LOCUS Exported 3448 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION MuLE (Multiple Lentiviral Expression) Entry vector containing a CMV promoter and multiple cloning site module. Compatible with MultiSite Gateway cloning. ACCESSION . VERSION . KEYWORDS pMuLE ENTR CMV R4-R3 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 3448) 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 3448) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..3448 /organism="synthetic DNA construct" /mol_type="other DNA" primer_bind 92..109 /label=L4440 /note="L4440 vector, forward primer" terminator 268..295 /label=rrnB T2 terminator /note="transcription terminator T2 from the E. coli rrnB gene" terminator 387..473 /gene="Escherichia coli rrnB" /label=rrnB T1 terminator /note="transcription terminator T1 from the E. coli rrnB gene" primer_bind 522..544 /label=M13/pUC Forward /note="In lacZ gene" primer_bind 536..553 /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind 537..553 /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" protein_bind complement(603..727) /gene="mutant version of attR" /label=attR4 /bound_moiety="LR Clonase(TM)" /note="recombination site for the Gateway(R) LR reaction" enhancer 754..1133 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 1134..1337 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 1288..1308 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" promoter 1382..1400 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" protein_bind 1511..1634 /gene="mutant version of attR" /label=attR3 /bound_moiety="LR Clonase(TM)" /note="recombination site for the Gateway(R) LR reaction" promoter complement(1690..1708) /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" primer_bind complement(1713..1729) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(1713..1729) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(1831..1850) /label=pENTR-R /note="pENTR vectors, reverse primer" CDS 1842..2651 /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(1909..1928) /label=Kan-R /note="Kanamycin resistance gene, reverse primer" rep_origin 2798..3386 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 3287..3306 /label=pBR322ori-F /note="pBR322 origin, forward primer" ORIGIN 1 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga 61 taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga 121 gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctggca 181 cgacaggttt cccgactgga aagcgggcag tgagcgcaac gcaattaata cgcgtaccgc 241 tagccaggaa gagtttgtag aaacgcaaaa aggccatccg tcaggatggc cttctgctta 301 gtttgatgcc tggcagttta tggcgggcgt cctgcccgcc accctccggg ccgttgcttc 361 acaacgttca aatccgctcc cggcggattt gtcctactca ggagagcgtt caccgacaaa 421 caacagataa aacgaaaggc ccagtcttcc gactgagcct ttcgttttat ttgatgcctg 481 gcagttccct actctcgcgt taacgctagc atggatgttt tcccagtcac gacgttgtaa 541 aacgacggcc agtcttaagc tcgggcccct acaggtcact aataccatct aagtagttga 601 ttcatagtga ctggatatgt tgtgttttac agtattatgt agtctgtttt ttatgcaaaa 661 tctaatttaa tatattgata tttatatcat tttacgtttc tcgttcaact tttctataca 721 aagttggcga tgtacgggcc agatatacgc gttgacattg attattgact agttattaat 781 agtaatcaat tacggggtca ttagttcata gcccatatat ggagttccgc gttacataac 841 ttacggtaaa tggcccgcct ggctgaccgc ccaacgaccc ccgcccattg acgtcaataa 901 tgacgtatgt tcccatagta acgccaatag ggactttcca ttgacgtcaa tgggtggagt 961 atttacggta aactgcccac ttggcagtac atcaagtgta tcatatgcca agtacgcccc 1021 ctattgacgt caatgacggt aaatggcccg cctggcatta tgcccagtac atgaccttat 1081 gggactttcc tacttggcag tacatctacg tattagtcat cgctattacc atggtgatgc 1141 ggttttggca gtacatcaat gggcgtggat agcggtttga ctcacgggga tttccaagtc 1201 tccaccccat tgacgtcaat gggagtttgt tttggcacca aaatcaacgg gactttccaa 1261 aatgtcgtaa caactccgcc ccattgacgc aaatgggcgg taggcgtgta cggtgggagg 1321 tctatataag cagagctctc tggctaacta gagaacccac tgcttactgg cttatcgaaa 1381 ttaatacgac tcactatagg gagacccaag cttggtaccg agctcggatc cactagtaac 1441 ggccgccagt gtgctggaat tctgcagata tccatcacac tggcggccgc tcgagcatgc 1501 atctagagga caactttgta taataaagtt gaacgagaaa cgtaaaatga tataaatatc 1561 aatatattaa attagatttt gcataaaaaa cagactacat aatactgtaa aacacaacat 1621 atccagtcac tatgaatcaa ctacttagat ggtattagtg acctgtactg cagggcggcc 1681 gcgatatccc ctatagtgag tcgtattaca tggtcatagc tgtttcctgg cagctctggc 1741 ccgtgtctca aaatctctga tgttacattg cacaagataa aaatatatca tcatgaacaa 1801 taaaactgtc tgcttacata aacagtaata caaggggtgt tatgagccat attcaacggg 1861 aaacgtcgag gccgcgatta aattccaaca tggatgctga tttatatggg tataaatggg 1921 ctcgcgataa tgtcgggcaa tcaggtgcga caatctatcg cttgtatggg aagcccgatg 1981 cgccagagtt gtttctgaaa catggcaaag gtagcgttgc caatgatgtt acagatgaga 2041 tggtcagact aaactggctg acggaattta tgcctcttcc gaccatcaag cattttatcc 2101 gtactcctga tgatgcatgg ttactcacca ctgcgatccc cggaaaaaca gcattccagg 2161 tattagaaga atatcctgat tcaggtgaaa atattgttga tgcgctggca gtgttcctgc 2221 gccggttgca ttcgattcct gtttgtaatt gtccttttaa cagcgatcgc gtatttcgtc 2281 tcgctcaggc gcaatcacga atgaataacg gtttggttga tgcgagtgat tttgatgacg 2341 agcgtaatgg ctggcctgtt gaacaagtct ggaaagaaat gcataaactt ttgccattct 2401 caccggattc agtcgtcact catggtgatt tctcacttga taaccttatt tttgacgagg 2461 ggaaattaat aggttgtatt gatgttggac gagtcggaat cgcagaccga taccaggatc 2521 ttgccatcct atggaactgc ctcggtgagt tttctccttc attacagaaa cggctttttc 2581 aaaaatatgg tattgataat cctgatatga ataaattgca gtttcatttg atgctcgatg 2641 agtttttcta atcagaattg gttaattggt tgtaacactg gcagagcatt acgctgactt 2701 gacgggacgg cgcaagctca tgaccaaaat cccttaacgt gagttacgcg tcgttccact 2761 gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg 2821 taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc 2881 aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata 2941 ctgttcttct agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta 3001 catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc 3061 ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg 3121 ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac 3181 agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg 3241 taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt 3301 atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct 3361 cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg 3421 ccttttgctg gccttttgct cacatgtt //