LOCUS       sequence-101428-        3689 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  MuLE (Multiple Lentiviral Expression) Entry vector containing a SV40
            promoter and EGFP module. Compatible with MultiSite Gateway cloning.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3689)
  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 3689)
  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 eGFP L5-L2
FEATURES             Location/Qualifiers
     source          1..3689
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    complement(23..122)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(152..171)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(153..171)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(176..192)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(176..192)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(294..313)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             305..1114
                     /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(372..391)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      1261..1849
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1750..1769
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2003..2020
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(2179..2206)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(2298..2384)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     2433..2455
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     2447..2464
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     2448..2464
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    2481..2576
                     /gene="mutant version of attL"
                     /label=attL5
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2581..2601)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2586..2915
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2766..2901
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     regulatory      2944..2953
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(2950..2952,2953..2955,2956..3669)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(2950..2952,2953..2955,2956..3666)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(2995..3016)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3256..3275)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3603..3624
                     /label=EGFP-C
                     /note="EGFP, forward primer"
ORIGIN
        1 tcgagcatgc atctagatta ctacccagct ttcttgtaca aagttggcat tataagaaag
       61 cattgcttat caatttgttg caacgaacag gtcactatca gtcaaaataa aatcattatt
      121 tgccatccag ctgcagggcg gccgcgatat cccctatagt gagtcgtatt acatggtcat
      181 agctgtttcc tggcagctct ggcccgtgtc tcaaaatctc tgatgttaca ttgcacaaga
      241 taaaaatata tcatcatgaa caataaaact gtctgcttac ataaacagta atacaagggg
      301 tgttatgagc catattcaac gggaaacgtc gaggccgcga ttaaattcca acatggatgc
      361 tgatttatat gggtataaat gggctcgcga taatgtcggg caatcaggtg cgacaatcta
      421 tcgcttgtat gggaagcccg atgcgccaga gttgtttctg aaacatggca aaggtagcgt
      481 tgccaatgat gttacagatg agatggtcag actaaactgg ctgacggaat ttatgcctct
      541 tccgaccatc aagcatttta tccgtactcc tgatgatgca tggttactca ccactgcgat
      601 ccccggaaaa acagcattcc aggtattaga agaatatcct gattcaggtg aaaatattgt
      661 tgatgcgctg gcagtgttcc tgcgccggtt gcattcgatt cctgtttgta attgtccttt
      721 taacagcgat cgcgtatttc gtctcgctca ggcgcaatca cgaatgaata acggtttggt
      781 tgatgcgagt gattttgatg acgagcgtaa tggctggcct gttgaacaag tctggaaaga
      841 aatgcataaa cttttgccat tctcaccgga ttcagtcgtc actcatggtg atttctcact
      901 tgataacctt atttttgacg aggggaaatt aataggttgt attgatgttg gacgagtcgg
      961 aatcgcagac cgataccagg atcttgccat cctatggaac tgcctcggtg agttttctcc
     1021 ttcattacag aaacggcttt ttcaaaaata tggtattgat aatcctgata tgaataaatt
     1081 gcagtttcat ttgatgctcg atgagttttt ctaatcagaa ttggttaatt ggttgtaaca
     1141 ctggcagagc attacgctga cttgacggga cggcgcaagc tcatgaccaa aatcccttaa
     1201 cgtgagttac gcgtcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc
     1261 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
     1321 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
     1381 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
     1441 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
     1501 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
     1561 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
     1621 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
     1681 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
     1741 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
     1801 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
     1861 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc
     1921 gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg
     1981 ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcccaat
     2041 acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc acgacaggtt
     2101 tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat acgcgtaccg ctagccagga
     2161 agagtttgta gaaacgcaaa aaggccatcc gtcaggatgg ccttctgctt agtttgatgc
     2221 ctggcagttt atggcgggcg tcctgcccgc caccctccgg gccgttgctt cacaacgttc
     2281 aaatccgctc ccggcggatt tgtcctactc aggagagcgt tcaccgacaa acaacagata
     2341 aaacgaaagg cccagtcttc cgactgagcc tttcgtttta tttgatgcct ggcagttccc
     2401 tactctcgcg ttaacgctag catggatgtt ttcccagtca cgacgttgta aaacgacggc
     2461 cagtcttaag ctcgggcccc aaataatgat tttattttga ctgatagtga cctgttcgtt
     2521 gcaacaaatt gatgagcaat gcttttttat aatgccaact ttgtatacaa aagttgctgt
     2581 ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc
     2641 aaagcatgca tctcaattag tcagcaacca ggtgtggaaa gtccccaggc tccccagcag
     2701 gcagaagtat gcaaagcatg catctcaatt agtcagcaac catagtcccg cccctaactc
     2761 cgcccatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat ggctgactaa
     2821 ttttttttat ttatgcagag gccgaggccg cctcggcctc tgagctattc cagaagtagt
     2881 gaggaggctt ttttggaggc ctaggctttt gcaaaaagct tggtaccgag ctcggatcct
     2941 aaagccacca tggtgagcaa gggcgaggag ctgttcaccg gggtggtgcc catcctggtc
     3001 gagctggacg gcgacgtaaa cggccacaag ttcagcgtgt ccggcgaggg cgagggcgat
     3061 gccacctacg gcaagctgac cctgaagttc atctgcacca ccggcaagct gcccgtgccc
     3121 tggcccaccc tcgtgaccac cctgacctac ggcgtgcagt gcttcagccg ctaccccgac
     3181 cacatgaagc agcacgactt cttcaagtcc gccatgcccg aaggctacgt ccaggagcgc
     3241 accatcttct tcaaggacga cggcaactac aagacccgcg ccgaggtgaa gttcgagggc
     3301 gacaccctgg tgaaccgcat cgagctgaag ggcatcgact tcaaggagga cggcaacatc
     3361 ctggggcaca agctggagta caactacaac agccacaacg tctatatcat ggccgacaag
     3421 cagaagaacg gcatcaaggt gaacttcaag atccgccaca acatcgagga cggcagcgtg
     3481 cagctcgccg accactacca gcagaacacc cccatcggcg acggccccgt gctgctgccc
     3541 gacaaccact acctgagcac ccagtccgcc ctgagcaaag accccaacga gaagcgcgat
     3601 cacatggtcc tgctggagtt cgtgaccgcc gccgggatca ctctcggcat ggacgagctg
     3661 tacaagtaaa gcggccgcga ctctagagc
//
