MT4_C30Y R31W — PlasmidDB

Plasmid Information

Purpose

Bacterial expression plasmids for production of recombinant metallothionein 3 subisoform with C30Y and R31W substitution

Depositor

Artur Krężel

Insert

Metallothionein 4 (MT4 Human)

Sequence Information

Length

7,099 bp

Topology

circular

Molecule Type

other DNA

View on AddGene.org

Features

CDS 6

Label	Gene	Product	Position	Strand
AmpR	bla	beta-lactamase	118..979	Fwd
DNA Sequence ATGAGTATTCAACATTTCCGTGTCGCCCTTATTCCCTTTTTTGCGGCATTTTGCCTTCCTGTTTTTGCTCACCCAGAAACGCTGGTGAAAGTAAAAGATGCTGAAGATCAGTTGGGTGCACGAGTGGGTTACATCGAACTGGATCTCAACAGCGGTAAGATCCTTGAGAGTTTTCGCCCCGAAGAACGTTTCCCAATGATGAGCACTTTTAAAGTTCTGCTATGTGGCGCGGTATTATCCCGTGTTGACGCCGGGCAAGAGCAACTCGGTCGCCGCATACACTATTCTCAGAATGACTTGGTTGAGTACTCACCAGTCACAGAAAAGCATCTTACGGATGGCATGACAGTAAGAGAATTATGCAGTGCTGCCATAACCATGAGTGATAACACTGCGGCCAACTTACTTCTGACAACGATCGGAGGACCGAAGGAGCTAACCGCTTTTTTGCACAACATGGGGGATCATGTAACTCGCCTTGATCGTTGGGAACCGGAGCTGAATGAAGCCATACCAAACGACGAGCGTGACACCACGATGCCTGTAGCAATGGCAACAACGTTGCGCAAACTATTAACTGGCGAACTACTTACTCTAGCTTCCCGGCAACAATTAATAGACTGGATGGAGGCGGATAAAGTTGCAGGACCACTTCTGCGCTCGGCCCTTCCGGCTGGCTGGTTTATTGCTGATAAATCTGGAGCCGGTGAGCGTGGGTCTCGCGGTATCATTGCAGCACTGGGGCCAGATGGTAAGCCCTCCCGTATCGTAGTTATCTACACGACGGGGAGTCAGGCAACTATGGATGAACGAAATAGACAGATCGCTGAGATAGGTGCCTCACTGATTAAGCATTGGTAA Protein Sequence MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYIELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYSPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRWEPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGASLIKHW
rop	rop	Rop protein, which maintains plasmids a…	2601..2793	Rev
DNA Sequence GTGACCAAACAGGAAAAAACCGCCCTTAACATGGCCCGCTTTATCAGAAGCCAGACATTAACGCTTCTGGAGAAACTCAACGAGCTGGACGCGGATGAACAGGCAGACATCTGTGAATCGCTGCACGACCACGCTGATGAGCTTTACCGCAGCTGCCTCGCGCGTTTCGGTGATGACGGTGAAAACCTCTGA Protein Sequence MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHADELYRSCLARFGDDGENL
lacI	lacI	lac repressor	3349..4432	Rev
DNA Sequence GTGAAACCAGTAACGTTATACGATGTCGCAGAGTATGCCGGTGTCTCTTATCAGACCGTTTCCCGCGTGGTGAACCAGGCCAGCCACGTTTCTGCGAAAACGCGGGAAAAAGTGGAAGCGGCGATGGCGGAGCTGAATTACATTCCCAACCGCGTGGCACAACAACTGGCGGGCAAACAGTCGTTGCTGATTGGCGTTGCCACCTCCAGTCTGGCCCTGCACGCGCCGTCGCAAATTGTCGCGGCGATTAAATCTCGCGCCGATCAACTGGGTGCCAGCGTGGTGGTGTCGATGGTAGAACGAAGCGGCGTCGAAGCCTGTAAAGCGGCGGTGCACAATCTTCTCGCGCAACGCGTCAGTGGGCTGATCATTAACTATCCGCTGGATGACCAGGATGCCATTGCTGTGGAAGCTGCCTGCACTAATGTTCCGGCGTTATTTCTTGATGTCTCTGACCAGACACCCATCAACAGTATTATTTTCTCCCATGAAGACGGTACGCGACTGGGCGTGGAGCATCTGGTCGCATTGGGTCACCAGCAAATCGCGCTGTTAGCGGGCCCATTAAGTTCTGTCTCGGCGCGTCTGCGTCTGGCTGGCTGGCATAAATATCTCACTCGCAATCAAATTCAGCCGATAGCGGAACGGGAAGGCGACTGGAGTGCCATGTCCGGTTTTCAACAAACCATGCAAATGCTGAATGAGGGCATCGTTCCCACTGCGATGCTGGTTGCCAACGATCAGATGGCGCTGGGCGCAATGCGCGCCATTACCGAGTCCGGGCTGCGCGTTGGTGCGGACATCTCGGTAGTGGGATACGACGATACCGAAGACAGCTCATGTTATATCCCGCCGTTAACCACCATCAAACAGGATTTTCGCCTGCTGGGGCAAACCAGCGTGGACCGCTTGCTGCAACTCTCTCAGGGCCAGGCGGTGAAGGGCAATCAGCTGTTGCCCGTCTCACTGGTGAAAAGAAAAACCACCCTGGCGCCCAATACGCAAACCGCCTCTCCCCGCGCGTTGGCCGATTCATTAATGCAGCTGGCACGACAGGTTTCCCGACTGGAAAGCGGGCAGTGA