LOCUS Exported 2893 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION pMVP L1-R5 entry plasmid, contains empty H1-driven shRNA expression cassette for 4-component MultiSite Gateway Pro assembly. ACCESSION . VERSION . KEYWORDS HE401: pMVP (L1-R5) shRNA SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 2893) 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 2893) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..2893 /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 569..668 /gene="mutant version of attL" /label=attL1 /bound_moiety="LR Clonase(TM)" /note="recombination site for the Gateway(R) LR reaction" promoter 689..903 /label=H1 promoter /note="human H1 RNA promoter" primer_bind 824..843 /label=H1 /note="Human H1 promoter, forward primer" protein_bind 956..1079 /gene="mutant version of attR" /label=attR5 /bound_moiety="LR Clonase(TM)" /note="recombination site for the Gateway(R) LR reaction" primer_bind complement(1134..1153) /label=T7 /note="T7 promoter, forward primer" promoter complement(1135..1153) /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" primer_bind complement(1158..1174) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(1158..1174) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(1276..1295) /label=pENTR-R /note="pENTR vectors, reverse primer" CDS 1287..2096 /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(1354..1373) /label=Kan-R /note="Kanamycin resistance gene, reverse primer" rep_origin 2243..2831 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 2732..2751 /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 tgtacaaaaa 661 agcaggctta ggccatggat ccgaattcga acgctgacgt catcaacccg ctccaaggaa 721 tcgcgggccc agtgtcacta ggcgggaaca cccagcgcgc gtgcgccctg gcaggaagat 781 ggctgtgagg gacaggggag tggcgccctg caatatttgc atgtcgctat gtgttctggg 841 aaatcaccat aaacgtgaaa tgtctttgga tttgggaatc ttataagttc tgtatgagac 901 cacagatctg cgtttaaacg gaagctttag gccgcctaga gggaaggtgc tgagacaact 961 ttgtatacaa aagttgaacg agaaacgtaa aatgatataa atatcaatat attaaattag 1021 attttgcata aaaaacagac tacataatac tgtaaaacac aacatatcca gtcactatga 1081 atcaactact tagatggtat tagtgacctg tactgcaggg cggccgcgat atcccctata 1141 gtgagtcgta ttacatggtc atagctgttt cctggcagct ctggcccgtg tctcaaaatc 1201 tctgatgtta cattgcacaa gataaaaata tatcatcatg aacaataaaa ctgtctgctt 1261 acataaacag taatacaagg ggtgttatga gccatattca acgggaaacg tcgaggccgc 1321 gattaaattc caacatggat gctgatttat atgggtataa atgggctcgc gataatgtcg 1381 ggcaatcagg tgcgacaatc tatcgcttgt atgggaagcc cgatgcgcca gagttgtttc 1441 tgaaacatgg caaaggtagc gttgccaatg atgttacaga tgagatggtc agactaaact 1501 ggctgacgga atttatgcct cttccgacca tcaagcattt tatccgtact cctgatgatg 1561 catggttact caccactgcg atccccggaa aaacagcatt ccaggtatta gaagaatatc 1621 ctgattcagg tgaaaatatt gttgatgcgc tggcagtgtt cctgcgccgg ttgcattcga 1681 ttcctgtttg taattgtcct tttaacagcg atcgcgtatt tcgtctcgct caggcgcaat 1741 cacgaatgaa taacggtttg gttgatgcga gtgattttga tgacgagcgt aatggctggc 1801 ctgttgaaca agtctggaaa gaaatgcata aacttttgcc attctcaccg gattcagtcg 1861 tcactcatgg tgatttctca cttgataacc ttatttttga cgaggggaaa ttaataggtt 1921 gtattgatgt tggacgagtc ggaatcgcag accgatacca ggatcttgcc atcctatgga 1981 actgcctcgg tgagttttct ccttcattac agaaacggct ttttcaaaaa tatggtattg 2041 ataatcctga tatgaataaa ttgcagtttc atttgatgct cgatgagttt ttctaatcag 2101 aattggttaa ttggttgtaa cactggcaga gcattacgct gacttgacgg gacggcgcaa 2161 gctcatgacc aaaatccctt aacgtgagtt acgcgtcgtt ccactgagcg tcagaccccg 2221 tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc 2281 aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc 2341 tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt 2401 agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc 2461 taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact 2521 caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac 2581 agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag 2641 aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg 2701 gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg 2761 tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga 2821 gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt 2881 ttgctcacat gtt //