LOCUS       sequence-101385-        2994 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  MuLE (Multiple Lentiviral Expression) Entry vector containing a SV40
            promoter and multiple cloning site module. Compatible with MultiSite
            Gateway cloning.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2994)
  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 2994)
  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 L3-L2
FEATURES             Location/Qualifiers
     source          1..2994
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    complement(101..200)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(230..249)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(231..249)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(254..270)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(254..270)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(372..391)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             383..1192
                     /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(450..469)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      1339..1927
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1828..1847
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2081..2098
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(2257..2284)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(2376..2462)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     2511..2533
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     2525..2542
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     2526..2542
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    2559..2654
                     /gene="mutant version of attL"
                     /label=attL3
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2659..2679)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2664..2993
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2844..2979
                     /label=SV40 ori
                     /note="SV40 origin of replication"
ORIGIN
        1 agcttggtac cgagctcgga tccactagta acggccgcca gtgtgctgga attctgcaga
       61 tatccatcac actggcggcc gctcgagcat gcatctagat acccagcttt cttgtacaaa
      121 gttggcatta taagaaagca ttgcttatca atttgttgca acgaacaggt cactatcagt
      181 caaaataaaa tcattatttg ccatccagct gcagggcggc cgcgatatcc cctatagtga
      241 gtcgtattac atggtcatag ctgtttcctg gcagctctgg cccgtgtctc aaaatctctg
      301 atgttacatt gcacaagata aaaatatatc atcatgaaca ataaaactgt ctgcttacat
      361 aaacagtaat acaaggggtg ttatgagcca tattcaacgg gaaacgtcga ggccgcgatt
      421 aaattccaac atggatgctg atttatatgg gtataaatgg gctcgcgata atgtcgggca
      481 atcaggtgcg acaatctatc gcttgtatgg gaagcccgat gcgccagagt tgtttctgaa
      541 acatggcaaa ggtagcgttg ccaatgatgt tacagatgag atggtcagac taaactggct
      601 gacggaattt atgcctcttc cgaccatcaa gcattttatc cgtactcctg atgatgcatg
      661 gttactcacc actgcgatcc ccggaaaaac agcattccag gtattagaag aatatcctga
      721 ttcaggtgaa aatattgttg atgcgctggc agtgttcctg cgccggttgc attcgattcc
      781 tgtttgtaat tgtcctttta acagcgatcg cgtatttcgt ctcgctcagg cgcaatcacg
      841 aatgaataac ggtttggttg atgcgagtga ttttgatgac gagcgtaatg gctggcctgt
      901 tgaacaagtc tggaaagaaa tgcataaact tttgccattc tcaccggatt cagtcgtcac
      961 tcatggtgat ttctcacttg ataaccttat ttttgacgag gggaaattaa taggttgtat
     1021 tgatgttgga cgagtcggaa tcgcagaccg ataccaggat cttgccatcc tatggaactg
     1081 cctcggtgag ttttctcctt cattacagaa acggcttttt caaaaatatg gtattgataa
     1141 tcctgatatg aataaattgc agtttcattt gatgctcgat gagtttttct aatcagaatt
     1201 ggttaattgg ttgtaacact ggcagagcat tacgctgact tgacgggacg gcgcaagctc
     1261 atgaccaaaa tcccttaacg tgagttacgc gtcgttccac tgagcgtcag accccgtaga
     1321 aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac
     1381 aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt
     1441 tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc
     1501 gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat
     1561 cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag
     1621 acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc
     1681 cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag
     1741 cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac
     1801 aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg
     1861 gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct
     1921 atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc
     1981 tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgcctttga
     2041 gtgagctgat accgctcgcc gcagccgaac gaccgagcgc agcgagtcag tgagcgagga
     2101 agcggaagag cgcccaatac gcaaaccgcc tctccccgcg cgttggccga ttcattaatg
     2161 cagctggcac gacaggtttc ccgactggaa agcgggcagt gagcgcaacg caattaatac
     2221 gcgtaccgct agccaggaag agtttgtaga aacgcaaaaa ggccatccgt caggatggcc
     2281 ttctgcttag tttgatgcct ggcagtttat ggcgggcgtc ctgcccgcca ccctccgggc
     2341 cgttgcttca caacgttcaa atccgctccc ggcggatttg tcctactcag gagagcgttc
     2401 accgacaaac aacagataaa acgaaaggcc cagtcttccg actgagcctt tcgttttatt
     2461 tgatgcctgg cagttcccta ctctcgcgtt aacgctagca tggatgtttt cccagtcacg
     2521 acgttgtaaa acgacggcca gtcttaagct cgggccccaa ataatgattt tattttgact
     2581 gatagtgacc tgttcgttgc aacaaattga tgagcaatgc ttttttataa tgccaacttt
     2641 gtataataaa gttgctgtgg aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc
     2701 cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccagg tgtggaaagt
     2761 ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca
     2821 tagtcccgcc cctaactccg cccatcccgc ccctaactcc gcccagttcc gcccattctc
     2881 cgccccatgg ctgactaatt ttttttattt atgcagaggc cgaggccgcc tcggcctctg
     2941 agctattcca gaagtagtga ggaggctttt ttggaggcct aggcttttgc aaaa
//