Protein Sequence MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAELNYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGVEACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSHEDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSAMSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSCYIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPRALADSLMQLARQVSRLESGQ
Sce VMA intein 5' region	Saccharomyces cerevisiae VMA1	5' region of the intein from the yeast …	5199..6018	Fwd
DNA Sequence TGCTTTGCCAAGGGTACCAATGTTTTAATGGCGGATGGGTCTATTGAATGTATTGAAAACATTGAGGTTGGTAATAAGGTCATGGGTAAAGATGGCAGACCTCGTGAGGTAATTAAATTGCCCAGAGGAAGAGAAACTATGTACAGCGTCGTGCAGAAAAGTCAGCACAGAGCCCACAAAAGTGACTCAAGTCGTGAAGTGCCAGAATTACTCAAGTTTACGTGTAATGCGACCCATGAGTTGGTTGTTAGAACACCTCGTAGTGTCCGCCGTTTGTCTCGTACCATTAAGGGTGTCGAATATTTTGAAGTTATTACTTTTGAGATGGGCCAAAAGAAAGCCCCCGACGGTAGAATTGTTGAGCTTGTCAAGGAAGTTTCAAAGAGCTACCCAATATCTGAGGGGCCTGAGAGAGCCAACGAATTAGTAGAATCCTATAGAAAGGCTTCAAATAAAGCTTATTTTGAGTGGACTATTGAGGCCAGAGATCTTTCTCTGTTGGGTTCCCATGTTCGTAAAGCTACCTACCAGACTTACGCTCCAATTCTTTATGAGAATGACCACTTTTTCGACTACATGCAAAAAAGTAAGTTTCATCTCACCATTGAAGGTCCAAAAGTACTTGCTTATTTACTTGGTTTATGGATTGGTGATGGATTGTCTGACAGGGCAACTTTTTCGGTTGATTCCAGAGATACTTCTTTGATGGAACGTGTTACTGAATATGCTGAAAAGTTGAATTTGTGCGCCGAGTATAAGGACAGAAAAGAACCACAAGTTGCCAAAACTGTTAATTTGTACTCTAAAGTTGTCAGAGGT Protein Sequence CFAKGTNVLMADGSIECIENIEVGNKVMGKDGRPREVIKLPRGRETMYSVVQKSQHRAHKSDSSREVPELLKFTCNATHELVVRTPRSVRRLSRTIKGVEYFEVITFEMGQKKAPDGRIVELVKEVSKSYPISEGPERANELVESYRKASNKAYFEWTIEARDLSLLGSHVRKATYQTYAPILYENDHFFDYMQKSKFHLTIEGPKVLAYLLGLWIGDGLSDRATFSVDSRDTSLMERVTEYAEKLNLCAEYKDRKEPQVAKTVNLYSKVVRG
CBD	Bacillus circulans chiA	chitin binding domain from chitinase A1…	6024..6177	Fwd
DNA Sequence ACAAATCCTGGTGTATCCGCTTGGCAGGTCAACACAGCTTATACTGCGGGACAATTGGTCACATATAACGGCAAGACGTATAAATGTTTGCAGCCCCACACCTCCTTGGCAGGATGGGAACCATCCAACGTTCCTGCCTTGTGGCAGCTTCAA Protein Sequence TNPGVSAWQVNTAYTAGQLVTYNGKTYKCLQPHTSLAGWEPSNVPALWQLQ
Sce VMA intein 3' region	Saccharomyces cerevisiae VMA1	modified 3' region of the intein from t…	6180..6729	Fwd
DNA Sequence GGCCACGGTGGTATTCGCAATAATCTTAATACTGAGAATCCATTATGGGACGCTATTGTTGGCTTAGGATTCTTGAAGGACGGTGTCAAAAATATTCCTTCTTTCTTGTCTACGGACAATATCGGTACTCGTGAAACATTTCTTGCTGGTCTAATTGATTCTGATGGCTATGTTACTGATGAGCATGGTATTAAAGCAACAATAAAGACAATTCATACTTCTGTCAGAGATGGTTTGGTTTCCCTTGCTCGTTCTTTAGGCTTAGTAGTCTCGGTTAACGCAGAACCTGCTAAGGTTGACATGAATGTCACCAAACATAAAATTAGTTATGCTATTTATATGTCTGGTGGAGATGTTTTGCTTAACGTTCTTTCGAAGTGTGCCGGCTCTAAAAAATTCAGGCCTGCTCCCGCCGCTGCTTTTGCACGTGAGTGCCGCGGATTTTATTTCGAGTTACAAGAATTGAAGGAAGACGATTATTATGGGATTACTTTATCTGATGATTCTGATCATCAGTTTTTGCTTGGATCTCAGGTTGTTGTACAGAAC Protein Sequence GHGGIRNNLNTENPLWDAIVGLGFLKDGVKNIPSFLSTDNIGTRETFLAGLIDSDGYVTDEHGIKATIKTIHTSVRDGLVSLARSLGLVVSVNAEPAKVDMNVTKHKISYAIYMSGGDVLLNVLSKCAGSKKFRPAPAAAFARECRGFYFELQELKEDDYYGITLSDDSDHQFLLGSQVVVQN

