcgt gaattcgaac
      781 gctgacgtca tcaacccgct ccaaggaatc gcgggcccag tgtcactagg cgggaacacc
      841 cagcgcgcgt gcgccctggc aggaagatgg ctgtgaggga caggggagtg gcgccctgca
      901 atatttgcat gtcgctatgt gttctgggaa atcaccataa acgtgaaatg tctttggatt
      961 tgggaatctt ataagttctg tatgagacca cagatctaag cttatcgata ccgtcgacct
     1021 cgagatatct agacccagct ttcttgtaca aagttggcat tataagaaag cattgcttat
     1081 caatttgttg caacgaacag gtcactatca gtcaaaataa aatcattatt tgccatccag
     1141 ctgcagctct ggcccgtgtc tcaaaatctc tgatgttaca ttgcacaaga taaaaatata
     1201 tcatcatgaa caataaaact gtctgcttac ataaacagta atacaagggg tgttatgagc
     1261 catattcaac gggaaacgtc gaggccgcga ttaaattcca acatggatgc tgatttatat
     1321 gggtataaat gggctcgcga taatgtcggg caatcaggtg cgacaatcta tcgcttgtat
     1381 gggaagcccg atgcgccaga gttgtttctg aaacatggca aaggtagcgt tgccaatgat
     1441 gttacagatg agatggtcag actaaactgg ctgacggaat ttatgcctct tccgaccatc
     1501 aagcatttta tccgtactcc tggtgatgca tggttactca ccactgcgat ccccggaaaa
     1561 acagcattcc aggtattaga agaatatcct gattcaggtg aaaatattgt tgatgcgctg
     1621 gcagtgttcc tgcgccggtt gcattcgatt cctgtttgta attgtccttt taacagcgat
     1681 cgcgtatttc gtctcgctca ggcgcaatca cgaatgaata acggtttggt tgatgcgagt
     1741 gattttgatg acgagcgtaa tggctggcct gttgaacaag tctggaaaga aatgcataaa
     1801 cttttgccat tctcaccgga ttcagtcgtc actcatggtg atttctcact tgataacctt
     1861 atttttgacg aggggaaatt aataggttgt attgatgttg gacgagtcgg aatcgcagac
     1921 cgataccagg atcttgccat cctatggaac tgcctcggtg agttttctcc ttcattacag
     1981 aaacggcttt ttcaaaaata tggtattgat aatcctgata tgaataaatt gcagtttcat
     2041 ttgatgctcg atgagttttt ctaatcagaa ttggttaatt ggttgtaaca ttattcagat
     2101 tgggccccgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat
     2161 cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg
     2221 gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga
     2281 gcgcagatac caaatactgt tcttctagtg tagccgtagt taggccacca cttcaagaac
     2341 tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt
     2401 ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag
     2461 cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc
     2521 gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag
     2581 gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca
     2641 gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt
     2701 cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc
     2761 tttttacggt tcctggcctt ttgctggcct tttgctcaca tgtt
//
