LOCUS       Exported                3019 bp ds-DNA     circular SYN 13-MAY-2021
DEFINITION  MuLE (Multiple Lentiviral Expression) Entry vector containing a U6 
            promoter and a non-silencing (scrambled) shRNA module. Compatible 
            with MultiSite Gateway cloning.
ACCESSION   .
VERSION     .
KEYWORDS    pMuLE ENTR U6 n.s shRNA R4-R3
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3019)
  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 3019)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 13, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..3019
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     protein_bind    8..131
                     /gene="mutant version of attR"
                     /label=attR3
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(186..205)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(187..205)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(210..226)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(210..226)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(328..347)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             339..1148
                     /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(406..425)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      1295..1883
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1784..1803
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2037..2054
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      2213..2240
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      2332..2418
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     2467..2489
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     2481..2498
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     2482..2498
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(2548..2672)
                     /gene="mutant version of attR"
                     /label=attR4
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        2714..2954
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA"
     primer_bind     2714..2734
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     2885..2904
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
ORIGIN
        1 aattcgacaa ctttgtataa taaagttgaa cgagaaacgt aaaatgatat aaatatcaat
       61 atattaaatt agattttgca taaaaaacag actacataat actgtaaaac acaacatatc
      121 cagtcactat gaatcaacta cttagatggt attagtgacc tgtactgcag ggcggccgcg
      181 atatccccta tagtgagtcg tattacatgg tcatagctgt ttcctggcag ctctggcccg
      241 tgtctcaaaa tctctgatgt tacattgcac aagataaaaa tatatcatca tgaacaataa
      301 aactgtctgc ttacataaac agtaatacaa ggggtgttat gagccatatt caacgggaaa
      361 cgtcgaggcc gcgattaaat tccaacatgg atgctgattt atatgggtat aaatgggctc
      421 gcgataatgt cgggcaatca ggtgcgacaa tctatcgctt gtatgggaag cccgatgcgc
      481 cagagttgtt tctgaaacat ggcaaaggta gcgttgccaa tgatgttaca gatgagatgg
      541 tcagactaaa ctggctgacg gaatttatgc ctcttccgac catcaagcat tttatccgta
      601 ctcctgatga tgcatggtta ctcaccactg cgatccccgg aaaaacagca ttccaggtat
      661 tagaagaata tcctgattca ggtgaaaata ttgttgatgc gctggcagtg ttcctgcgcc
      721 ggttgcattc gattcctgtt tgtaattgtc cttttaacag cgatcgcgta tttcgtctcg
      781 ctcaggcgca atcacgaatg aataacggtt tggttgatgc gagtgatttt gatgacgagc
      841 gtaatggctg gcctgttgaa caagtctgga aagaaatgca taaacttttg ccattctcac
      901 cggattcagt cgtcactcat ggtgatttct cacttgataa ccttattttt gacgagggga
      961 aattaatagg ttgtattgat gttggacgag tcggaatcgc agaccgatac caggatcttg
     1021 ccatcctatg gaactgcctc ggtgagtttt ctccttcatt acagaaacgg ctttttcaaa
     1081 aatatggtat tgataatcct gatatgaata aattgcagtt tcatttgatg ctcgatgagt
     1141 ttttctaatc agaattggtt aattggttgt aacactggca gagcattacg ctgacttgac
     1201 gggacggcgc aagctcatga ccaaaatccc ttaacgtgag ttacgcgtcg ttccactgag
     1261 cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa
     1321 tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag
     1381 agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg
     1441 ttcttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat
     1501 acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta
     1561 ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg
     1621 gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc
     1681 gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa
     1741 gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc
     1801 tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt
     1861 caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct
     1921 tttgctggcc ttttgctcac atgttctttc ctgcgttatc ccctgattct gtggataacc
     1981 gtattaccgc ctttgagtga gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg
     2041 agtcagtgag cgaggaagcg gaagagcgcc caatacgcaa accgcctctc cccgcgcgtt
     2101 ggccgattca ttaatgcagc tggcacgaca ggtttcccga ctggaaagcg ggcagtgagc
     2161 gcaacgcaat taatacgcgt accgctagcc aggaagagtt tgtagaaacg caaaaaggcc
     2221 atccgtcagg atggccttct gcttagtttg atgcctggca gtttatggcg ggcgtcctgc
     2281 ccgccaccct ccgggccgtt gcttcacaac gttcaaatcc gctcccggcg gatttgtcct
     2341 actcaggaga gcgttcaccg acaaacaaca gataaaacga aaggcccagt cttccgactg
     2401 agcctttcgt tttatttgat gcctggcagt tccctactct cgcgttaacg ctagcatgga
     2461 tgttttccca gtcacgacgt tgtaaaacga cggccagtct taagctcggg cccctacagg
     2521 tcactaatac catctaagta gttgattcat agtgactgga tatgttgtgt tttacagtat
     2581 tatgtagtct gttttttatg caaaatctaa tttaatatat tgatatttat atcattttac
     2641 gtttctcgtt caacttttct atacaaagtt gctatcgatc cgagactagc ctcgagcggc
     2701 cgcccccttc accgagggcc tatttcccat gattccttca tatttgcata tacgatacaa
     2761 ggctgttaga gagataattg gaattaattt gactgtaaac acaaagatat tagtacaaaa
     2821 tacgtgacgt agaaagtaat aatttcttgg gtagtttgca gttttaaaat tatgttttaa
     2881 aatggactat catatgctta ccgtaacttg aaagtatttc gatttcttgg ctttatatat
     2941 cttgtggaaa ggacgaaaca ccggtcctaa ggttaagtcg ccctcgctcg agcgagggcg
     3001 acttaacctt aggtttttg
//
