pETM-60-NusA-TEV-HsRCK_296-472_E

Plasmid Information

Purpose

Bacterial Expression of HsRck_296-472. Contains a TEV site rather than a HRV 3C site downstream of NusA.

Depositor

Elisa Izaurralde

Article

Tritschler et al Mol Cell. 2009 Mar 13. 33(5):661-8.

Insert

HsRck_296-472 (DDX6 Human)

Available Since

March 16, 2023

Availability

Available

Sequence Information

Length

7,363 bp

Topology

circular

Molecule Type

other DNA

View on AddGene.org

Features

CDS 7

Label	Gene	Product	Position	Strand
KanR	aph(3')-Ia	aminoglycoside phosphotransferase	558..1374	Rev
DNA Sequence ATGAGCCATATTCAACGGGAAACGTCTTGCTCTAGGCCGCGATTAAATTCCAACATGGATGCTGATTTATATGGGTATAAATGGGCTCGCGATAATGTCGGGCAATCAGGTGCGACAATCTATCGATTGTATGGGAAGCCCGATGCGCCAGAGTTGTTTCTGAAACATGGCAAAGGTAGCGTTGCCAATGATGTTACAGATGAGATGGTCAGACTAAACTGGCTGACGGAATTTATGCCTCTTCCGACCATCAAGCATTTTATCCGTACTCCTGATGATGCATGGTTACTCACCACTGCGATCCCCGGGAAAACAGCATTCCAGGTATTAGAAGAATATCCTGATTCAGGTGAAAATATTGTTGATGCGCTGGCAGTGTTCCTGCGCCGGTTGCATTCGATTCCTGTTTGTAATTGTCCTTTTAACAGCGATCGCGTATTTCGTCTCGCTCAGGCGCAATCACGAATGAATAACGGTTTGGTTGATGCGAGTGATTTTGATGACGAGCGTAATGGCTGGCCTGTTGAACAAGTCTGGAAAGAAATGCATAAACTTTTGCCATTCTCACCGGATTCAGTCGTCACTCATGGTGATTTCTCACTTGATAACCTTATTTTTGACGAGGGGAAATTAATAGGTTGTATTGATGTTGGACGAGTCGGAATCGCAGACCGATACCAGGATCTTGCCATCCTATGGAACTGCCTCGGTGAGTTTTCTCCTTCATTACAGAAACGGCTTTTTCAAAAATATGGTATTGATAATCCTGATATGAATAAATTGCAGTTTCATTTGATGCTCGATGAGTTTTTCTAA Protein Sequence MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF
rop	rop	Rop protein, which maintains plasmids a…	2510..2702	Rev
DNA Sequence GTGACCAAACAGGAAAAAACCGCCCTTAACATGGCCCGCTTTATCAGAAGCCAGACATTAACGCTTCTGGAGAAACTCAACGAGCTGGACGCGGATGAACAGGCAGACATCTGTGAATCGCTTCACGACCACGCTGATGAGCTTTACCGCAGCTGCCTCGCGCGTTTCGGTGATGACGGTGAAAACCTCTGA Protein Sequence MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHADELYRSCLARFGDDGENL
lacI	lacI	lac repressor	3510..4593	Rev
DNA Sequence GTGAAACCAGTAACGTTATACGATGTCGCAGAGTATGCCGGTGTCTCTTATCAGACCGTTTCCCGCGTGGTGAACCAGGCCAGCCACGTTTCTGCGAAAACGCGGGAAAAAGTGGAAGCGGCGATGGCGGAGCTGAATTACATTCCCAACCGCGTGGCACAACAACTGGCGGGCAAACAGTCGTTGCTGATTGGCGTTGCCACCTCCAGTCTGGCCCTGCACGCGCCGTCGCAAATTGTCGCGGCGATTAAATCTCGCGCCGATCAACTGGGTGCCAGCGTGGTGGTGTCGATGGTAGAACGAAGCGGCGTCGAAGCCTGTAAAGCGGCGGTGCACAATCTTCTCGCGCAACGCGTCAGTGGGCTGATCATTAACTATCCGCTGGATGACCAGGATGCCATTGCTGTGGAAGCTGCCTGCACTAATGTTCCGGCGTTATTTCTTGATGTCTCTGACCAGACACCCATCAACAGTATTATTTTCTCCCATGAAGACGGTACGCGACTGGGCGTGGAGCATCTGGTCGCATTGGGTCACCAGCAAATCGCGCTGTTAGCGGGCCCATTAAGTTCTGTCTCGGCGCGTCTGCGTCTGGCTGGCTGGCATAAATATCTCACTCGCAATCAAATTCAGCCGATAGCGGAACGGGAAGGCGACTGGAGTGCCATGTCCGGTTTTCAACAAACCATGCAAATGCTGAATGAGGGCATCGTTCCCACTGCGATGCTGGTTGCCAACGATCAGATGGCGCTGGGCGCAATGCGCGCCATTACCGAGTCCGGGCTGCGCGTTGGTGCGGATATCTCGGTAGTGGGATACGACGATACCGAAGACAGCTCATGTTATATCCCGCCGTTAACCACCATCAAACAGGATTTTCGCCTGCTGGGGCAAACCAGCGTGGACCGCTTGCTGCAACTCTCTCAGGGCCAGGCGGTGAAGGGCAATCAGCTGTTGCCCGTCTCACTGGTGAAAAGAAAAACCACCCTGGCGCCCAATACGCAAACCGCCTCTCCCCGCGCGTTGGCCGATTCATTAATGCAGCTGGCACGACAGGTTTCCCGACTGGAAAGCGGGCAGTGA