LOCUS Exported 3447 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION pMVP L3-L2 entry plasmid, contains AID + WPRE for 3- or 4-component MultiSite Gateway Pro assembly. Allows fusion of minimal C-term AID domain (activated by auxin) to gene in lentivirus vectors. ACCESSION . VERSION . KEYWORDS LA802: pMVP (L3-L2) AID + WPRE SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 3447) AUTHORS Haldeman JM, Conway AE, Arlotto ME, Slentz DH, Muoio DM, Becker TC, Newgard CB TITLE Creation of versatile cloning platforms for transgene expression and dCas9-based epigenome editing. JOURNAL Nucleic Acids Res. 2018 Dec 27. pii: 5264291. doi: 10.1093/nar/gky1286. PUBMED 30590691 REFERENCE 2 (bases 1 to 3447) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..3447 /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 570..665 /gene="mutant version of attL" /label=attL3 /bound_moiety="LR Clonase(TM)" /note="recombination site for the Gateway(R) LR reaction" CDS 680..883 /codon_start=1 /product="minimal degron from the auxin-responsive IAA17 protein of Arabidopsis thaliana (Kubota et al., 2013)" /label=mini-AID /note="acts as an auxin-inducible degron (AID) in the presence of TIR1" /translation="KEKSACPKDPAKPPAKAQVVGWPPVRSYRKNVMVSCQKSSGGPEA AAFVKVSMDGAPYLRKIDLRMYK" misc_feature 943..1531 /label=WPRE /note="woodchuck hepatitis virus posttranscriptional regulatory element" primer_bind complement(996..1016) /label=WPRE-R /note="WPRE, reverse primer" CDS complement(1414..1425) /codon_start=1 /product="Factor Xa recognition and cleavage site" /label=Factor Xa site /translation="IEGR" protein_bind complement(1559..1658) /gene="mutant version of attL" /label=attL2 /bound_moiety="LR Clonase(TM)" /note="recombination site for the Gateway(R) LR reaction" primer_bind complement(1688..1707) /label=T7 /note="T7 promoter, forward primer" promoter complement(1689..1707) /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" primer_bind complement(1712..1728) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(1712..1728) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(1830..1849) /label=pENTR-R /note="pENTR vectors, reverse primer" CDS 1841..2650 /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(1908..1927) /label=Kan-R /note="Kanamycin resistance gene, reverse primer" rep_origin 2797..3385 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 3286..3305 /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 tcgggcccca aataatgatt ttattttgac tgatagtgac 601 ctgttcgttg caacaaattg atgagcaatg cttttttata atgccaactt tgtataataa 661 agttggtggc ggcggatcca aggagaagag tgcttgtcct aaagatccag ccaaacctcc 721 ggccaaggca caagttgtgg gatggccacc ggtgagatca taccggaaga acgtgatggt 781 ttcctgccaa aaatcaagcg gtggcccgga ggcggcggcg ttcgtgaagg tatcaatgga 841 cggagcaccg tacttgagga aaatcgattt gaggatgtat aaataagaat tctgaagctt 901 agcctagagc tcctatagtg agaccggtta gtaatgatcg acaatcaacc tctggattac 961 aaaatttgtg aaagattgac tggtattctt aactatgttg ctccttttac gctatgtgga 1021 tacgctgctt taatgccttt gtatcatgct attgcttccc gtatggcttt cattttctcc 1081 tccttgtata aatcctggtt gctgtctctt tatgaggagt tgtggcccgt tgtcaggcaa 1141 cgtggcgtgg tgtgcactgt gtttgctgac gcaaccccca ctggttgggg cattgccacc 1201 acctgtcagc tcctttccgg gactttcgct ttccccctcc ctattgccac ggcggaactc 1261 atcgccgcct gccttgcccg ctgctggaca ggggctcggc tgttgggcac tgacaattcc 1321 gtggtgttgt cggggaagct gacgtccttt ccatggctgc tcgcctgtgt tgccacctgg 1381 attctgcgcg ggacgtcctt ctgctacgtc ccttcggccc tcaatccagc ggaccttcct 1441 tcccgcggcc tgctgccggc tctgcggcct cttccgcgtc ttcgccttcg ccctcagacg 1501 agtcggatct ccctttgggc cgcctccccg cctggcgatg gtactgtcct agtacctgac 1561 ccagctttct tgtacaaagt tggcattata agaaagcatt gcttatcaat ttgttgcaac 1621 gaacaggtca ctatcagtca aaataaaatc attatttgcc atccagctgc agggcggccg 1681 cgatatcccc tatagtgagt cgtattacat ggtcatagct gtttcctggc agctctggcc 1741 cgtgtctcaa aatctctgat gttacattgc acaagataaa aatatatcat catgaacaat 1801 aaaactgtct gcttacataa acagtaatac aaggggtgtt atgagccata ttcaacggga 1861 aacgtcgagg ccgcgattaa attccaacat ggatgctgat ttatatgggt ataaatgggc 1921 tcgcgataat gtcgggcaat caggtgcgac aatctatcgc ttgtatggga agcccgatgc 1981 gccagagttg tttctgaaac atggcaaagg tagcgttgcc aatgatgtta cagatgagat 2041 ggtcagacta aactggctga cggaatttat gcctcttccg accatcaagc attttatccg 2101 tactcctgat gatgcatggt tactcaccac tgcgatcccc ggaaaaacag cattccaggt 2161 attagaagaa tatcctgatt caggtgaaaa tattgttgat gcgctggcag tgttcctgcg 2221 ccggttgcat tcgattcctg tttgtaattg tccttttaac agcgatcgcg tatttcgtct 2281 cgctcaggcg caatcacgaa tgaataacgg tttggttgat gcgagtgatt ttgatgacga 2341 gcgtaatggc tggcctgttg aacaagtctg gaaagaaatg cataaacttt tgccattctc 2401 accggattca gtcgtcactc atggtgattt ctcacttgat aaccttattt ttgacgaggg 2461 gaaattaata ggttgtattg atgttggacg agtcggaatc gcagaccgat accaggatct 2521 tgccatccta tggaactgcc tcggtgagtt ttctccttca ttacagaaac ggctttttca 2581 aaaatatggt attgataatc ctgatatgaa taaattgcag tttcatttga tgctcgatga 2641 gtttttctaa tcagaattgg ttaattggtt gtaacactgg cagagcatta cgctgacttg 2701 acgggacggc gcaagctcat gaccaaaatc ccttaacgtg agttacgcgt cgttccactg 2761 agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt 2821 aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca 2881 agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac 2941 tgttcttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac 3001 atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct 3061 taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg 3121 gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga gatacctaca 3181 gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt 3241 aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta 3301 tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc 3361 gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc 3421 cttttgctgg ccttttgctc acatgtt //