LOCUS Exported 3256 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION pMVP L3-L2 entry plasmid, contains HA epitope tag + WPRE for 3- or 4-component MultiSite Gateway Pro assembly. Allows C-term epitope fusion to a gene of interest in lentivirus vectors.. ACCESSION . VERSION . KEYWORDS KH101: pMVP (L3-L2) HA epitope tag + WPRE SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 3256) 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 3256) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..3256 /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 677..703 /codon_start=1 /product="HA (human influenza hemagglutinin) epitope tag" /label=HA /translation="YPYDVPDYA" misc_feature 752..1340 /label=WPRE /note="woodchuck hepatitis virus posttranscriptional regulatory element" primer_bind complement(805..825) /label=WPRE-R /note="WPRE, reverse primer" CDS complement(1223..1234) /codon_start=1 /product="Factor Xa recognition and cleavage site" /label=Factor Xa site /translation="IEGR" protein_bind complement(1368..1467) /gene="mutant version of attL" /label=attL2 /bound_moiety="LR Clonase(TM)" /note="recombination site for the Gateway(R) LR reaction" primer_bind complement(1497..1516) /label=T7 /note="T7 promoter, forward primer" promoter complement(1498..1516) /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" primer_bind complement(1521..1537) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(1521..1537) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(1639..1658) /label=pENTR-R /note="pENTR vectors, reverse primer" CDS 1650..2459 /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(1717..1736) /label=Kan-R /note="Kanamycin resistance gene, reverse primer" rep_origin 2606..3194 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 3095..3114 /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 ggatcctacc catacgatgt gccagattac gcttaagaat tcctagagct 721 cctatagtga gaccggttag taatgatcga caatcaacct ctggattaca aaatttgtga 781 aagattgact ggtattctta actatgttgc tccttttacg ctatgtggat acgctgcttt 841 aatgcctttg tatcatgcta ttgcttcccg tatggctttc attttctcct ccttgtataa 901 atcctggttg ctgtctcttt atgaggagtt gtggcccgtt gtcaggcaac gtggcgtggt 961 gtgcactgtg tttgctgacg caacccccac tggttggggc attgccacca cctgtcagct 1021 cctttccggg actttcgctt tccccctccc tattgccacg gcggaactca tcgccgcctg 1081 ccttgcccgc tgctggacag gggctcggct gttgggcact gacaattccg tggtgttgtc 1141 ggggaagctg acgtcctttc catggctgct cgcctgtgtt gccacctgga ttctgcgcgg 1201 gacgtccttc tgctacgtcc cttcggccct caatccagcg gaccttcctt cccgcggcct 1261 gctgccggct ctgcggcctc ttccgcgtct tcgccttcgc cctcagacga gtcggatctc 1321 cctttgggcc gcctccccgc ctggcgatgg tactgtccta gtacctgacc cagctttctt 1381 gtacaaagtt ggcattataa gaaagcattg cttatcaatt tgttgcaacg aacaggtcac 1441 tatcagtcaa aataaaatca ttatttgcca tccagctgca gggcggccgc gatatcccct 1501 atagtgagtc gtattacatg gtcatagctg tttcctggca gctctggccc gtgtctcaaa 1561 atctctgatg ttacattgca caagataaaa atatatcatc atgaacaata aaactgtctg 1621 cttacataaa cagtaataca aggggtgtta tgagccatat tcaacgggaa acgtcgaggc 1681 cgcgattaaa ttccaacatg gatgctgatt tatatgggta taaatgggct cgcgataatg 1741 tcgggcaatc aggtgcgaca atctatcgct tgtatgggaa gcccgatgcg ccagagttgt 1801 ttctgaaaca tggcaaaggt agcgttgcca atgatgttac agatgagatg gtcagactaa 1861 actggctgac ggaatttatg cctcttccga ccatcaagca ttttatccgt actcctgatg 1921 atgcatggtt actcaccact gcgatccccg gaaaaacagc attccaggta ttagaagaat 1981 atcctgattc aggtgaaaat attgttgatg cgctggcagt gttcctgcgc cggttgcatt 2041 cgattcctgt ttgtaattgt ccttttaaca gcgatcgcgt atttcgtctc gctcaggcgc 2101 aatcacgaat gaataacggt ttggttgatg cgagtgattt tgatgacgag cgtaatggct 2161 ggcctgttga acaagtctgg aaagaaatgc ataaactttt gccattctca ccggattcag 2221 tcgtcactca tggtgatttc tcacttgata accttatttt tgacgagggg aaattaatag 2281 gttgtattga tgttggacga gtcggaatcg cagaccgata ccaggatctt gccatcctat 2341 ggaactgcct cggtgagttt tctccttcat tacagaaacg gctttttcaa aaatatggta 2401 ttgataatcc tgatatgaat aaattgcagt ttcatttgat gctcgatgag tttttctaat 2461 cagaattggt taattggttg taacactggc agagcattac gctgacttga cgggacggcg 2521 caagctcatg accaaaatcc cttaacgtga gttacgcgtc gttccactga gcgtcagacc 2581 ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct 2641 tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa 2701 ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag 2761 tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc 2821 tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg 2881 actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca 2941 cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat 3001 gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg 3061 tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc 3121 ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc 3181 ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc 3241 cttttgctca catgtt //