LOCUS       sequence-101405-        3209 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  MuLE (Multiple Lentiviral Expression) Entry vector containing a 7SK 
            RNA polymerase III promoter and miR30-based hairpin module for shRNA
            expression. Compatible with MultiSite Gateway cloning.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3209)
  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 3209)
  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-miR-30 R4-R3
FEATURES             Location/Qualifiers
     source          1..3209
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    8..131
                     /gene="mutant version of attR"
                     /label=attR3
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(186..205)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(187..205)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(210..226)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(210..226)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(328..347)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             339..1148
                     /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(406..425)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      1295..1883
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1784..1803
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2037..2054
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(2213..2240)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(2332..2418)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     2467..2489
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     2481..2498
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     2482..2498
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(2548..2671)
                     /gene="mutant version of attR"
                     /label=attR4
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     ncRNA           2986..3079
                     /label=5' miR-30a
                     /note="sequence upstream of the 71-nt precursor of the 
                     human miR-30a microRNA (Zeng et al., 2002)"
                     /ncRNA_class="miRNA"
     ncRNA           3093..3205
                     /label=3' miR-30a
                     /note="sequence downstream of the 71-nt precursor of the 
                     human miR-30a microRNA (Zeng et al., 2002)"
                     /ncRNA_class="miRNA"
ORIGIN
        1 aattcgacaa ctttgtataa taaagttgaa cgagaaacgt aaaatgatat aaatatcaat
       61 atattaaatt agattttgca taaaaaacag actacataat actgtaaaac acaacatatc
      121 cagtcactat gaatcaacta cttagatggt attagtgacc tgtactgcag ggcggccgcg
      181 atatccccta tagtgagtcg tattacatgg tcatagctgt ttcctggcag ctctggcccg
      241 tgtctcaaaa tctctgatgt tacattgcac aagataaaaa tatatcatca tgaacaataa
      301 aactgtctgc ttacataaac agtaatacaa ggggtgttat gagccatatt caacgggaaa
      361 cgtcgaggcc gcgattaaat tccaacatgg atgctgattt atatgggtat aaatgggctc
      421 gcgataatgt cgggcaatca ggtgcgacaa tctatcgctt gtatgggaag cccgatgcgc
      481 cagagttgtt tctgaaacat ggcaaaggta gcgttgccaa tgatgttaca gatgagatgg
      541 tcagactaaa ctggctgacg gaatttatgc ctcttccgac catcaagcat tttatccgta
      601 ctcctgatga tgcatggtta ctcaccactg cgatccccgg aaaaacagca ttccaggtat
      661 tagaagaata tcctgattca ggtgaaaata ttgttgatgc gctggcagtg ttcctgcgcc
      721 ggttgcattc gattcctgtt tgtaattgtc cttttaacag cgatcgcgta tttcgtctcg
      781 ctcaggcgca atcacgaatg aataacggtt tggttgatgc gagtgatttt gatgacgagc
      841 gtaatggctg gcctgttgaa caagtctgga aagaaatgca taaacttttg ccattctcac
      901 cggattcagt cgtcactcat ggtgatttct cacttgataa ccttattttt gacgagggga
      961 aattaatagg ttgtattgat gttggacgag tcggaatcgc agaccgatac caggatcttg
     1021 ccatcctatg gaactgcctc ggtgagtttt ctccttcatt acagaaacgg ctttttcaaa
     1081 aatatggtat tgataatcct gatatgaata aattgcagtt tcatttgatg ctcgatgagt
     1141 ttttctaatc agaattggtt aattggttgt aacactggca gagcattacg ctgacttgac
     1201 gggacggcgc aagctcatga ccaaaatccc ttaacgtgag ttacgcgtcg ttccactgag
     1261 cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa
     1321 tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag
     1381 agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg
     1441 ttcttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat
     1501 acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta
     1561 ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg
     1621 gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc
     1681 gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa
     1741 gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc
     1801 tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt
     1861 caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct
     1921 tttgctggcc ttttgctcac atgttctttc ctgcgttatc ccctgattct gtggataacc
     1981 gtattaccgc ctttgagtga gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg
     2041 agtcagtgag cgaggaagcg gaagagcgcc caatacgcaa accgcctctc cccgcgcgtt
     2101 ggccgattca ttaatgcagc tggcacgaca ggtttcccga ctggaaagcg ggcagtgagc
     2161 gcaacgcaat taatacgcgt accgctagcc aggaagagtt tgtagaaacg caaaaaggcc
     2221 atccgtcagg atggccttct gcttagtttg atgcctggca gtttatggcg ggcgtcctgc
     2281 ccgccaccct ccgggccgtt gcttcacaac gttcaaatcc gctcccggcg gatttgtcct
     2341 actcaggaga gcgttcaccg acaaacaaca gataaaacga aaggcccagt cttccgactg
     2401 agcctttcgt tttatttgat gcctggcagt tccctactct cgcgttaacg ctagcatgga
     2461 tgttttccca gtcacgacgt tgtaaaacga cggccagtct taagctcggg cccctacagg
     2521 tcactaatac catctaagta gttgattcat agtgactgga tatgttgtgt tttacagtat
     2581 tatgtagtct gttttttatg caaaatctaa tttaatatat tgatatttat atcattttac
     2641 gtttctcgtt caacttttct atacaaagtt gctatcgatc tagtgctgca gtatttagca
     2701 tgccccaccc atctgcaagg cattctggat agtgtcaaaa cagccggaaa tcaagtccgt
     2761 ttatctcaaa ctttagcatt ttgggaataa atgatatttg ctatgctggt taaattagat
     2821 tttagttaaa tttcctgctg aagctctagt acgataagta acttgaccta agtgtaaagt
     2881 tgagatttcc ttcaggttta tatagcttgt gcgccgcctg ggtacctcgg tcttcataga
     2941 tctgcagcac atatactagt cgactaggga taacagggta attgtttgaa tgaggcttca
     3001 gtactttaca gaatcgttgc ctgcacatct tggaaacact tgctgggatt acttcttcag
     3061 gttaacccaa cagaaggctc gagcaaccag aattcagggg gctactttag gagcaattat
     3121 cttgtttact aaaactgaat accttgctat ctctttgata catttttaca aagctgaatt
     3181 aaaatggtat aaattaaatc acttttttc
//
