LOCUS       sequence-347562-        2668 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 2668)
  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 2668)
  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 L1-L4
FEATURES             Location/Qualifiers
     source          1..2668
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      complement(137..164)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(256..342)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     391..413
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     405..422
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     406..422
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    438..537
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     protein_bind    complement(652..747)
                     /gene="mutant version of attL"
                     /label=attL4
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(778..797)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(779..797)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(802..818)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(802..818)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(920..939)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             931..1740
                     /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(998..1017)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      1887..2475
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2376..2395
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2629..2646
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 gcaaaccgcc tctccccgcg cgttggccga ttcattaatg cagctggcac gacaggtttc
       61 ccgactggaa agcgggcagt gagcgcaacg caattaatac gcgtaccgct agccaggaag
      121 agtttgtaga aacgcaaaaa ggccatccgt caggatggcc ttctgcttag tttgatgcct
      181 ggcagtttat ggcgggcgtc ctgcccgcca ccctccgggc cgttgcttca caacgttcaa
      241 atccgctccc ggcggatttg tcctactcag gagagcgttc accgacaaac aacagataaa
      301 acgaaaggcc cagtcttccg actgagcctt tcgttttatt tgatgcctgg cagttcccta
      361 ctctcgcgtt aacgctagca tggatgtttt cccagtcacg acgttgtaaa acgacggcca
      421 gtcttaagct cgggccccaa ataatgattt tattttgact gatagtgacc tgttcgttgc
      481 aacaaattga tgagcaatgc ttttttataa tgccaacttt gtacaaaaaa gcaggctgga
      541 gacccaagct tggtaccgag ctcggatcca ctagtaacgg ccgccagtgt gctggaattc
      601 tgcagatatc catcacactg gcggccgctc gagcatgcat ctagaggcac ccaacttttc
      661 tatacaaagt tggcattata agaaagcatt gcttatcaat ttgttgcaac gaacaggtca
      721 ctatcagtca aaataaaatc attatttgcc atccagctgc agggcggccg cgatatcccc
      781 tatagtgagt cgtattacat ggtcatagct gtttcctggc agctctggcc cgtgtctcaa
      841 aatctctgat gttacattgc acaagataaa aatatatcat catgaacaat aaaactgtct
      901 gcttacataa acagtaatac aaggggtgtt atgagccata ttcaacggga aacgtcgagg
      961 ccgcgattaa attccaacat ggatgctgat ttatatgggt ataaatgggc tcgcgataat
     1021 gtcgggcaat caggtgcgac aatctatcgc ttgtatggga agcccgatgc gccagagttg
     1081 tttctgaaac atggcaaagg tagcgttgcc aatgatgtta cagatgagat ggtcagacta
     1141 aactggctga cggaatttat gcctcttccg accatcaagc attttatccg tactcctgat
     1201 gatgcatggt tactcaccac tgcgatcccc ggaaaaacag cattccaggt attagaagaa
     1261 tatcctgatt caggtgaaaa tattgttgat gcgctggcag tgttcctgcg ccggttgcat
     1321 tcgattcctg tttgtaattg tccttttaac agcgatcgcg tatttcgtct cgctcaggcg
     1381 caatcacgaa tgaataacgg tttggttgat gcgagtgatt ttgatgacga gcgtaatggc
     1441 tggcctgttg aacaagtctg gaaagaaatg cataaacttt tgccattctc accggattca
     1501 gtcgtcactc atggtgattt ctcacttgat aaccttattt ttgacgaggg gaaattaata
     1561 ggttgtattg atgttggacg agtcggaatc gcagaccgat accaggatct tgccatccta
     1621 tggaactgcc tcggtgagtt ttctccttca ttacagaaac ggctttttca aaaatatggt
     1681 attgataatc ctgatatgaa taaattgcag tttcatttga tgctcgatga gtttttctaa
     1741 tcagaattgg ttaattggtt gtaacactgg cagagcatta cgctgacttg acgggacggc
     1801 gcaagctcat gaccaaaatc ccttaacgtg agttacgcgt cgttccactg agcgtcagac
     1861 cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc
     1921 ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca
     1981 actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta
     2041 gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct
     2101 ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg
     2161 gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc
     2221 acacagccca gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta
     2281 tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg
     2341 gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt
     2401 cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg
     2461 cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg
     2521 ccttttgctc acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc
     2581 gcctttgagt gagctgatac cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg
     2641 agcgaggaag cggaagagcg cccaatac
//
