LOCUS       sequence-112696-        2938 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 2938)
  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 2938)
  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 L5-L2
FEATURES             Location/Qualifiers
     source          1..2938
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    complement(37..136)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(166..185)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(167..185)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(190..206)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(190..206)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(308..327)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             319..1128
                     /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(386..405)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      1275..1863
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1764..1783
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2017..2034
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(2193..2220)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(2312..2398)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     2447..2469
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     2461..2478
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     2462..2478
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    2495..2590
                     /gene="mutant version of attL"
                     /label=attL5
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        2633..2881
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA 
                     (Domitrovich & Kunkel, 2003)"
     primer_bind     2633..2653
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     2804..2823
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
ORIGIN
        1 aattctcgac ctcgagacaa atggcagtga attcgtaccc agctttcttg tacaaagttg
       61 gcattataag aaagcattgc ttatcaattt gttgcaacga acaggtcact atcagtcaaa
      121 ataaaatcat tatttgccat ccagctgcag ggcggccgcg atatccccta tagtgagtcg
      181 tattacatgg tcatagctgt ttcctggcag ctctggcccg tgtctcaaaa tctctgatgt
      241 tacattgcac aagataaaaa tatatcatca tgaacaataa aactgtctgc ttacataaac
      301 agtaatacaa ggggtgttat gagccatatt caacgggaaa cgtcgaggcc gcgattaaat
      361 tccaacatgg atgctgattt atatgggtat aaatgggctc gcgataatgt cgggcaatca
      421 ggtgcgacaa tctatcgctt gtatgggaag cccgatgcgc cagagttgtt tctgaaacat
      481 ggcaaaggta gcgttgccaa tgatgttaca gatgagatgg tcagactaaa ctggctgacg
      541 gaatttatgc ctcttccgac catcaagcat tttatccgta ctcctgatga tgcatggtta
      601 ctcaccactg cgatccccgg aaaaacagca ttccaggtat tagaagaata tcctgattca
      661 ggtgaaaata ttgttgatgc gctggcagtg ttcctgcgcc ggttgcattc gattcctgtt
      721 tgtaattgtc cttttaacag cgatcgcgta tttcgtctcg ctcaggcgca atcacgaatg
      781 aataacggtt tggttgatgc gagtgatttt gatgacgagc gtaatggctg gcctgttgaa
      841 caagtctgga aagaaatgca taaacttttg ccattctcac cggattcagt cgtcactcat
      901 ggtgatttct cacttgataa ccttattttt gacgagggga aattaatagg ttgtattgat
      961 gttggacgag tcggaatcgc agaccgatac caggatcttg ccatcctatg gaactgcctc
     1021 ggtgagtttt ctccttcatt acagaaacgg ctttttcaaa aatatggtat tgataatcct
     1081 gatatgaata aattgcagtt tcatttgatg ctcgatgagt ttttctaatc agaattggtt
     1141 aattggttgt aacactggca gagcattacg ctgacttgac gggacggcgc aagctcatga
     1201 ccaaaatccc ttaacgtgag ttacgcgtcg ttccactgag cgtcagaccc cgtagaaaag
     1261 atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa
     1321 aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg
     1381 aaggtaactg gcttcagcag agcgcagata ccaaatactg ttcttctagt gtagccgtag
     1441 ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg
     1501 ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga
     1561 tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc
     1621 ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc
     1681 acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga
     1741 gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt
     1801 cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg
     1861 aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac
     1921 atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc ctttgagtga
     1981 gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg
     2041 gaagagcgcc caatacgcaa accgcctctc cccgcgcgtt ggccgattca ttaatgcagc
     2101 tggcacgaca ggtttcccga ctggaaagcg ggcagtgagc gcaacgcaat taatacgcgt
     2161 accgctagcc aggaagagtt tgtagaaacg caaaaaggcc atccgtcagg atggccttct
     2221 gcttagtttg atgcctggca gtttatggcg ggcgtcctgc ccgccaccct ccgggccgtt
     2281 gcttcacaac gttcaaatcc gctcccggcg gatttgtcct actcaggaga gcgttcaccg
     2341 acaaacaaca gataaaacga aaggcccagt cttccgactg agcctttcgt tttatttgat
     2401 gcctggcagt tccctactct cgcgttaacg ctagcatgga tgttttccca gtcacgacgt
     2461 tgtaaaacga cggccagtct taagctcggg ccccaaataa tgattttatt ttgactgata
     2521 gtgacctgtt cgttgcaaca aattgatgag caatgctttt ttataatgcc aactttgtat
     2581 acaaaagttg ctatcgatcc gagactagcc tcgagcggcc gcccccttca ccgagggcct
     2641 atttcccatg attccttcat atttgcatat acgatacaag gctgttagag agataattgg
     2701 aattaatttg actgtaaaca caaagatatt agtacaaaat acgtgacgta gaaagtaata
     2761 atttcttggg tagtttgcag ttttaaaatt atgttttaaa atggactatc atatgcttac
     2821 cgtaacttga aagtatttcg atttcttggc tttatatatc ttgtggaaag gacgaaacac
     2881 cggtcctaag gttaagtcgc cctcgctcga gcgagggcga cttaacctta ggtttttg
//
