LOCUS Exported 3960 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION pMVP L3-L2 entry plasmid, contains eGFP + WPRE for 3- or 4-component MultiSite Gateway Pro assembly. Allows C-term eGFP fusion to gene of interest in lentivirus vectors.. ACCESSION . VERSION . KEYWORDS LC801: pMVP (L3-L2) eGFP + WPRE SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 3960) 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 3960) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..3960 /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..1399 /codon_start=1 /product="the original enhanced GFP (Yang et al., 1996)" /label=EGFP /note="mammalian codon-optimized" /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL EFVTAAGITLGMDELYK" CDS 680..1396 /codon_start=1 /product="enhanced GFP" /label=EGFP /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL EFVTAAGITLGMDELYK" primer_bind complement(725..746) /label=EGFP-N /note="EGFP, reverse primer" primer_bind complement(986..1005) /label=EXFP-R /note="For distinguishing EGFP variants, reverse primer" primer_bind 1333..1354 /label=EGFP-C /note="EGFP, forward primer" misc_feature 1456..2044 /label=WPRE /note="woodchuck hepatitis virus posttranscriptional regulatory element" primer_bind complement(1509..1529) /label=WPRE-R /note="WPRE, reverse primer" CDS complement(1927..1938) /codon_start=1 /product="Factor Xa recognition and cleavage site" /label=Factor Xa site /translation="IEGR" protein_bind complement(2072..2171) /gene="mutant version of attL" /label=attL2 /bound_moiety="LR Clonase(TM)" /note="recombination site for the Gateway(R) LR reaction" primer_bind complement(2201..2220) /label=T7 /note="T7 promoter, forward primer" promoter complement(2202..2220) /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" primer_bind complement(2225..2241) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(2225..2241) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(2343..2362) /label=pENTR-R /note="pENTR vectors, reverse primer" CDS 2354..3163 /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(2421..2440) /label=Kan-R /note="Kanamycin resistance gene, reverse primer" rep_origin 3310..3898 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 3799..3818 /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 tggtgagcaa gggcgaggag ctgttcaccg gggtggtgcc 721 catcctggtc gagctggacg gcgacgtaaa cggccacaag ttcagcgtgt ccggcgaggg 781 cgagggcgat gccacctacg gcaagctgac cctgaagttc atctgcacca ccggcaagct 841 gcccgtgccc tggcccaccc tcgtgaccac cctgacctac ggcgtgcagt gcttcagccg 901 ctaccccgac cacatgaagc agcacgactt cttcaagtcc gccatgcccg aaggctacgt 961 ccaggagcgc accatcttct tcaaggacga cggcaactac aagacccgcg ccgaggtgaa 1021 gttcgagggc gacaccctgg tgaaccgcat cgagctgaag ggcatcgact tcaaggagga 1081 cggcaacatc ctggggcaca agctggagta caactacaac agccacaacg tctatatcat 1141 ggccgacaag cagaagaacg gcatcaaggt gaacttcaag atccgccaca acatcgagga 1201 cggcagcgtg cagctcgccg accactacca gcagaacacc cccatcggcg acggccccgt 1261 gctgctgccc gacaaccact acctgagcac ccagtccgcc ctgagcaaag accccaacga 1321 gaagcgcgat cacatggtcc tgctggagtt cgtgaccgcc gccgggatca ctctcggcat 1381 ggacgagctg tacaagtaag aattctgaag cttagcctag agctcctata gtgagaccgg 1441 ttagtaatga tcgacaatca acctctggat tacaaaattt gtgaaagatt gactggtatt 1501 cttaactatg ttgctccttt tacgctatgt ggatacgctg ctttaatgcc tttgtatcat 1561 gctattgctt cccgtatggc tttcattttc tcctccttgt ataaatcctg gttgctgtct 1621 ctttatgagg agttgtggcc cgttgtcagg caacgtggcg tggtgtgcac tgtgtttgct 1681 gacgcaaccc ccactggttg gggcattgcc accacctgtc agctcctttc cgggactttc 1741 gctttccccc tccctattgc cacggcggaa ctcatcgccg cctgccttgc ccgctgctgg 1801 acaggggctc ggctgttggg cactgacaat tccgtggtgt tgtcggggaa gctgacgtcc 1861 tttccatggc tgctcgcctg tgttgccacc tggattctgc gcgggacgtc cttctgctac 1921 gtcccttcgg ccctcaatcc agcggacctt ccttcccgcg gcctgctgcc ggctctgcgg 1981 cctcttccgc gtcttcgcct tcgccctcag acgagtcgga tctccctttg ggccgcctcc 2041 ccgcctggcg atggtactgt cctagtacct gacccagctt tcttgtacaa agttggcatt 2101 ataagaaagc attgcttatc aatttgttgc aacgaacagg tcactatcag tcaaaataaa 2161 atcattattt gccatccagc tgcagggcgg ccgcgatatc ccctatagtg agtcgtatta 2221 catggtcata gctgtttcct ggcagctctg gcccgtgtct caaaatctct gatgttacat 2281 tgcacaagat aaaaatatat catcatgaac aataaaactg tctgcttaca taaacagtaa 2341 tacaaggggt gttatgagcc atattcaacg ggaaacgtcg aggccgcgat taaattccaa 2401 catggatgct gatttatatg ggtataaatg ggctcgcgat aatgtcgggc aatcaggtgc 2461 gacaatctat cgcttgtatg ggaagcccga tgcgccagag ttgtttctga aacatggcaa 2521 aggtagcgtt gccaatgatg ttacagatga gatggtcaga ctaaactggc tgacggaatt 2581 tatgcctctt ccgaccatca agcattttat ccgtactcct gatgatgcat ggttactcac 2641 cactgcgatc cccggaaaaa cagcattcca ggtattagaa gaatatcctg attcaggtga 2701 aaatattgtt gatgcgctgg cagtgttcct gcgccggttg cattcgattc ctgtttgtaa 2761 ttgtcctttt aacagcgatc gcgtatttcg tctcgctcag gcgcaatcac gaatgaataa 2821 cggtttggtt gatgcgagtg attttgatga cgagcgtaat ggctggcctg ttgaacaagt 2881 ctggaaagaa atgcataaac ttttgccatt ctcaccggat tcagtcgtca ctcatggtga 2941 tttctcactt gataacctta tttttgacga ggggaaatta ataggttgta ttgatgttgg 3001 acgagtcgga atcgcagacc gataccagga tcttgccatc ctatggaact gcctcggtga 3061 gttttctcct tcattacaga aacggctttt tcaaaaatat ggtattgata atcctgatat 3121 gaataaattg cagtttcatt tgatgctcga tgagtttttc taatcagaat tggttaattg 3181 gttgtaacac tggcagagca ttacgctgac ttgacgggac ggcgcaagct catgaccaaa 3241 atcccttaac gtgagttacg cgtcgttcca ctgagcgtca gaccccgtag aaaagatcaa 3301 aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc 3361 accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt 3421 aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc cgtagttagg 3481 ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc 3541 agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt 3601 accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga 3661 gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct 3721 tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg 3781 cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca 3841 cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa 3901 cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt //