LOCUS       sequence-101454-        4649 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  MuLE (Multiple Lentiviral Expression) Entry vector containing a SV40
            promoter and firefly luciferase module. Compatible with MultiSite 
            Gateway cloning.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4649)
  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 4649)
  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 Luc2 (FF-Luciferase) L5-L2
FEATURES             Location/Qualifiers
     source          1..4649
                     /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"
     regulatory      2986..2995
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2992..4644
                     /codon_start=1
                     /product="firefly luciferase"
                     /label=luciferase
                     /note="synthetic luc2 version of the luciferase gene"
                     /translation="MEDAKNIKKGPAPFYPLEDGTAGEQLHKAMKRYALVPGTIAFTDA
                     HIEVDITYAEYFEMSVRLAEAMKRYGLNTNHRIVVCSENSLQFFMPVLGALFIGVAVAP
                     ANDIYNERELLNSMGISQPTVVFVSKKGLQKILNVQKKLPIIQKIIIMDSKTDYQGFQS
                     MYTFVTSHLPPGFNEYDFVPESFDRDKTIALIMNSSGSTGLPKGVALPHRTACVRFSHA
                     RDPIFGNQIIPDTAILSVVPFHHGFGMFTTLGYLICGFRVVLMYRFEEELFLRSLQDYK
                     IQSALLVPTLFSFFAKSTLIDKYDLSNLHEIASGGAPLSKEVGEAVAKRFHLPGIRQGY
                     GLTETTSAILITPEGDDKPGAVGKVVPFFEAKVVDLDTGKTLGVNQRGELCVRGPMIMS
                     GYVNNPEATNALIDKDGWLHSGDIAYWDEDEHFFIVDRLKSLIKYKGYQVAPAELESIL
                     LQHPNIFDAGVAGLPDDDAGELPAAVVVLEHGKTMTEKEIVDYVASQVTTAKKLRGGVV
                     FVDEVPKGLTGKLDARKIREILIKAKKGGKIAV"
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 aaaaagcttg gtaccgagct cggatccact agtaacggcc
     2941 gccagtgtgc tggaattcgt taacagatcg ggggatccga tatccgccac catggaagat
     3001 gccaaaaaca ttaagaaggg cccagcgcca ttctacccac tcgaagacgg gaccgccggc
     3061 gagcagctgc acaaagccat gaagcgctac gccctggtgc ccggcaccat cgcctttacc
     3121 gacgcacata tcgaggtgga cattacctac gccgagtact tcgagatgag cgttcggctg
     3181 gcagaagcta tgaagcgcta tgggctgaat acaaaccatc ggatcgtggt gtgcagcgag
     3241 aatagcttgc agttcttcat gcccgtgttg ggtgccctgt tcatcggtgt ggctgtggcc
     3301 ccagctaacg acatctacaa cgagcgcgag ctgctgaaca gcatgggcat cagccagccc
     3361 accgtcgtat tcgtgagcaa gaaagggctg caaaagatcc tcaacgtgca aaagaagcta
     3421 ccgatcatac aaaagatcat catcatggat agcaagaccg actaccaggg cttccaaagc
     3481 atgtacacct tcgtgacttc ccatttgcca cccggcttca acgagtacga cttcgtgccc
     3541 gagagcttcg accgggacaa aaccatcgcc ctgatcatga acagtagtgg cagtaccgga
     3601 ttgcccaagg gcgtagccct accgcaccgc accgcttgtg tccgattcag tcatgcccgc
     3661 gaccccatct tcggcaacca gatcatcccc gacaccgcta tcctcagcgt ggtgccattt
     3721 caccacggct tcggcatgtt caccacgctg ggctacttga tctgcggctt tcgggtcgtg
     3781 ctcatgtacc gcttcgagga ggagctattc ttgcgcagct tgcaagacta taagattcaa
     3841 tctgccctgc tggtgcccac actatttagc ttcttcgcta agagcactct catcgacaag
     3901 tacgacctaa gcaacttgca cgagatcgcc agcggcgggg cgccgctcag caaggaggta
     3961 ggtgaggccg tggccaaacg cttccaccta ccaggcatcc gccagggcta cggcctgaca
     4021 gaaacaacca gcgccattct gatcaccccc gaaggggacg acaagcctgg cgcagtaggc
     4081 aaggtggtgc ccttcttcga ggctaaggtg gtggacttgg acaccggtaa gacactgggt
     4141 gtgaaccagc gcggcgagct gtgcgtccgt ggccccatga tcatgagcgg ctacgttaac
     4201 aaccccgagg ctacaaacgc tctcatcgac aaggacggct ggctgcacag cggcgacatc
     4261 gcctactggg acgaggacga gcacttcttc atcgtggacc ggctgaagag cctgatcaaa
     4321 tacaagggct accaggtagc cccagccgaa ctggagagca tcctgctgca acaccccaac
     4381 atcttcgacg ccggggtcgc cggcctgccc gacgacgatg ccggcgagct gcccgccgca
     4441 gtcgtcgtgc tggaacacgg taaaaccatg accgagaagg agatcgtgga ctatgtggcc
     4501 agccaggtta caaccgccaa gaagctgcgc ggtggtgttg tgttcgtgga cgaggtgcct
     4561 aaaggactga ccggcaagtt ggacgcccgc aagatccgcg agattctcat taaggccaag
     4621 aagggcggca agatcgccgt gtaataatt
//
