LOCUS       sequence-101420-        2912 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  MuLE (Multiple Lentiviral Expression) Entry vector containing a U6 
            promoter and a non-silencing (scrambled) shRNA module. Compatible 
            with MultiSite Gateway cloning.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2912)
  AUTHORS   Albers J, Danzer C, Rechsteiner M, Lehmann H, Brandt LP, Hejhal T, 
            Catalano A, Busenhart P, Goncalves AF, Brandt S, Bode PK, 
            Bode-Lesniewska B, Wild PJ, Frew IJ
  TITLE     A versatile modular vector system for rapid combinatorial mammalian 
            genetics.
  JOURNAL   J Clin Invest. 2015 Mar 9. pii: 79743. doi: 10.1172/JCI79743.
  PUBMED    25751063
REFERENCE   2  (bases 1 to 2912)
  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: "J Clin 
            Invest. 2015 Mar 9. pii: 79743. doi: 10.1172/JCI79743."
COMMENT     Sequence Label: pMuLE ENTR U6 n.s shRNA L1-L4
FEATURES             Location/Qualifiers
     source          1..2912
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    complement(11..106)
                     /gene="mutant version of attL"
                     /label=attL4
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(137..156)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(138..156)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(161..177)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(161..177)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(279..298)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             290..1099
                     /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
                     FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
                     FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
                     RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     complement(357..376)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      1246..1834
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1735..1754
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1988..2005
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(2164..2191)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(2283..2369)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     2418..2440
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     2432..2449
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     2433..2449
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    2465..2564
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        2607..2856
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     2607..2627
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     2779..2798
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
ORIGIN
        1 aattcgcacc caacttttct atacaaagtt ggcattataa gaaagcattg cttatcaatt
       61 tgttgcaacg aacaggtcac tatcagtcaa aataaaatca ttatttgcca tccagctgca
      121 gggcggccgc gatatcccct atagtgagtc gtattacatg gtcatagctg tttcctggca
      181 gctctggccc gtgtctcaaa atctctgatg ttacattgca caagataaaa atatatcatc
      241 atgaacaata aaactgtctg cttacataaa cagtaataca aggggtgtta tgagccatat
      301 tcaacgggaa acgtcgaggc cgcgattaaa ttccaacatg gatgctgatt tatatgggta
      361 taaatgggct cgcgataatg tcgggcaatc aggtgcgaca atctatcgct tgtatgggaa
      421 gcccgatgcg ccagagttgt ttctgaaaca tggcaaaggt agcgttgcca atgatgttac
      481 agatgagatg gtcagactaa actggctgac ggaatttatg cctcttccga ccatcaagca
      541 ttttatccgt actcctgatg atgcatggtt actcaccact gcgatccccg gaaaaacagc
      601 attccaggta ttagaagaat atcctgattc aggtgaaaat attgttgatg cgctggcagt
      661 gttcctgcgc cggttgcatt cgattcctgt ttgtaattgt ccttttaaca gcgatcgcgt
      721 atttcgtctc gctcaggcgc aatcacgaat gaataacggt ttggttgatg cgagtgattt
      781 tgatgacgag cgtaatggct ggcctgttga acaagtctgg aaagaaatgc ataaactttt
      841 gccattctca ccggattcag tcgtcactca tggtgatttc tcacttgata accttatttt
      901 tgacgagggg aaattaatag gttgtattga tgttggacga gtcggaatcg cagaccgata
      961 ccaggatctt gccatcctat ggaactgcct cggtgagttt tctccttcat tacagaaacg
     1021 gctttttcaa aaatatggta ttgataatcc tgatatgaat aaattgcagt ttcatttgat
     1081 gctcgatgag tttttctaat cagaattggt taattggttg taacactggc agagcattac
     1141 gctgacttga cgggacggcg caagctcatg accaaaatcc cttaacgtga gttacgcgtc
     1201 gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt
     1261 tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt
     1321 gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat
     1381 accaaatact gttcttctag tgtagccgta gttaggccac cacttcaaga actctgtagc
     1441 accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa
     1501 gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg
     1561 ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag
     1621 atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag
     1681 gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa
     1741 cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt
     1801 gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg
     1861 gttcctggcc ttttgctggc cttttgctca catgttcttt cctgcgttat cccctgattc
     1921 tgtggataac cgtattaccg cctttgagtg agctgatacc gctcgccgca gccgaacgac
     1981 cgagcgcagc gagtcagtga gcgaggaagc ggaagagcgc ccaatacgca aaccgcctct
     2041 ccccgcgcgt tggccgattc attaatgcag ctggcacgac aggtttcccg actggaaagc
     2101 gggcagtgag cgcaacgcaa ttaatacgcg taccgctagc caggaagagt ttgtagaaac
     2161 gcaaaaaggc catccgtcag gatggccttc tgcttagttt gatgcctggc agtttatggc
     2221 gggcgtcctg cccgccaccc tccgggccgt tgcttcacaa cgttcaaatc cgctcccggc
     2281 ggatttgtcc tactcaggag agcgttcacc gacaaacaac agataaaacg aaaggcccag
     2341 tcttccgact gagcctttcg ttttatttga tgcctggcag ttccctactc tcgcgttaac
     2401 gctagcatgg atgttttccc agtcacgacg ttgtaaaacg acggccagtc ttaagctcgg
     2461 gccccaaata atgattttat tttgactgat agtgacctgt tcgttgcaac aaattgatga
     2521 gcaatgcttt tttataatgc caactttgta caaaaaagca ggctctatcg atccgagact
     2581 agcctcgagc ggccgccccc ttcaccgagg gcctatttcc catgattcct tcatatttgc
     2641 atatacgnnt acaaggctgt tagagagata attggaatta atttgactgt aaacacaaag
     2701 atattagtac aaaatacgtg acgtagaaag taataatttc ttgggtagtt tgcagtttta
     2761 aaattatgtt ttaaaatgga ctatcatatg cttaccgtaa cttgaaagta tttcgatttc
     2821 ttggctttat atatcttgtg gaaaggacga aacaccggtc taaggttaag tcgccctcgc
     2881 tcgagcgagg gcgacttaac cttaggtttn tg
//
