LOCUS       sequence-101383-        3411 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  MuLE (Multiple Lentiviral Expression) Entry vector containing a CMV 
            promoter and multiple cloning site flanked by loxP sites. Compatible
            with MultiSite Gateway cloning.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3411)
  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 3411)
  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 loxP L5-L2
FEATURES             Location/Qualifiers
     source          1..3411
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    complement(7..40)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     protein_bind    complement(132..165)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     protein_bind    complement(176..275)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        complement(306..324)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(329..345)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(329..345)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(447..466)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             458..1267
                     /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(525..544)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      1414..2002
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1903..1922
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2156..2173
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(2332..2359)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(2451..2537)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     2586..2608
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     2600..2617
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     2601..2617
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    2634..2729
                     /gene="mutant version of attL"
                     /label=attL5
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     enhancer        2757..3136
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        3137..3340
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     3291..3311
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     promoter        3385..3403
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
        1 agcttcataa cttcgtatag catacattat acgaagttat gcttggtacc gagctcggat
       61 ccactagtaa cggccgccag tgtgctggaa ttctgcagat atccatcaca ctggcggccg
      121 ctcgagcatg cataacttcg tatagcatac attatacgaa gttatgtcta gaggtaccca
      181 gctttcttgt acaaagttgg cattataaga aagcattgct tatcaatttg ttgcaacgaa
      241 caggtcacta tcagtcaaaa taaaatcatt atttgccatc cagctgcagg gcggccgcga
      301 tatcccctat agtgagtcgt attacatggt catagctgtt tcctggcagc tctggcccgt
      361 gtctcaaaat ctctgatgtt acattgcaca agataaaaat atatcatcat gaacaataaa
      421 actgtctgct tacataaaca gtaatacaag gggtgttatg agccatattc aacgggaaac
      481 gtcgaggccg cgattaaatt ccaacatgga tgctgattta tatgggtata aatgggctcg
      541 cgataatgtc gggcaatcag gtgcgacaat ctatcgcttg tatgggaagc ccgatgcgcc
      601 agagttgttt ctgaaacatg gcaaaggtag cgttgccaat gatgttacag atgagatggt
      661 cagactaaac tggctgacgg aatttatgcc tcttccgacc atcaagcatt ttatccgtac
      721 tcctgatgat gcatggttac tcaccactgc gatccccgga aaaacagcat tccaggtatt
      781 agaagaatat cctgattcag gtgaaaatat tgttgatgcg ctggcagtgt tcctgcgccg
      841 gttgcattcg attcctgttt gtaattgtcc ttttaacagc gatcgcgtat ttcgtctcgc
      901 tcaggcgcaa tcacgaatga ataacggttt ggttgatgcg agtgattttg atgacgagcg
      961 taatggctgg cctgttgaac aagtctggaa agaaatgcat aaacttttgc cattctcacc
     1021 ggattcagtc gtcactcatg gtgatttctc acttgataac cttatttttg acgaggggaa
     1081 attaataggt tgtattgatg ttggacgagt cggaatcgca gaccgatacc aggatcttgc
     1141 catcctatgg aactgcctcg gtgagttttc tccttcatta cagaaacggc tttttcaaaa
     1201 atatggtatt gataatcctg atatgaataa attgcagttt catttgatgc tcgatgagtt
     1261 tttctaatca gaattggtta attggttgta acactggcag agcattacgc tgacttgacg
     1321 ggacggcgca agctcatgac caaaatccct taacgtgagt tacgcgtcgt tccactgagc
     1381 gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat
     1441 ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga
     1501 gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt
     1561 tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata
     1621 cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac
     1681 cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg
     1741 ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg
     1801 tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag
     1861 cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct
     1921 ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc
     1981 aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt
     2041 ttgctggcct tttgctcaca tgttctttcc tgcgttatcc cctgattctg tggataaccg
     2101 tattaccgcc tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga
     2161 gtcagtgagc gaggaagcgg aagagcgccc aatacgcaaa ccgcctctcc ccgcgcgttg
     2221 gccgattcat taatgcagct ggcacgacag gtttcccgac tggaaagcgg gcagtgagcg
     2281 caacgcaatt aatacgcgta ccgctagcca ggaagagttt gtagaaacgc aaaaaggcca
     2341 tccgtcagga tggccttctg cttagtttga tgcctggcag tttatggcgg gcgtcctgcc
     2401 cgccaccctc cgggccgttg cttcacaacg ttcaaatccg ctcccggcgg atttgtccta
     2461 ctcaggagag cgttcaccga caaacaacag ataaaacgaa aggcccagtc ttccgactga
     2521 gcctttcgtt ttatttgatg cctggcagtt ccctactctc gcgttaacgc tagcatggat
     2581 gttttcccag tcacgacgtt gtaaaacgac ggccagtctt aagctcgggc cccaaataat
     2641 gattttattt tgactgatag tgacctgttc gttgcaacaa attgatgagc aatgcttttt
     2701 tataatgcca actttgtata caaaagttgg cgatgtacgg gccagatata cgcgttgaca
     2761 ttgattattg actagttatt aatagtaatc aattacgggg tcattagttc atagcccata
     2821 tatggagttc cgcgttacat aacttacggt aaatggcccg cctggctgac cgcccaacga
     2881 cccccgccca ttgacgtcaa taatgacgta tgttcccata gtaacgccaa tagggacttt
     2941 ccattgacgt caatgggtgg agtatttacg gtaaactgcc cacttggcag tacatcaagt
     3001 gtatcatatg ccaagtacgc cccctattga cgtcaatgac ggtaaatggc ccgcctggca
     3061 ttatgcccag tacatgacct tatgggactt tcctacttgg cagtacatct acgtattagt
     3121 catcgctatt accatggtga tgcggttttg gcagtacatc aatgggcgtg gatagcggtt
     3181 tgactcacgg ggatttccaa gtctccaccc cattgacgtc aatgggagtt tgttttggca
     3241 ccaaaatcaa cgggactttc caaaatgtcg taacaactcc gccccattga cgcaaatggg
     3301 cggtaggcgt gtacggtggg aggtctatat aagcagagct ctctggctaa ctagagaacc
     3361 cactgcttac tggcttatcg aaattaatac gactcactat agggagaccc a
//
