LOCUS       sequence-101401-        3074 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  MuLE (Multiple Lentiviral Expression) Entry vector containing a U6 
            promoter for shRNA expression. Compatible with MultiSite Gateway 
            cloning.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3074)
  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 3074)
  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 L1-L4
FEATURES             Location/Qualifiers
     source          1..3074
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    complement(232..327)
                     /gene="mutant version of attL"
                     /label=attL4
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(358..377)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(359..377)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(382..398)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(382..398)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(500..519)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             511..1320
                     /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(578..597)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      1467..2055
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1956..1975
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2209..2226
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(2385..2412)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(2504..2590)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     2639..2661
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     2653..2670
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     2654..2670
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    2686..2785
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        2828..3068
                     /label=U6 promoter
                     /note="Truncated RNA polymerase III promoter for human U6 
                     snRNA used in pSIREN vectors such as 
                     pSIREN-RetroQ-ZsGreen1. Expected transcription start site 
                     is 8bp downstream of this feature."
     primer_bind     2828..2848
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     2999..3018
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
ORIGIN
        1 gatctcggcg ttgtgcatgt cgggcgactg ctccatgatc ttgcgccgct cgatctgcga
       61 ccacaccata aaggcgttca tgggccgctt gatgtggcca ctgggcgtct tgcaccagct
      121 ggggtcgtcc gccttgccgc ccgtcgacgc ggtggagcca ggcgtcgggg aggacgcgat
      181 gcccagctcc aggccggcgc ccgagtccga gctctttttt gaattcgcac ccaacttttc
      241 tatacaaagt tggcattata agaaagcatt gcttatcaat ttgttgcaac gaacaggtca
      301 ctatcagtca aaataaaatc attatttgcc atccagctgc agggcggccg cgatatcccc
      361 tatagtgagt cgtattacat ggtcatagct gtttcctggc agctctggcc cgtgtctcaa
      421 aatctctgat gttacattgc acaagataaa aatatatcat catgaacaat aaaactgtct
      481 gcttacataa acagtaatac aaggggtgtt atgagccata ttcaacggga aacgtcgagg
      541 ccgcgattaa attccaacat ggatgctgat ttatatgggt ataaatgggc tcgcgataat
      601 gtcgggcaat caggtgcgac aatctatcgc ttgtatggga agcccgatgc gccagagttg
      661 tttctgaaac atggcaaagg tagcgttgcc aatgatgtta cagatgagat ggtcagacta
      721 aactggctga cggaatttat gcctcttccg accatcaagc attttatccg tactcctgat
      781 gatgcatggt tactcaccac tgcgatcccc ggaaaaacag cattccaggt attagaagaa
      841 tatcctgatt caggtgaaaa tattgttgat gcgctggcag tgttcctgcg ccggttgcat
      901 tcgattcctg tttgtaattg tccttttaac agcgatcgcg tatttcgtct cgctcaggcg
      961 caatcacgaa tgaataacgg tttggttgat gcgagtgatt ttgatgacga gcgtaatggc
     1021 tggcctgttg aacaagtctg gaaagaaatg cataaacttt tgccattctc accggattca
     1081 gtcgtcactc atggtgattt ctcacttgat aaccttattt ttgacgaggg gaaattaata
     1141 ggttgtattg atgttggacg agtcggaatc gcagaccgat accaggatct tgccatccta
     1201 tggaactgcc tcggtgagtt ttctccttca ttacagaaac ggctttttca aaaatatggt
     1261 attgataatc ctgatatgaa taaattgcag tttcatttga tgctcgatga gtttttctaa
     1321 tcagaattgg ttaattggtt gtaacactgg cagagcatta cgctgacttg acgggacggc
     1381 gcaagctcat gaccaaaatc ccttaacgtg agttacgcgt cgttccactg agcgtcagac
     1441 cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc
     1501 ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca
     1561 actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta
     1621 gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct
     1681 ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg
     1741 gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc
     1801 acacagccca gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta
     1861 tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg
     1921 gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt
     1981 cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg
     2041 cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg
     2101 ccttttgctc acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc
     2161 gcctttgagt gagctgatac cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg
     2221 agcgaggaag cggaagagcg cccaatacgc aaaccgcctc tccccgcgcg ttggccgatt
     2281 cattaatgca gctggcacga caggtttccc gactggaaag cgggcagtga gcgcaacgca
     2341 attaatacgc gtaccgctag ccaggaagag tttgtagaaa cgcaaaaagg ccatccgtca
     2401 ggatggcctt ctgcttagtt tgatgcctgg cagtttatgg cgggcgtcct gcccgccacc
     2461 ctccgggccg ttgcttcaca acgttcaaat ccgctcccgg cggatttgtc ctactcagga
     2521 gagcgttcac cgacaaacaa cagataaaac gaaaggccca gtcttccgac tgagcctttc
     2581 gttttatttg atgcctggca gttccctact ctcgcgttaa cgctagcatg gatgttttcc
     2641 cagtcacgac gttgtaaaac gacggccagt cttaagctcg ggccccaaat aatgatttta
     2701 ttttgactga tagtgacctg ttcgttgcaa caaattgatg agcaatgctt ttttataatg
     2761 ccaactttgt acaaaaaagc aggctctatc gatccgagac tagcctcgag cggccgcccc
     2821 cttcaccgag ggcctatttc ccatgattcc ttcatatttg catatacgat acaaggctgt
     2881 tagagagata attggaatta atttgactgt aaacacaaag atattagtac aaaatacgtg
     2941 acgtagaaag taataatttc ttgggtagtt tgcagtttta aaattatgtt ttaaaatgga
     3001 ctatcatatg cttaccgtaa cttgaaagta tttcgatttc ttggctttat atatcttgtg
     3061 gaaaggacga aaca
//