Protein Sequence MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAELNYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGVEACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSHEDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSAMSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSCYIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPRALADSLMQLARQVSRLESGQ
NusA	nusA	transcription elongation protein (N uti…	5066..6548	Fwd
DNA Sequence ATGAAAGAAATTTTGGCTGTAGTTGAAGCCGTATCCAATGAAAAGGCGCTACCTCGCGAGAAGATTTTCGAAGCATTGGAAAGCGCGCTGGCGACAGCAACAAAGAAAAAATATGAACAAGAGATCGACGTCCGCGTACAGATCGATCGCAAAAGCGGTGATTTTGACACTTTCCGTCGCTGGTTAGTTGTTGATGAAGTCACCCAGCCGACCAAGGAAATCACCCTTGAAGCCGCACGTTATGAAGATGAAAGCCTGAACCTGGGCGATTACGTTGAAGATCAGATTGAGTCTGTTACCTTTGACCGTATCACTACCCAGACGGCAAAACAGGTTATCGTGCAGAAAGTGCGTGAAGCCGAACGTGCGATGGTGGTTGATCAGTTCCGTGAACACGAAGGTGAAATCATCACCGGCGTGGTGAAAAAAGTAAACCGCGACAACATCTCTCTGGATCTGGGCAACAACGCTGAAGCCGTGATCCTGCGCGAAGATATGCTGCCGCGTGAAAACTTCCGCCCTGGCGACCGCGTTCGTGGCGTGCTCTATTCCGTTCGCCCGGAAGCGCGTGGCGCGCAACTGTTCGTCACTCGTTCCAAGCCGGAAATGCTGATCGAACTGTTCCGTATTGAAGTGCCAGAAATCGGCGAAGAAGTGATTGAAATTAAAGCAGCGGCTCGCGATCCGGGTTCTCGTGCGAAAATCGCGGTGAAAACCAACGATAAACGTATCGATCCGGTAGGTGCTTGCGTAGGTATGCGTGGCGCGCGTGTTCAGGCGGTGTCTACTGAACTGGGTGGCGAGCGTATCGATATCGTCCTGTGGGATGATAACCCGGCGCAGTTCGTGATTAACGCAATGGCACCGGCAGACGTTGCTTCTATCGTGGTGGATGAAGATAAACACACTATGGACATCGCCGTTGAAGCCGGTAATCTGGCGCAGGCGATTGGCCGTAACGGTCAGAACGTGCGTCTGGCTTCGCAACTGAGCGGTTGGGAACTCAACGTGATGACCGTTGACGACCTGCAAGCTAAGCATCAGGCGGAAGCGCACGCAGCGATCGACACCTTCACCAAATATCTCGACATCGACGAAGACTTCGCGACTGTTCTGGTAGAAGAAGGCTTCTCGACGCTGGAAGAATTGGCCTATGTGCCGATGAAAGAGCTGTTGGAAATCGAAGGCCTTGATGAGCCGACCGTTGAAGCACTGCGCGAGCGTGCTAAAAATGCACTGGCCACCATTGCACAGGCCCAGGAAGAAAGCCTCGGTGATAACAAACCGGCTGACGATCTGCTGAACCTTGAAGGGGTAGATCGTGATTTGGCATTCAAACTGGCCGCCCGTGGCGTTTGTACGCTGGAAGATCTCGCCGAACAGGGCATTGATGATCTGGCTGATATCGAAGGGTTGACCGACGAAAAAGCCGGAGCACTGATTATGGCTGCCCGTAATATTTGCTGGTTCGGTGACGAAGCG Protein Sequence MKEILAVVEAVSNEKALPREKIFEALESALATATKKKYEQEIDVRVQIDRKSGDFDTFRRWLVVDEVTQPTKEITLEAARYEDESLNLGDYVEDQIESVTFDRITTQTAKQVIVQKVREAERAMVVDQFREHEGEIITGVVKKVNRDNISLDLGNNAEAVILREDMLPRENFRPGDRVRGVLYSVRPEARGAQLFVTRSKPEMLIELFRIEVPEIGEEVIEIKAAARDPGSRAKIAVKTNDKRIDPVGACVGMRGARVQAVSTELGGERIDIVLWDDNPAQFVINAMAPADVASIVVDEDKHTMDIAVEAGNLAQAIGRNGQNVRLASQLSGWELNVMTVDDLQAKHQAEAHAAIDTFTKYLDIDEDFATVLVEEGFSTLEELAYVPMKELLEIEGLDEPTVEALRERAKNALATIAQAQEESLGDNKPADDLLNLEGVDRDLAFKLAARGVCTLEDLAEQGIDDLADIEGLTDEKAGALIMAARNICWFGDEA
6xHis	—	6xHis affinity tag	6563..6581	Fwd
DNA Sequence CATCACCATCACCATCAC Protein Sequence HHHHHH
TEV site	—	tobacco etch virus (TEV) protease recog…	6590..6611	Fwd
DNA Sequence GAGAATCTTTATTTTCAGGGC Protein Sequence ENLYFQG
6xHis	—	6xHis affinity tag	7206..7224	Fwd
DNA Sequence CACCACCACCACCACCAC Protein Sequence HHHHHH

