LOCUS Exported 3209 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION MuLE (Multiple Lentiviral Expression) Entry vector containing a 7SK RNA polymerase III promoter and miR30-based hairpin module for shRNA expression. Compatible with MultiSite Gateway cloning. ACCESSION . VERSION . KEYWORDS pMuLE ENTR 7SK-miR-30 R4-R3 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 3209) 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 3209) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..3209 /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" ncRNA 2986..3079 /label=5' miR-30a /note="sequence upstream of the 71-nt precursor of the human miR-30a microRNA (Zeng et al., 2002)" /ncRNA_class="miRNA" ncRNA 3093..3205 /label=3' miR-30a /note="sequence downstream of the 71-nt precursor of the human miR-30a microRNA (Zeng et al., 2002)" /ncRNA_class="miRNA" 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 tagtgctgca gtatttagca 2701 tgccccaccc atctgcaagg cattctggat agtgtcaaaa cagccggaaa tcaagtccgt 2761 ttatctcaaa ctttagcatt ttgggaataa atgatatttg ctatgctggt taaattagat 2821 tttagttaaa tttcctgctg aagctctagt acgataagta acttgaccta agtgtaaagt 2881 tgagatttcc ttcaggttta tatagcttgt gcgccgcctg ggtacctcgg tcttcataga 2941 tctgcagcac atatactagt cgactaggga taacagggta attgtttgaa tgaggcttca 3001 gtactttaca gaatcgttgc ctgcacatct tggaaacact tgctgggatt acttcttcag 3061 gttaacccaa cagaaggctc gagcaaccag aattcagggg gctactttag gagcaattat 3121 cttgtttact aaaactgaat accttgctat ctctttgata catttttaca aagctgaatt 3181 aaaatggtat aaattaaatc acttttttc //