LOCUS       sequence-101393-        3048 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  MuLE (Multiple Lentiviral Expression) Entry vector containing a H1 
            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 3048)
  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 3048)
  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 H1 L1-L4
FEATURES             Location/Qualifiers
     source          1..3048
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     92..109
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(268..295)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(387..473)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     522..544
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     536..553
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     537..553
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    569..668
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        716..930
                     /label=H1 promoter
                     /note="human H1 RNA promoter"
     primer_bind     851..870
                     /label=H1
                     /note="Human H1 promoter, forward primer"
     protein_bind    complement(1163..1258)
                     /gene="mutant version of attL"
                     /label=attL4
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1289..1308)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1290..1308)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1313..1329)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1313..1329)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1431..1450)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             1442..2251
                     /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(1509..1528)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      2398..2986
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2887..2906
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga
       61 taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga
      121 gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctggca
      181 cgacaggttt cccgactgga aagcgggcag tgagcgcaac gcaattaata cgcgtaccgc
      241 tagccaggaa gagtttgtag aaacgcaaaa aggccatccg tcaggatggc cttctgctta
      301 gtttgatgcc tggcagttta tggcgggcgt cctgcccgcc accctccggg ccgttgcttc
      361 acaacgttca aatccgctcc cggcggattt gtcctactca ggagagcgtt caccgacaaa
      421 caacagataa aacgaaaggc ccagtcttcc gactgagcct ttcgttttat ttgatgcctg
      481 gcagttccct actctcgcgt taacgctagc atggatgttt tcccagtcac gacgttgtaa
      541 aacgacggcc agtcttaagc tcgggcccca aataatgatt ttattttgac tgatagtgac
      601 ctgttcgttg caacaaattg atgagcaatg cttttttata atgccaactt tgtacaaaaa
      661 agcaggctct atcgatccaa ttcagtcgac tggatccccc gggctgcagg aattcgaacg
      721 ctgacgtcat caacccgctc caaggaatcg cgggcccagt gtcactaggc gggaacaccc
      781 agcgcgcgtg cgccctggca ggaagatggc tgtgagggac aggggagtgg cgccctgcaa
      841 tatttgcatg tcgctatgtg ttctgggaaa tcaccataaa cgtgaaatgt ctttggattt
      901 gggaatctta taagttctgt atgagaccac agatctcggc gttgtgcatg tcgggcgact
      961 gctccatgat cttgcgccgc tcgatctgcg accacaccat aaaggcgttc atgggccgct
     1021 tgatgtggcc actgggcgtc ttgcaccagc tggggtcgtc cgccttgccg cccgtcgacg
     1081 cggtggagcc aggcgtcggg gaggacgcga tgcccagctc caggccggcg cccgagtccg
     1141 agctcttttt tgaattcgca cccaactttt ctatacaaag ttggcattat aagaaagcat
     1201 tgcttatcaa tttgttgcaa cgaacaggtc actatcagtc aaaataaaat cattatttgc
     1261 catccagctg cagggcggcc gcgatatccc ctatagtgag tcgtattaca tggtcatagc
     1321 tgtttcctgg cagctctggc ccgtgtctca aaatctctga tgttacattg cacaagataa
     1381 aaatatatca tcatgaacaa taaaactgtc tgcttacata aacagtaata caaggggtgt
     1441 tatgagccat attcaacggg aaacgtcgag gccgcgatta aattccaaca tggatgctga
     1501 tttatatggg tataaatggg ctcgcgataa tgtcgggcaa tcaggtgcga caatctatcg
     1561 cttgtatggg aagcccgatg cgccagagtt gtttctgaaa catggcaaag gtagcgttgc
     1621 caatgatgtt acagatgaga tggtcagact aaactggctg acggaattta tgcctcttcc
     1681 gaccatcaag cattttatcc gtactcctga tgatgcatgg ttactcacca ctgcgatccc
     1741 cggaaaaaca gcattccagg tattagaaga atatcctgat tcaggtgaaa atattgttga
     1801 tgcgctggca gtgttcctgc gccggttgca ttcgattcct gtttgtaatt gtccttttaa
     1861 cagcgatcgc gtatttcgtc tcgctcaggc gcaatcacga atgaataacg gtttggttga
     1921 tgcgagtgat tttgatgacg agcgtaatgg ctggcctgtt gaacaagtct ggaaagaaat
     1981 gcataaactt ttgccattct caccggattc agtcgtcact catggtgatt tctcacttga
     2041 taaccttatt tttgacgagg ggaaattaat aggttgtatt gatgttggac gagtcggaat
     2101 cgcagaccga taccaggatc ttgccatcct atggaactgc ctcggtgagt tttctccttc
     2161 attacagaaa cggctttttc aaaaatatgg tattgataat cctgatatga ataaattgca
     2221 gtttcatttg atgctcgatg agtttttcta atcagaattg gttaattggt tgtaacactg
     2281 gcagagcatt acgctgactt gacgggacgg cgcaagctca tgaccaaaat cccttaacgt
     2341 gagttacgcg tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg
     2401 agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc
     2461 ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag
     2521 cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttaggcc accacttcaa
     2581 gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc
     2641 cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc
     2701 gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta
     2761 caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag
     2821 aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct
     2881 tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga
     2941 gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc
     3001 ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgtt
//
