LOCUS       sequence-101400-        3123 bp    DNA     circular SYN 15-APR-2025
DEFINITION  MuLE (Multiple Lentiviral Expression) Entry vector containing a U6 
            promoter for shRNA expression. Compatible with MultiSite Gateway 
            cloning.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3123)
  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 3123)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 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 U6 L1-R5
FEATURES             Location/Qualifiers
     source          1..3123
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    229..352
                     /gene="mutant version of attR"
                     /label=attR5
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(407..426)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(408..426)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(431..447)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(431..447)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(549..568)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             560..1369
                     /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(627..646)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      1516..2104
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2005..2024
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2258..2275
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(2434..2461)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(2553..2639)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     2688..2710
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     2702..2719
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     2703..2719
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    2735..2834
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        2877..3117
                     /label=U6 promoter
                     /note="Truncated RNA polymerase III promoter for human U6 
                     snRNA used in pSIREN vectors such as 
                     pSIREN-RetroQ-ZsGreen1. Expected transcription start site 
                     is 8bp downstream of this feature."
     primer_bind     2877..2897
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     3048..3067
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
ORIGIN
        1 gatctcggcg ttgtgcatgt cgggcgactg ctccatgatc ttgcgccgct cgatctgcga
       61 ccacaccata aaggcgttca tgggccgctt gatgtggcca ctgggcgtct tgcaccagct
      121 ggggtcgtcc gccttgccgc ccgtcgacgc ggtggagcca ggcgtcgggg aggacgcgat
      181 gcccagctcc aggccggcgc ccgagtccga gctctttttt gaattcgaca actttgtata
      241 caaaagttga acgagaaacg taaaatgata taaatatcaa tatattaaat tagattttgc
      301 ataaaaaaca gactacataa tactgtaaaa cacaacatat ccagtcacta tgaatcaact
      361 acttagatgg tattagtgac ctgtactgca gggcggccgc gatatcccct atagtgagtc
      421 gtattacatg gtcatagctg tttcctggca gctctggccc gtgtctcaaa atctctgatg
      481 ttacattgca caagataaaa atatatcatc atgaacaata aaactgtctg cttacataaa
      541 cagtaataca aggggtgtta tgagccatat tcaacgggaa acgtcgaggc cgcgattaaa
      601 ttccaacatg gatgctgatt tatatgggta taaatgggct cgcgataatg tcgggcaatc
      661 aggtgcgaca atctatcgct tgtatgggaa gcccgatgcg ccagagttgt ttctgaaaca
      721 tggcaaaggt agcgttgcca atgatgttac agatgagatg gtcagactaa actggctgac
      781 ggaatttatg cctcttccga ccatcaagca ttttatccgt actcctgatg atgcatggtt
      841 actcaccact gcgatccccg gaaaaacagc attccaggta ttagaagaat atcctgattc
      901 aggtgaaaat attgttgatg cgctggcagt gttcctgcgc cggttgcatt cgattcctgt
      961 ttgtaattgt ccttttaaca gcgatcgcgt atttcgtctc gctcaggcgc aatcacgaat
     1021 gaataacggt ttggttgatg cgagtgattt tgatgacgag cgtaatggct ggcctgttga
     1081 acaagtctgg aaagaaatgc ataaactttt gccattctca ccggattcag tcgtcactca
     1141 tggtgatttc tcacttgata accttatttt tgacgagggg aaattaatag gttgtattga
     1201 tgttggacga gtcggaatcg cagaccgata ccaggatctt gccatcctat ggaactgcct
     1261 cggtgagttt tctccttcat tacagaaacg gctttttcaa aaatatggta ttgataatcc
     1321 tgatatgaat aaattgcagt ttcatttgat gctcgatgag tttttctaat cagaattggt
     1381 taattggttg taacactggc agagcattac gctgacttga cgggacggcg caagctcatg
     1441 accaaaatcc cttaacgtga gttacgcgtc gttccactga gcgtcagacc ccgtagaaaa
     1501 gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa
     1561 aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc
     1621 gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta
     1681 gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct
     1741 gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg
     1801 atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag
     1861 cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc
     1921 cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg
     1981 agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt
     2041 tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg
     2101 gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca
     2161 catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg cctttgagtg
     2221 agctgatacc gctcgccgca gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc
     2281 ggaagagcgc ccaatacgca aaccgcctct ccccgcgcgt tggccgattc attaatgcag
     2341 ctggcacgac aggtttcccg actggaaagc gggcagtgag cgcaacgcaa ttaatacgcg
     2401 taccgctagc caggaagagt ttgtagaaac gcaaaaaggc catccgtcag gatggccttc
     2461 tgcttagttt gatgcctggc agtttatggc gggcgtcctg cccgccaccc tccgggccgt
     2521 tgcttcacaa cgttcaaatc cgctcccggc ggatttgtcc tactcaggag agcgttcacc
     2581 gacaaacaac agataaaacg aaaggcccag tcttccgact gagcctttcg ttttatttga
     2641 tgcctggcag ttccctactc tcgcgttaac gctagcatgg atgttttccc agtcacgacg
     2701 ttgtaaaacg acggccagtc ttaagctcgg gccccaaata atgattttat tttgactgat
     2761 agtgacctgt tcgttgcaac aaattgatga gcaatgcttt tttataatgc caactttgta
     2821 caaaaaagca ggctctatcg atccgagact agcctcgagc ggccgccccc ttcaccgagg
     2881 gcctatttcc catgattcct tcatatttgc atatacgata caaggctgtt agagagataa
     2941 ttggaattaa tttgactgta aacacaaaga tattagtaca aaatacgtga cgtagaaagt
     3001 aataatttct tgggtagttt gcagttttaa aattatgttt taaaatggac tatcatatgc
     3061 ttaccgtaac ttgaaagtat ttcgatttct tggctttata tatcttgtgg aaaggacgaa
     3121 aca
//
