LOCUS       sequence-101389-        3334 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  MuLE (Multiple Lentiviral Expression) Entry vector containing a 
            tet-inducible CMV promoter and multiple cloning site module. 
            Compatible with MultiSite Gateway cloning.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3334)
  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 3334)
  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 CMV/TO R4-R3
FEATURES             Location/Qualifiers
     source          1..3334
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    93..216
                     /gene="mutant version of attR"
                     /label=attR3
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(271..290)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(272..290)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(295..311)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(295..311)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(413..432)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             424..1233
                     /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(491..510)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      1380..1968
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1869..1888
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2122..2139
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(2298..2325)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(2417..2503)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     2552..2574
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     2566..2583
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     2567..2583
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(2633..2757)
                     /gene="mutant version of attR"
                     /label=attR4
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     enhancer        2774..3077
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        3078..3281
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     3232..3252
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     protein_bind    3283..3301
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    3304..3322
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
ORIGIN
        1 cgagctcgga tccactagta acggccgcca gtgtgctgga attctgcaga tatccatcac
       61 actggcggcc gctcgagcat gcatctagag gacaactttg tataataaag ttgaacgaga
      121 aacgtaaaat gatataaata tcaatatatt aaattagatt ttgcataaaa aacagactac
      181 ataatactgt aaaacacaac atatccagtc actatgaatc aactacttag atggtattag
      241 tgacctgtac tgcagggcgg ccgcgatatc ccctatagtg agtcgtatta catggtcata
      301 gctgtttcct ggcagctctg gcccgtgtct caaaatctct gatgttacat tgcacaagat
      361 aaaaatatat catcatgaac aataaaactg tctgcttaca taaacagtaa tacaaggggt
      421 gttatgagcc atattcaacg ggaaacgtcg aggccgcgat taaattccaa catggatgct
      481 gatttatatg ggtataaatg ggctcgcgat aatgtcgggc aatcaggtgc gacaatctat
      541 cgcttgtatg ggaagcccga tgcgccagag ttgtttctga aacatggcaa aggtagcgtt
      601 gccaatgatg ttacagatga gatggtcaga ctaaactggc tgacggaatt tatgcctctt
      661 ccgaccatca agcattttat ccgtactcct gatgatgcat ggttactcac cactgcgatc
      721 cccggaaaaa cagcattcca ggtattagaa gaatatcctg attcaggtga aaatattgtt
      781 gatgcgctgg cagtgttcct gcgccggttg cattcgattc ctgtttgtaa ttgtcctttt
      841 aacagcgatc gcgtatttcg tctcgctcag gcgcaatcac gaatgaataa cggtttggtt
      901 gatgcgagtg attttgatga cgagcgtaat ggctggcctg ttgaacaagt ctggaaagaa
      961 atgcataaac ttttgccatt ctcaccggat tcagtcgtca ctcatggtga tttctcactt
     1021 gataacctta tttttgacga ggggaaatta ataggttgta ttgatgttgg acgagtcgga
     1081 atcgcagacc gataccagga tcttgccatc ctatggaact gcctcggtga gttttctcct
     1141 tcattacaga aacggctttt tcaaaaatat ggtattgata atcctgatat gaataaattg
     1201 cagtttcatt tgatgctcga tgagtttttc taatcagaat tggttaattg gttgtaacac
     1261 tggcagagca ttacgctgac ttgacgggac ggcgcaagct catgaccaaa atcccttaac
     1321 gtgagttacg cgtcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct
     1381 tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca
     1441 gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc
     1501 agcagagcgc agataccaaa tactgttctt ctagtgtagc cgtagttagg ccaccacttc
     1561 aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct
     1621 gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag
     1681 gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc
     1741 tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg
     1801 agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag
     1861 cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt
     1921 gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac
     1981 gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt ctttcctgcg
     2041 ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga taccgctcgc
     2101 cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga gcgcccaata
     2161 cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctggca cgacaggttt
     2221 cccgactgga aagcgggcag tgagcgcaac gcaattaata cgcgtaccgc tagccaggaa
     2281 gagtttgtag aaacgcaaaa aggccatccg tcaggatggc cttctgctta gtttgatgcc
     2341 tggcagttta tggcgggcgt cctgcccgcc accctccggg ccgttgcttc acaacgttca
     2401 aatccgctcc cggcggattt gtcctactca ggagagcgtt caccgacaaa caacagataa
     2461 aacgaaaggc ccagtcttcc gactgagcct ttcgttttat ttgatgcctg gcagttccct
     2521 actctcgcgt taacgctagc atggatgttt tcccagtcac gacgttgtaa aacgacggcc
     2581 agtcttaagc tcgggcccct acaggtcact aataccatct aagtagttga ttcatagtga
     2641 ctggatatgt tgtgttttac agtattatgt agtctgtttt ttatgcaaaa tctaatttaa
     2701 tatattgata tttatatcat tttacgtttc tcgttcaact tttctataca aagttgggag
     2761 acccaagctt ccgcgttaca taacttacgg taaatggccc gcctggctga ccgcccaacg
     2821 acccccgccc attgacgtca ataatgacgt atgttcccat agtaacgcca atagggactt
     2881 tccattgacg tcaatgggtg gagtatttac ggtaaactgc ccacttggca gtacatcaag
     2941 tgtatcatat gccaagtacg ccccctattg acgtcaatga cggtaaatgg cccgcctggc
     3001 attatgccca gtacatgacc ttatgggact ttcctacttg gcagtacatc tacgtattag
     3061 tcatcgctat taccatggtg atgcggtttt ggcagtacat caatgggcgt ggatagcggt
     3121 ttgactcacg gggatttcca agtctccacc ccattgacgt caatgggagt ttgttttggc
     3181 accaaaatca acgggacttt ccaaaatgtc gtaacaactc cgccccattg acgcaaatgg
     3241 gcggtaggcg tgtacggtgg gaggtctata taagcagagc tctccctatc agtgatagag
     3301 atctccctat cagtgataga gatcgtcgag gtac
//
