LOCUS       sequence-101447-        3925 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  MuLE (Multiple Lentiviral Expression) Entry vector containing a CMV 
            promoter and Puromycin module. Compatible with MultiSite Gateway 
            cloning.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3925)
  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 3925)
  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 PuroR L3-L2
FEATURES             Location/Qualifiers
     source          1..3925
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    complement(9..108)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        complement(139..157)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(162..178)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(162..178)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(280..299)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             291..1100
                     /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(358..377)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      1247..1835
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1736..1755
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1989..2006
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(2165..2192)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(2284..2370)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     2419..2441
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     2433..2450
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     2434..2450
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    2467..2562
                     /gene="mutant version of attL"
                     /label=attL3
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     enhancer        2590..2969
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2970..3173
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     3124..3144
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     promoter        3218..3236
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             3253..3852
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(3253..3272)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     3749..3769
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
ORIGIN
        1 ctagaggtac ccagctttct tgtacaaagt tggcattata agaaagcatt gcttatcaat
       61 ttgttgcaac gaacaggtca ctatcagtca aaataaaatc attatttgcc atccagctgc
      121 agggcggccg cgatatcccc tatagtgagt cgtattacat ggtcatagct gtttcctggc
      181 agctctggcc cgtgtctcaa aatctctgat gttacattgc acaagataaa aatatatcat
      241 catgaacaat aaaactgtct gcttacataa acagtaatac aaggggtgtt atgagccata
      301 ttcaacggga aacgtcgagg ccgcgattaa attccaacat ggatgctgat ttatatgggt
      361 ataaatgggc tcgcgataat gtcgggcaat caggtgcgac aatctatcgc ttgtatggga
      421 agcccgatgc gccagagttg tttctgaaac atggcaaagg tagcgttgcc aatgatgtta
      481 cagatgagat ggtcagacta aactggctga cggaatttat gcctcttccg accatcaagc
      541 attttatccg tactcctgat gatgcatggt tactcaccac tgcgatcccc ggaaaaacag
      601 cattccaggt attagaagaa tatcctgatt caggtgaaaa tattgttgat gcgctggcag
      661 tgttcctgcg ccggttgcat tcgattcctg tttgtaattg tccttttaac agcgatcgcg
      721 tatttcgtct cgctcaggcg caatcacgaa tgaataacgg tttggttgat gcgagtgatt
      781 ttgatgacga gcgtaatggc tggcctgttg aacaagtctg gaaagaaatg cataaacttt
      841 tgccattctc accggattca gtcgtcactc atggtgattt ctcacttgat aaccttattt
      901 ttgacgaggg gaaattaata ggttgtattg atgttggacg agtcggaatc gcagaccgat
      961 accaggatct tgccatccta tggaactgcc tcggtgagtt ttctccttca ttacagaaac
     1021 ggctttttca aaaatatggt attgataatc ctgatatgaa taaattgcag tttcatttga
     1081 tgctcgatga gtttttctaa tcagaattgg ttaattggtt gtaacactgg cagagcatta
     1141 cgctgacttg acgggacggc gcaagctcat gaccaaaatc ccttaacgtg agttacgcgt
     1201 cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt
     1261 ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt
     1321 tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga
     1381 taccaaatac tgttcttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag
     1441 caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata
     1501 agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg
     1561 gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga
     1621 gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca
     1681 ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa
     1741 acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt
     1801 tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac
     1861 ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta tcccctgatt
     1921 ctgtggataa ccgtattacc gcctttgagt gagctgatac cgctcgccgc agccgaacga
     1981 ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg cccaatacgc aaaccgcctc
     2041 tccccgcgcg ttggccgatt cattaatgca gctggcacga caggtttccc gactggaaag
     2101 cgggcagtga gcgcaacgca attaatacgc gtaccgctag ccaggaagag tttgtagaaa
     2161 cgcaaaaagg ccatccgtca ggatggcctt ctgcttagtt tgatgcctgg cagtttatgg
     2221 cgggcgtcct gcccgccacc ctccgggccg ttgcttcaca acgttcaaat ccgctcccgg
     2281 cggatttgtc ctactcagga gagcgttcac cgacaaacaa cagataaaac gaaaggccca
     2341 gtcttccgac tgagcctttc gttttatttg atgcctggca gttccctact ctcgcgttaa
     2401 cgctagcatg gatgttttcc cagtcacgac gttgtaaaac gacggccagt cttaagctcg
     2461 ggccccaaat aatgatttta ttttgactga tagtgacctg ttcgttgcaa caaattgatg
     2521 agcaatgctt ttttataatg ccaactttgt ataataaagt tggcgatgta cgggccagat
     2581 atacgcgttg acattgatta ttgactagtt attaatagta atcaattacg gggtcattag
     2641 ttcatagccc atatatggag ttccgcgtta cataacttac ggtaaatggc ccgcctggct
     2701 gaccgcccaa cgacccccgc ccattgacgt caataatgac gtatgttccc atagtaacgc
     2761 caatagggac tttccattga cgtcaatggg tggagtattt acggtaaact gcccacttgg
     2821 cagtacatca agtgtatcat atgccaagta cgccccctat tgacgtcaat gacggtaaat
     2881 ggcccgcctg gcattatgcc cagtacatga ccttatggga ctttcctact tggcagtaca
     2941 tctacgtatt agtcatcgct attaccatgg tgatgcggtt ttggcagtac atcaatgggc
     3001 gtggatagcg gtttgactca cggggatttc caagtctcca ccccattgac gtcaatggga
     3061 gtttgttttg gcaccaaaat caacgggact ttccaaaatg tcgtaacaac tccgccccat
     3121 tgacgcaaat gggcggtagg cgtgtacggt gggaggtcta tataagcaga gctctctggc
     3181 taactagaga acccactgct tactggctta tcgaaattaa tacgactcac tatagggaga
     3241 cccaagctta ccatgaccga gtacaagccc acggtgcgcc tcgccacccg cgacgacgtc
     3301 cccagggccg tacgcaccct cgccgccgcg ttcgccgact accccgccac gcgccacacc
     3361 gtcgatccgg accgccacat cgagcgggtc accgagctgc aagaactctt cctcacgcgc
     3421 gtcgggctcg acatcggcaa ggtgtgggtc gcggacgacg gcgccgcggt ggcggtctgg
     3481 accacgccgg agagcgtcga agcgggggcg gtgttcgccg agatcggccc gcgcatggcc
     3541 gagttgagcg gttcccggct ggccgcgcag caacagatgg aaggcctcct ggcgccgcac
     3601 cggcccaagg agcccgcgtg gttcctggcc accgtcggcg tctcgcccga ccaccagggc
     3661 aagggtctgg gcagcgccgt cgtgctcccc ggagtggagg cggccgagcg cgccggggtg
     3721 cccgccttcc tggagacctc cgcgccccgc aacctcccct tctacgagcg gctcggcttc
     3781 accgtcaccg ccgacgtcga ggtgcccgaa ggaccgcgca cctggtgcat gacccgcaag
     3841 cccggtgcct gacgcccgcc ccacgacccg cagcgcccga ccgaaaggag cgcacgaccc
     3901 catgcatcgc cagtcgagca tgcat
//
