LOCUS       sequence-101399-        2850 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 2850)
  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 2850)
  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-L4
FEATURES             Location/Qualifiers
     source          1..2850
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    complement(19..114)
                     /gene="mutant version of attL"
                     /label=attL4
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(145..164)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(146..164)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(169..185)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(169..185)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(287..306)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             298..1107
                     /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(365..384)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      1254..1842
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1743..1762
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1996..2013
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(2172..2199)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(2291..2377)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     2426..2448
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     2440..2457
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     2441..2457
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    2473..2572
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
ORIGIN
        1 cttttttgaa ttcgcaccca acttttctat acaaagttgg cattataaga aagcattgct
       61 tatcaatttg ttgcaacgaa caggtcacta tcagtcaaaa taaaatcatt atttgccatc
      121 cagctgcagg gcggccgcga tatcccctat agtgagtcgt attacatggt catagctgtt
      181 tcctggcagc tctggcccgt gtctcaaaat ctctgatgtt acattgcaca agataaaaat
      241 atatcatcat gaacaataaa actgtctgct tacataaaca gtaatacaag gggtgttatg
      301 agccatattc aacgggaaac gtcgaggccg cgattaaatt ccaacatgga tgctgattta
      361 tatgggtata aatgggctcg cgataatgtc gggcaatcag gtgcgacaat ctatcgcttg
      421 tatgggaagc ccgatgcgcc agagttgttt ctgaaacatg gcaaaggtag cgttgccaat
      481 gatgttacag atgagatggt cagactaaac tggctgacgg aatttatgcc tcttccgacc
      541 atcaagcatt ttatccgtac tcctgatgat gcatggttac tcaccactgc gatccccgga
      601 aaaacagcat tccaggtatt agaagaatat cctgattcag gtgaaaatat tgttgatgcg
      661 ctggcagtgt tcctgcgccg gttgcattcg attcctgttt gtaattgtcc ttttaacagc
      721 gatcgcgtat ttcgtctcgc tcaggcgcaa tcacgaatga ataacggttt ggttgatgcg
      781 agtgattttg atgacgagcg taatggctgg cctgttgaac aagtctggaa agaaatgcat
      841 aaacttttgc cattctcacc ggattcagtc gtcactcatg gtgatttctc acttgataac
      901 cttatttttg acgaggggaa attaataggt tgtattgatg ttggacgagt cggaatcgca
      961 gaccgatacc aggatcttgc catcctatgg aactgcctcg gtgagttttc tccttcatta
     1021 cagaaacggc tttttcaaaa atatggtatt gataatcctg atatgaataa attgcagttt
     1081 catttgatgc tcgatgagtt tttctaatca gaattggtta attggttgta acactggcag
     1141 agcattacgc tgacttgacg ggacggcgca agctcatgac caaaatccct taacgtgagt
     1201 tacgcgtcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat
     1261 cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg
     1321 gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga
     1381 gcgcagatac caaatactgt tcttctagtg tagccgtagt taggccacca cttcaagaac
     1441 tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt
     1501 ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag
     1561 cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc
     1621 gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag
     1681 gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca
     1741 gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt
     1801 cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc
     1861 tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc tgcgttatcc
     1921 cctgattctg tggataaccg tattaccgcc tttgagtgag ctgataccgc tcgccgcagc
     1981 cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg aagagcgccc aatacgcaaa
     2041 ccgcctctcc ccgcgcgttg gccgattcat taatgcagct ggcacgacag gtttcccgac
     2101 tggaaagcgg gcagtgagcg caacgcaatt aatacgcgta ccgctagcca ggaagagttt
     2161 gtagaaacgc aaaaaggcca tccgtcagga tggccttctg cttagtttga tgcctggcag
     2221 tttatggcgg gcgtcctgcc cgccaccctc cgggccgttg cttcacaacg ttcaaatccg
     2281 ctcccggcgg atttgtccta ctcaggagag cgttcaccga caaacaacag ataaaacgaa
     2341 aggcccagtc ttccgactga gcctttcgtt ttatttgatg cctggcagtt ccctactctc
     2401 gcgttaacgc tagcatggat gttttcccag tcacgacgtt gtaaaacgac ggccagtctt
     2461 aagctcgggc cccaaataat gattttattt tgactgatag tgacctgttc gttgcaacaa
     2521 attgatgagc aatgcttttt tataatgcca actttgtaca aaaaagcagg ctctatcgat
     2581 ctagtgctgc agtatttagc atgccccacc catctgcaag gcattctgga tagtgtcaaa
     2641 acagccggaa atcaagtccg tttatctcaa actttagcat tttgggaata aatgatattt
     2701 gctatgctgg ttaaattaga ttttagttaa atttcctgct gaagctctag tacgataagt
     2761 aacttgacct aagtgtaaag ttgagatttc cttcaggttt atatagcttg tgcgccgcct
     2821 gggtacctcg gtcttcacag atctagagct
//
