LOCUS       sequence-101395-        2899 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  MuLE (Multiple Lentiviral Expression) Entry vector containing a 7SK 
            RNA polymerase III 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 2899)
  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 2899)
  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 7SK L1-R5
FEATURES             Location/Qualifiers
     source          1..2899
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    16..139
                     /gene="mutant version of attR"
                     /label=attR5
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(194..213)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(195..213)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(218..234)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(218..234)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(336..355)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             347..1156
                     /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(414..433)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      1303..1891
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1792..1811
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2045..2062
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(2221..2248)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(2340..2426)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     2475..2497
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     2489..2506
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     2490..2506
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    2522..2621
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
ORIGIN
        1 cttttttgaa ttcgacaact ttgtatacaa aagttgaacg agaaacgtaa aatgatataa
       61 atatcaatat attaaattag attttgcata aaaaacagac tacataatac tgtaaaacac
      121 aacatatcca gtcactatga atcaactact tagatggtat tagtgacctg tactgcaggg
      181 cggccgcgat atcccctata gtgagtcgta ttacatggtc atagctgttt cctggcagct
      241 ctggcccgtg tctcaaaatc tctgatgtta cattgcacaa gataaaaata tatcatcatg
      301 aacaataaaa ctgtctgctt acataaacag taatacaagg ggtgttatga gccatattca
      361 acgggaaacg tcgaggccgc gattaaattc caacatggat gctgatttat atgggtataa
      421 atgggctcgc gataatgtcg ggcaatcagg tgcgacaatc tatcgcttgt atgggaagcc
      481 cgatgcgcca gagttgtttc tgaaacatgg caaaggtagc gttgccaatg atgttacaga
      541 tgagatggtc agactaaact ggctgacgga atttatgcct cttccgacca tcaagcattt
      601 tatccgtact cctgatgatg catggttact caccactgcg atccccggaa aaacagcatt
      661 ccaggtatta gaagaatatc ctgattcagg tgaaaatatt gttgatgcgc tggcagtgtt
      721 cctgcgccgg ttgcattcga ttcctgtttg taattgtcct tttaacagcg atcgcgtatt
      781 tcgtctcgct caggcgcaat cacgaatgaa taacggtttg gttgatgcga gtgattttga
      841 tgacgagcgt aatggctggc ctgttgaaca agtctggaaa gaaatgcata aacttttgcc
      901 attctcaccg gattcagtcg tcactcatgg tgatttctca cttgataacc ttatttttga
      961 cgaggggaaa ttaataggtt gtattgatgt tggacgagtc ggaatcgcag accgatacca
     1021 ggatcttgcc atcctatgga actgcctcgg tgagttttct ccttcattac agaaacggct
     1081 ttttcaaaaa tatggtattg ataatcctga tatgaataaa ttgcagtttc atttgatgct
     1141 cgatgagttt ttctaatcag aattggttaa ttggttgtaa cactggcaga gcattacgct
     1201 gacttgacgg gacggcgcaa gctcatgacc aaaatccctt aacgtgagtt acgcgtcgtt
     1261 ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct
     1321 gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc
     1381 ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc
     1441 aaatactgtt cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc
     1501 gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc
     1561 gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg
     1621 aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata
     1681 cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta
     1741 tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc
     1801 ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg
     1861 atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt
     1921 cctggccttt tgctggcctt ttgctcacat gttctttcct gcgttatccc ctgattctgt
     1981 ggataaccgt attaccgcct ttgagtgagc tgataccgct cgccgcagcc gaacgaccga
     2041 gcgcagcgag tcagtgagcg aggaagcgga agagcgccca atacgcaaac cgcctctccc
     2101 cgcgcgttgg ccgattcatt aatgcagctg gcacgacagg tttcccgact ggaaagcggg
     2161 cagtgagcgc aacgcaatta atacgcgtac cgctagccag gaagagtttg tagaaacgca
     2221 aaaaggccat ccgtcaggat ggccttctgc ttagtttgat gcctggcagt ttatggcggg
     2281 cgtcctgccc gccaccctcc gggccgttgc ttcacaacgt tcaaatccgc tcccggcgga
     2341 tttgtcctac tcaggagagc gttcaccgac aaacaacaga taaaacgaaa ggcccagtct
     2401 tccgactgag cctttcgttt tatttgatgc ctggcagttc cctactctcg cgttaacgct
     2461 agcatggatg ttttcccagt cacgacgttg taaaacgacg gccagtctta agctcgggcc
     2521 ccaaataatg attttatttt gactgatagt gacctgttcg ttgcaacaaa ttgatgagca
     2581 atgctttttt ataatgccaa ctttgtacaa aaaagcaggc tctatcgatc tagtgctgca
     2641 gtatttagca tgccccaccc atctgcaagg cattctggat agtgtcaaaa cagccggaaa
     2701 tcaagtccgt ttatctcaaa ctttagcatt ttgggaataa atgatatttg ctatgctggt
     2761 taaattagat tttagttaaa tttcctgctg aagctctagt acgataagta acttgaccta
     2821 agtgtaaagt tgagatttcc ttcaggttta tatagcttgt gcgccgcctg ggtacctcgg
     2881 tcttcacaga tctagagct
//
