LOCUS       sequence-347564-        2665 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  MuLE (Multiple Lentiviral Expression) Entry vector containing a 
            multiple cloning site. Compatible with MultiSite Gateway cloning.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2665)
  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 2665)
  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 MCS L5-L4
FEATURES             Location/Qualifiers
     source          1..2665
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      complement(169..196)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(288..374)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     423..445
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     437..454
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     438..454
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    471..566
                     /gene="mutant version of attL"
                     /label=attL5
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     protein_bind    complement(681..776)
                     /gene="mutant version of attL"
                     /label=attL4
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(807..826)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(808..826)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(831..847)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(831..847)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(949..968)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             960..1769
                     /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(1027..1046)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      1916..2504
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2405..2424
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     join(2658..2665,1..10)
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 agtgagcgag gaagcggaag agcgcccaat acgcaaaccg cctctccccg cgcgttggcc
       61 gattcattaa tgcagctggc acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa
      121 cgcaattaat acgcgtaccg ctagccagga agagtttgta gaaacgcaaa aaggccatcc
      181 gtcaggatgg ccttctgctt agtttgatgc ctggcagttt atggcgggcg tcctgcccgc
      241 caccctccgg gccgttgctt cacaacgttc aaatccgctc ccggcggatt tgtcctactc
      301 aggagagcgt tcaccgacaa acaacagata aaacgaaagg cccagtcttc cgactgagcc
      361 tttcgtttta tttgatgcct ggcagttccc tactctcgcg ttaacgctag catggatgtt
      421 ttcccagtca cgacgttgta aaacgacggc cagtcttaag ctcgggcccc aaataatgat
      481 tttattttga ctgatagtga cctgttcgtt gcaacaaatt gatgagcaat gcttttttat
      541 aatgccaact ttgtatacaa aagttgggag acccaagctt ggtaccgagc tcggatccac
      601 tagtaacggc cgccagtgtg ctggaattct gcagatatcc atcacactgg cggccgctcg
      661 agcatgcatc tagaggcacc caacttttct atacaaagtt ggcattataa gaaagcattg
      721 cttatcaatt tgttgcaacg aacaggtcac tatcagtcaa aataaaatca ttatttgcca
      781 tccagctgca gggcggccgc gatatcccct atagtgagtc gtattacatg gtcatagctg
      841 tttcctggca gctctggccc gtgtctcaaa atctctgatg ttacattgca caagataaaa
      901 atatatcatc atgaacaata aaactgtctg cttacataaa cagtaataca aggggtgtta
      961 tgagccatat tcaacgggaa acgtcgaggc cgcgattaaa ttccaacatg gatgctgatt
     1021 tatatgggta taaatgggct cgcgataatg tcgggcaatc aggtgcgaca atctatcgct
     1081 tgtatgggaa gcccgatgcg ccagagttgt ttctgaaaca tggcaaaggt agcgttgcca
     1141 atgatgttac agatgagatg gtcagactaa actggctgac ggaatttatg cctcttccga
     1201 ccatcaagca ttttatccgt actcctgatg atgcatggtt actcaccact gcgatccccg
     1261 gaaaaacagc attccaggta ttagaagaat atcctgattc aggtgaaaat attgttgatg
     1321 cgctggcagt gttcctgcgc cggttgcatt cgattcctgt ttgtaattgt ccttttaaca
     1381 gcgatcgcgt atttcgtctc gctcaggcgc aatcacgaat gaataacggt ttggttgatg
     1441 cgagtgattt tgatgacgag cgtaatggct ggcctgttga acaagtctgg aaagaaatgc
     1501 ataaactttt gccattctca ccggattcag tcgtcactca tggtgatttc tcacttgata
     1561 accttatttt tgacgagggg aaattaatag gttgtattga tgttggacga gtcggaatcg
     1621 cagaccgata ccaggatctt gccatcctat ggaactgcct cggtgagttt tctccttcat
     1681 tacagaaacg gctttttcaa aaatatggta ttgataatcc tgatatgaat aaattgcagt
     1741 ttcatttgat gctcgatgag tttttctaat cagaattggt taattggttg taacactggc
     1801 agagcattac gctgacttga cgggacggcg caagctcatg accaaaatcc cttaacgtga
     1861 gttacgcgtc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag
     1921 atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg
     1981 tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca
     2041 gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac cacttcaaga
     2101 actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca
     2161 gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc
     2221 agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca
     2281 ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa
     2341 aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc
     2401 cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc
     2461 gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg
     2521 cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt cctgcgttat
     2581 cccctgattc tgtggataac cgtattaccg cctttgagtg agctgatacc gctcgccgca
     2641 gccgaacgac cgagcgcagc gagtc
//