promoter 2

Label	Gene	Product	Position	Strand
lacI promoter	lacI	—	4593..4671	Rev
DNA Sequence GACACCATCGAATGGCGCAAAACCTTTCGCGGTATGGCATGATAGCGCCCGGAAGAGAGTCAATTCAGGGTGGTGAAT
T7 promoter	—	—	4979..4998	Fwd
DNA Sequence TAATACGACTCACTATAGG

protein_bind 1

Label	Gene	Product	Position	Strand
lac operator	—	—	4998..5023	Fwd
DNA Sequence GGAATTGTGAGCGGATAACAATTCC

rep_origin 2

Label	Gene	Product	Position	Strand
f1 ori	—	—	11..467	Fwd
DNA Sequence ACGCGCCCTGTAGCGGCGCATTAAGCGCGGCGGGTGTGGTGGTTACGCGCAGCGTGACCGCTACACTTGCCAGCGCCCTAGCGCCCGCTCCTTTCGCTTTCTTCCCTTCCTTTCTCGCCACGTTCGCCGGCTTTCCCCGTCAAGCTCTAAATCGGGGGCTCCCTTTAGGGTTCCGATTTAGTGCTTTACGGCACCTCGACCCCAAAAAACTTGATTAGGGTGATGGTTCACGTAGTGGGCCATCGCCCTGATAGACGGTTTTTCGCCCTTTGACGTTGGAGTCCACGTTCTTTAATAGTGGACTCTTGTTCCAAACTGGAACAACACTCAACCCTATCTCGGTCTATTCTTTTGATTTATAAGGGATTTTGCCGATTTCGGCCTATTGGTTAAAAAATGAGCTGATTTAACAAAAATTTAACGCGAATTTTAACAAAATATTAACGCTTACAATTT
ori	—	—	1495..2084	Fwd
DNA Sequence TTGAGATCCTTTTTTTCTGCGCGTAATCTGCTGCTTGCAAACAAAAAAACCACCGCTACCAGCGGTGGTTTGTTTGCCGGATCAAGAGCTACCAACTCTTTTTCCGAAGGTAACTGGCTTCAGCAGAGCGCAGATACCAAATACTGTCCTTCTAGTGTAGCCGTAGTTAGGCCACCACTTCAAGAACTCTGTAGCACCGCCTACATACCTCGCTCTGCTAATCCTGTTACCAGTGGCTGCTGCCAGTGGCGATAAGTCGTGTCTTACCGGGTTGGACTCAAGACGATAGTTACCGGATAAGGCGCAGCGGTCGGGCTGAACGGGGGGTTCGTGCACACAGCCCAGCTTGGAGCGAACGACCTACACCGAACTGAGATACCTACAGCGTGAGCTATGAGAAAGCGCCACGCTTCCCGAAGGGAGAAAGGCGGACAGGTATCCGGTAAGCGGCAGGGTCGGAACAGGAGAGCGCACGAGGGAGCTTCCAGGGGGAAACGCCTGGTATCTTTATAGTCCTGTCGGGTTTCGCCACCTCTGACTTGAGCGTCGATTTTTGTGATGCTCGTCAGGGGGGCGGAGCCTATGGAAA

terminator 1

Label	Gene	Product	Position	Strand
T7 terminator	—	—	7290..7338	Fwd
DNA Sequence CTAGCATAACCCCTTGGGGCCTCTAAACGGGTCTTGAGGGGTTTTTTG