LOCUS       sequence-101449-        3585 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  MuLE (Multiple Lentiviral Expression) Entry vector containing a SV40
            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 3585)
  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 3585)
  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 SV40 PuroR L5-L2
FEATURES             Location/Qualifiers
     source          1..3585
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    complement(11..110)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(140..159)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(141..159)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(164..180)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(164..180)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(282..301)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             293..1102
                     /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(360..379)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      1249..1837
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1738..1757
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1991..2008
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(2167..2194)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(2286..2372)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     2421..2443
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     2435..2452
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     2436..2452
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    2469..2564
                     /gene="mutant version of attL"
                     /label=attL5
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2569..2589)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2574..2903
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2754..2889
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             2913..3512
                     /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(2913..2932)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     3409..3429
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
ORIGIN
        1 ctagattact acccagcttt cttgtacaaa gttggcatta taagaaagca ttgcttatca
       61 atttgttgca acgaacaggt cactatcagt caaaataaaa tcattatttg ccatccagct
      121 gcagggcggc cgcgatatcc cctatagtga gtcgtattac atggtcatag ctgtttcctg
      181 gcagctctgg cccgtgtctc aaaatctctg atgttacatt gcacaagata aaaatatatc
      241 atcatgaaca ataaaactgt ctgcttacat aaacagtaat acaaggggtg ttatgagcca
      301 tattcaacgg gaaacgtcga ggccgcgatt aaattccaac atggatgctg atttatatgg
      361 gtataaatgg gctcgcgata atgtcgggca atcaggtgcg acaatctatc gcttgtatgg
      421 gaagcccgat gcgccagagt tgtttctgaa acatggcaaa ggtagcgttg ccaatgatgt
      481 tacagatgag atggtcagac taaactggct gacggaattt atgcctcttc cgaccatcaa
      541 gcattttatc cgtactcctg atgatgcatg gttactcacc actgcgatcc ccggaaaaac
      601 agcattccag gtattagaag aatatcctga ttcaggtgaa aatattgttg atgcgctggc
      661 agtgttcctg cgccggttgc attcgattcc tgtttgtaat tgtcctttta acagcgatcg
      721 cgtatttcgt ctcgctcagg cgcaatcacg aatgaataac ggtttggttg atgcgagtga
      781 ttttgatgac gagcgtaatg gctggcctgt tgaacaagtc tggaaagaaa tgcataaact
      841 tttgccattc tcaccggatt cagtcgtcac tcatggtgat ttctcacttg ataaccttat
      901 ttttgacgag gggaaattaa taggttgtat tgatgttgga cgagtcggaa tcgcagaccg
      961 ataccaggat cttgccatcc tatggaactg cctcggtgag ttttctcctt cattacagaa
     1021 acggcttttt caaaaatatg gtattgataa tcctgatatg aataaattgc agtttcattt
     1081 gatgctcgat gagtttttct aatcagaatt ggttaattgg ttgtaacact ggcagagcat
     1141 tacgctgact tgacgggacg gcgcaagctc atgaccaaaa tcccttaacg tgagttacgc
     1201 gtcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt
     1261 ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg
     1321 tttgccggat caagagctac caactctttt tccgaaggta actggcttca gcagagcgca
     1381 gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt
     1441 agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga
     1501 taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc
     1561 gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact
     1621 gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga
     1681 caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg
     1741 aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt
     1801 tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt
     1861 acggttcctg gccttttgct ggccttttgc tcacatgttc tttcctgcgt tatcccctga
     1921 ttctgtggat aaccgtatta ccgcctttga gtgagctgat accgctcgcc gcagccgaac
     1981 gaccgagcgc agcgagtcag tgagcgagga agcggaagag cgcccaatac gcaaaccgcc
     2041 tctccccgcg cgttggccga ttcattaatg cagctggcac gacaggtttc ccgactggaa
     2101 agcgggcagt gagcgcaacg caattaatac gcgtaccgct agccaggaag agtttgtaga
     2161 aacgcaaaaa ggccatccgt caggatggcc ttctgcttag tttgatgcct ggcagtttat
     2221 ggcgggcgtc ctgcccgcca ccctccgggc cgttgcttca caacgttcaa atccgctccc
     2281 ggcggatttg tcctactcag gagagcgttc accgacaaac aacagataaa acgaaaggcc
     2341 cagtcttccg actgagcctt tcgttttatt tgatgcctgg cagttcccta ctctcgcgtt
     2401 aacgctagca tggatgtttt cccagtcacg acgttgtaaa acgacggcca gtcttaagct
     2461 cgggccccaa ataatgattt tattttgact gatagtgacc tgttcgttgc aacaaattga
     2521 tgagcaatgc ttttttataa tgccaacttt gtatacaaaa gttgctgtgg aatgtgtgtc
     2581 agttagggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc
     2641 tcaattagtc agcaaccagg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc
     2701 aaagcatgca tctcaattag tcagcaacca tagtcccgcc cctaactccg cccatcccgc
     2761 ccctaactcc gcccagttcc gcccattctc cgccccatgg ctgactaatt ttttttattt
     2821 atgcagaggc cgaggccgcc tcggcctctg agctattcca gaagtagtga ggaggctttt
     2881 ttggaggcct aggcttttgc aaaaagctta ccatgaccga gtacaagccc acggtgcgcc
     2941 tcgccacccg cgacgacgtc cccagggccg tacgcaccct cgccgccgcg ttcgccgact
     3001 accccgccac gcgccacacc gtcgatccgg accgccacat cgagcgggtc accgagctgc
     3061 aagaactctt cctcacgcgc gtcgggctcg acatcggcaa ggtgtgggtc gcggacgacg
     3121 gcgccgcggt ggcggtctgg accacgccgg agagcgtcga agcgggggcg gtgttcgccg
     3181 agatcggccc gcgcatggcc gagttgagcg gttcccggct ggccgcgcag caacagatgg
     3241 aaggcctcct ggcgccgcac cggcccaagg agcccgcgtg gttcctggcc accgtcggcg
     3301 tctcgcccga ccaccagggc aagggtctgg gcagcgccgt cgtgctcccc ggagtggagg
     3361 cggccgagcg cgccggggtg cccgccttcc tggagacctc cgcgccccgc aacctcccct
     3421 tctacgagcg gctcggcttc accgtcaccg ccgacgtcga ggtgcccgaa ggaccgcgca
     3481 cctggtgcat gacccgcaag cccggtgcct gacgcccgcc ccacgacccg cagcgcccga
     3541 ccgaaaggag cgcacgaccc catgcatcgc cagtcgagca tgcat
//
