LOCUS       sequence-101429-        3685 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 3685)
  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 3685)
  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 L3-L2
FEATURES             Location/Qualifiers
     source          1..3685
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    complement(19..118)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(148..167)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(149..167)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(172..188)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(172..188)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(290..309)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             301..1110
                     /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(368..387)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      1257..1845
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1746..1765
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1999..2016
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(2175..2202)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(2294..2380)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     2429..2451
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     2443..2460
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     2444..2460
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    2477..2572
                     /gene="mutant version of attL"
                     /label=attL3
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2577..2597)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2582..2911
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2762..2897
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     regulatory      2940..2949
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(2946..2948,2949..2951,2952..3665)
                     /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(2946..2948,2949..2951,2952..3662)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(2991..3012)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3252..3271)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3599..3620
                     /label=EGFP-C
                     /note="EGFP, forward primer"
ORIGIN
        1 tcgagcatgc atctagatac ccagctttct tgtacaaagt tggcattata agaaagcatt
       61 gcttatcaat ttgttgcaac gaacaggtca ctatcagtca aaataaaatc attatttgcc
      121 atccagctgc agggcggccg cgatatcccc tatagtgagt cgtattacat ggtcatagct
      181 gtttcctggc agctctggcc cgtgtctcaa aatctctgat gttacattgc acaagataaa
      241 aatatatcat catgaacaat aaaactgtct gcttacataa acagtaatac aaggggtgtt
      301 atgagccata ttcaacggga aacgtcgagg ccgcgattaa attccaacat ggatgctgat
      361 ttatatgggt ataaatgggc tcgcgataat gtcgggcaat caggtgcgac aatctatcgc
      421 ttgtatggga agcccgatgc gccagagttg tttctgaaac atggcaaagg tagcgttgcc
      481 aatgatgtta cagatgagat ggtcagacta aactggctga cggaatttat gcctcttccg
      541 accatcaagc attttatccg tactcctgat gatgcatggt tactcaccac tgcgatcccc
      601 ggaaaaacag cattccaggt attagaagaa tatcctgatt caggtgaaaa tattgttgat
      661 gcgctggcag tgttcctgcg ccggttgcat tcgattcctg tttgtaattg tccttttaac
      721 agcgatcgcg tatttcgtct cgctcaggcg caatcacgaa tgaataacgg tttggttgat
      781 gcgagtgatt ttgatgacga gcgtaatggc tggcctgttg aacaagtctg gaaagaaatg
      841 cataaacttt tgccattctc accggattca gtcgtcactc atggtgattt ctcacttgat
      901 aaccttattt ttgacgaggg gaaattaata ggttgtattg atgttggacg agtcggaatc
      961 gcagaccgat accaggatct tgccatccta tggaactgcc tcggtgagtt ttctccttca
     1021 ttacagaaac ggctttttca aaaatatggt attgataatc ctgatatgaa taaattgcag
     1081 tttcatttga tgctcgatga gtttttctaa tcagaattgg ttaattggtt gtaacactgg
     1141 cagagcatta cgctgacttg acgggacggc gcaagctcat gaccaaaatc ccttaacgtg
     1201 agttacgcgt cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga
     1261 gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg
     1321 gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc
     1381 agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca ccacttcaag
     1441 aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc
     1501 agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg
     1561 cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac
     1621 accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga
     1681 aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt
     1741 ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag
     1801 cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg
     1861 gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta
     1921 tcccctgatt ctgtggataa ccgtattacc gcctttgagt gagctgatac cgctcgccgc
     1981 agccgaacga ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg cccaatacgc
     2041 aaaccgcctc tccccgcgcg ttggccgatt cattaatgca gctggcacga caggtttccc
     2101 gactggaaag cgggcagtga gcgcaacgca attaatacgc gtaccgctag ccaggaagag
     2161 tttgtagaaa cgcaaaaagg ccatccgtca ggatggcctt ctgcttagtt tgatgcctgg
     2221 cagtttatgg cgggcgtcct gcccgccacc ctccgggccg ttgcttcaca acgttcaaat
     2281 ccgctcccgg cggatttgtc ctactcagga gagcgttcac cgacaaacaa cagataaaac
     2341 gaaaggccca gtcttccgac tgagcctttc gttttatttg atgcctggca gttccctact
     2401 ctcgcgttaa cgctagcatg gatgttttcc cagtcacgac gttgtaaaac gacggccagt
     2461 cttaagctcg ggccccaaat aatgatttta ttttgactga tagtgacctg ttcgttgcaa
     2521 caaattgatg agcaatgctt ttttataatg ccaactttgt ataataaagt tgctgtggaa
     2581 tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag
     2641 catgcatctc aattagtcag caaccaggtg tggaaagtcc ccaggctccc cagcaggcag
     2701 aagtatgcaa agcatgcatc tcaattagtc agcaaccata gtcccgcccc taactccgcc
     2761 catcccgccc ctaactccgc ccagttccgc ccattctccg ccccatggct gactaatttt
     2821 ttttatttat gcagaggccg aggccgcctc ggcctctgag ctattccaga agtagtgagg
     2881 aggctttttt ggaggcctag gcttttgcaa aaagcttggt accgagctcg gatcctaaag
     2941 ccaccatggt gagcaagggc gaggagctgt tcaccggggt ggtgcccatc ctggtcgagc
     3001 tggacggcga cgtaaacggc cacaagttca gcgtgtccgg cgagggcgag ggcgatgcca
     3061 cctacggcaa gctgaccctg aagttcatct gcaccaccgg caagctgccc gtgccctggc
     3121 ccaccctcgt gaccaccctg acctacggcg tgcagtgctt cagccgctac cccgaccaca
     3181 tgaagcagca cgacttcttc aagtccgcca tgcccgaagg ctacgtccag gagcgcacca
     3241 tcttcttcaa ggacgacggc aactacaaga cccgcgccga ggtgaagttc gagggcgaca
     3301 ccctggtgaa ccgcatcgag ctgaagggca tcgacttcaa ggaggacggc aacatcctgg
     3361 ggcacaagct ggagtacaac tacaacagcc acaacgtcta tatcatggcc gacaagcaga
     3421 agaacggcat caaggtgaac ttcaagatcc gccacaacat cgaggacggc agcgtgcagc
     3481 tcgccgacca ctaccagcag aacaccccca tcggcgacgg ccccgtgctg ctgcccgaca
     3541 accactacct gagcacccag tccgccctga gcaaagaccc caacgagaag cgcgatcaca
     3601 tggtcctgct ggagttcgtg accgccgccg ggatcactct cggcatggac gagctgtaca
     3661 agtaaagcgg ccgcgactct agagc
//
