LOCUS Exported 3253 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION pMVP L3-L2 entry plasmid, contains FLAG 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 JA001: pMVP (L3-L2) FLAG epitope tag + WPRE SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 3253) 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 3253) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..3253 /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..700 /codon_start=1 /product="FLAG(R) epitope tag, followed by an enterokinase cleavage site" /label=FLAG /translation="DYKDDDDK" misc_feature 749..1337 /label=WPRE /note="woodchuck hepatitis virus posttranscriptional regulatory element" primer_bind complement(802..822) /label=WPRE-R /note="WPRE, reverse primer" CDS complement(1220..1231) /codon_start=1 /product="Factor Xa recognition and cleavage site" /label=Factor Xa site /translation="IEGR" protein_bind complement(1365..1464) /gene="mutant version of attL" /label=attL2 /bound_moiety="LR Clonase(TM)" /note="recombination site for the Gateway(R) LR reaction" primer_bind complement(1494..1513) /label=T7 /note="T7 promoter, forward primer" promoter complement(1495..1513) /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" primer_bind complement(1518..1534) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(1518..1534) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(1636..1655) /label=pENTR-R /note="pENTR vectors, reverse primer" CDS 1647..2456 /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(1714..1733) /label=Kan-R /note="Kanamycin resistance gene, reverse primer" rep_origin 2603..3191 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 3092..3111 /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 ggatccgact acaaggatga cgacgataag taagaattcc tagagctcct 721 atagtgagac cggttagtaa tgatcgacaa tcaacctctg gattacaaaa tttgtgaaag 781 attgactggt attcttaact atgttgctcc ttttacgcta tgtggatacg ctgctttaat 841 gcctttgtat catgctattg cttcccgtat ggctttcatt ttctcctcct tgtataaatc 901 ctggttgctg tctctttatg aggagttgtg gcccgttgtc aggcaacgtg gcgtggtgtg 961 cactgtgttt gctgacgcaa cccccactgg ttggggcatt gccaccacct gtcagctcct 1021 ttccgggact ttcgctttcc ccctccctat tgccacggcg gaactcatcg ccgcctgcct 1081 tgcccgctgc tggacagggg ctcggctgtt gggcactgac aattccgtgg tgttgtcggg 1141 gaagctgacg tcctttccat ggctgctcgc ctgtgttgcc acctggattc tgcgcgggac 1201 gtccttctgc tacgtccctt cggccctcaa tccagcggac cttccttccc gcggcctgct 1261 gccggctctg cggcctcttc cgcgtcttcg ccttcgccct cagacgagtc ggatctccct 1321 ttgggccgcc tccccgcctg gcgatggtac tgtcctagta cctgacccag ctttcttgta 1381 caaagttggc attataagaa agcattgctt atcaatttgt tgcaacgaac aggtcactat 1441 cagtcaaaat aaaatcatta tttgccatcc agctgcaggg cggccgcgat atcccctata 1501 gtgagtcgta ttacatggtc atagctgttt cctggcagct ctggcccgtg tctcaaaatc 1561 tctgatgtta cattgcacaa gataaaaata tatcatcatg aacaataaaa ctgtctgctt 1621 acataaacag taatacaagg ggtgttatga gccatattca acgggaaacg tcgaggccgc 1681 gattaaattc caacatggat gctgatttat atgggtataa atgggctcgc gataatgtcg 1741 ggcaatcagg tgcgacaatc tatcgcttgt atgggaagcc cgatgcgcca gagttgtttc 1801 tgaaacatgg caaaggtagc gttgccaatg atgttacaga tgagatggtc agactaaact 1861 ggctgacgga atttatgcct cttccgacca tcaagcattt tatccgtact cctgatgatg 1921 catggttact caccactgcg atccccggaa aaacagcatt ccaggtatta gaagaatatc 1981 ctgattcagg tgaaaatatt gttgatgcgc tggcagtgtt cctgcgccgg ttgcattcga 2041 ttcctgtttg taattgtcct tttaacagcg atcgcgtatt tcgtctcgct caggcgcaat 2101 cacgaatgaa taacggtttg gttgatgcga gtgattttga tgacgagcgt aatggctggc 2161 ctgttgaaca agtctggaaa gaaatgcata aacttttgcc attctcaccg gattcagtcg 2221 tcactcatgg tgatttctca cttgataacc ttatttttga cgaggggaaa ttaataggtt 2281 gtattgatgt tggacgagtc ggaatcgcag accgatacca ggatcttgcc atcctatgga 2341 actgcctcgg tgagttttct ccttcattac agaaacggct ttttcaaaaa tatggtattg 2401 ataatcctga tatgaataaa ttgcagtttc atttgatgct cgatgagttt ttctaatcag 2461 aattggttaa ttggttgtaa cactggcaga gcattacgct gacttgacgg gacggcgcaa 2521 gctcatgacc aaaatccctt aacgtgagtt acgcgtcgtt ccactgagcg tcagaccccg 2581 tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc 2641 aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc 2701 tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt 2761 agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc 2821 taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact 2881 caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac 2941 agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag 3001 aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg 3061 gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg 3121 tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga 3181 gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt 3241 ttgctcacat gtt //