LOCUS Exported 3318 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION pMVP L5-L4 entry plasmid, contains CMV promoter for 4-component MultiSite Gateway Pro assembly. ACCESSION . VERSION . KEYWORDS IH401: pMVP (L5-L4) CMV promoter SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 3318) 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 3318) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..3318 /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=attL5 /bound_moiety="LR Clonase(TM)" /note="recombination site for the Gateway(R) LR reaction" enhancer 821..1200 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 1201..1404 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 1355..1375 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" primer_bind 1401..1425 /label=LNCX /note="Human CMV promoter, forward primer" protein_bind complement(1433..1528) /gene="mutant version of attL" /label=attL4 /bound_moiety="LR Clonase(TM)" /note="recombination site for the Gateway(R) LR reaction" primer_bind complement(1559..1578) /label=T7 /note="T7 promoter, forward primer" promoter complement(1560..1578) /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" primer_bind complement(1583..1599) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(1583..1599) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(1701..1720) /label=pENTR-R /note="pENTR vectors, reverse primer" CDS 1712..2521 /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(1779..1798) /label=Kan-R /note="Kanamycin resistance gene, reverse primer" rep_origin 2668..3256 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 3157..3176 /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 tgtatacaaa 661 agttgtacga tctatacatt gaatcaatat tggcaattag ccatattagt cattggttat 721 atagcataaa tcaatattgg ctattggcca ttgcatacgt tgtatctata tcataatatg 781 tacatttata ttggctcatg tccaatatga ccgccatgtt gacattgatt attgactagt 841 tattaatagt aatcaattac ggggtcatta gttcatagcc catatatgga gttccgcgtt 901 acataactta cggtaaatgg cccgcctggc tgaccgccca acgacccccg cccattgacg 961 tcaataatga cgtatgttcc catagtaacg ccaataggga ctttccattg acgtcaatgg 1021 gtggagtatt tacggtaaac tgcccacttg gcagtacatc aagtgtatca tatgccaagt 1081 ccgcccccta ttgacgtcaa tgacggtaaa tggcccgcct ggcattatgc ccagtacatg 1141 accttacggg actttcctac ttggcagtac atctacgtat tagtcatcgc tattaccatg 1201 gtgatgcggt tttggcagta caccaatggg cgtggatagc ggtttgactc acggggattt 1261 ccaagtctcc accccattga cgtcaatggg agtttgtttt ggcaccaaaa tcaacgggac 1321 tttccaaaat gtcgtaataa ccccgccccg ttgacgcaaa tgggcggtag gcgtgtacgg 1381 tgggaggtct atataagcag agctcgttta gtgaaccgtc agatccggca cccaactttt 1441 ctatacaaag ttggcattat aagaaagcat tgcttatcaa tttgttgcaa cgaacaggtc 1501 actatcagtc aaaataaaat cattatttgc catccagctg cagggcggcc gcgatatccc 1561 ctatagtgag tcgtattaca tggtcatagc tgtttcctgg cagctctggc ccgtgtctca 1621 aaatctctga tgttacattg cacaagataa aaatatatca tcatgaacaa taaaactgtc 1681 tgcttacata aacagtaata caaggggtgt tatgagccat attcaacggg aaacgtcgag 1741 gccgcgatta aattccaaca tggatgctga tttatatggg tataaatggg ctcgcgataa 1801 tgtcgggcaa tcaggtgcga caatctatcg cttgtatggg aagcccgatg cgccagagtt 1861 gtttctgaaa catggcaaag gtagcgttgc caatgatgtt acagatgaga tggtcagact 1921 aaactggctg acggaattta tgcctcttcc gaccatcaag cattttatcc gtactcctga 1981 tgatgcatgg ttactcacca ctgcgatccc cggaaaaaca gcattccagg tattagaaga 2041 atatcctgat tcaggtgaaa atattgttga tgcgctggca gtgttcctgc gccggttgca 2101 ttcgattcct gtttgtaatt gtccttttaa cagcgatcgc gtatttcgtc tcgctcaggc 2161 gcaatcacga atgaataacg gtttggttga tgcgagtgat tttgatgacg agcgtaatgg 2221 ctggcctgtt gaacaagtct ggaaagaaat gcataaactt ttgccattct caccggattc 2281 agtcgtcact catggtgatt tctcacttga taaccttatt tttgacgagg ggaaattaat 2341 aggttgtatt gatgttggac gagtcggaat cgcagaccga taccaggatc ttgccatcct 2401 atggaactgc ctcggtgagt tttctccttc attacagaaa cggctttttc aaaaatatgg 2461 tattgataat cctgatatga ataaattgca gtttcatttg atgctcgatg agtttttcta 2521 atcagaattg gttaattggt tgtaacactg gcagagcatt acgctgactt gacgggacgg 2581 cgcaagctca tgaccaaaat cccttaacgt gagttacgcg tcgttccact gagcgtcaga 2641 ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg 2701 cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc 2761 aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct 2821 agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc 2881 tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt 2941 ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg 3001 cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct 3061 atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag 3121 ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag 3181 tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg 3241 gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg 3301 gccttttgct cacatgtt //