promoter 3

Label	Gene	Product	Position	Strand
AmpR promoter	bla	—	13..118	Fwd
DNA Sequence CGCGGAACCCCTATTTGTTTATTTTTCTAAATACATTCAAATATGTATCCGCTCATGAGACAATAACCCTGATAAATGCTTCAATAATATTGAAAAAGGAAGAGT
lacI promoter	lacI	—	4432..4510	Rev
DNA Sequence GACACCATCGAATGGCGCAAAACCTTTCGCGGTATGGCATGATAGCGCCCGGAAGAGAGTCAATTCAGGGTGGTGAAT
T7 promoter	—	—	5066..5085	Fwd
DNA Sequence TAATACGACTCACTATAGG

protein_bind 1

Label	Gene	Product	Position	Strand
lac operator	—	—	5085..5110	Fwd
DNA Sequence GGAATTGTGAGCGGATAACAATTCC

rep_origin 2

Label	Gene	Product	Position	Strand
M13 ori	—	—	1020..1534	Rev
DNA Sequence ACGCGCCCTGTAGCGGCGCATTAAGCGCGGCGGGTGTGGTGGTTACGCGCAGCGTGACCGCTACACTTGCCAGCGCCCTAGCGCCCGCTCCTTTCGCTTTCTTCCCTTCCTTTCTCGCCACGTTCGCCGGCTTTCCCCGTCAAGCTCTAAATCGGGGGCTCCCTTTAGGGTTCCGATTTAGTGCTTTACGGCACCTCGACCCCAAAAAACTTGATTTGGGTGATGGTTCACGTAGTGGGCCATCGCCCTGATAGACGGTTTTTCGCCCTTTGACGTTGGAGTCCACGTTCTTTAATAGTGGACTCTTGTTCCAAACTGGAACAACACTCAACCCTATCTCGGGCTATTCTTTTGATTTATAAGGGATTTTGCCGATTTCGGCCTATTGGTTAAAAAATGAGCTGATTTAACAAAAATTTAACGCGAATTTTAACAAAATATTAACGTTTACAATTTAAATATTTGCTTATACAATCTTCCTGTTTTTGGGGCTTTTCTGATTATCAACCGGGGT
ori	—	—	1644..2233	Fwd
DNA Sequence TTGAGATCCTTTTTTTCTGCGCGTAATCTGCTGCTTGCAAACAAAAAAACCACCGCTACCAGCGGTGGTTTGTTTGCCGGATCAAGAGCTACCAACTCTTTTTCCGAAGGTAACTGGCTTCAGCAGAGCGCAGATACCAAATACTGTCCTTCTAGTGTAGCCGTAGTTAGGCCACCACTTCAAGAACTCTGTAGCACCGCCTACATACCTCGCTCTGCTAATCCTGTTACCAGTGGCTGCTGCCAGTGGCGATAAGTCGTGTCTTACCGGGTTGGACTCAAGACGATAGTTACCGGATAAGGCGCAGCGGTCGGGCTGAACGGGGGGTTCGTGCACACAGCCCAGCTTGGAGCGAACGACCTACACCGAACTGAGATACCTACAGCGTGAGCTATGAGAAAGCGCCACGCTTCCCGAAGGGAGAAAGGCGGACAGGTATCCGGTAAGCGGCAGGGTCGGAACAGGAGAGCGCACGAGGGAGCTTCCAGGGGGAAACGCCTGGTATCTTTATAGTCCTGTCGGGTTTCGCCACCTCTGACTTGAGCGTCGATTTTTGTGATGCTCGTCAGGGGGGCGGAGCCTATGGAAA

terminator 3

Label	Gene	Product	Position	Strand
rrnB T1 terminator	Escherichia coli rrnB	—	4662..4749	Fwd
DNA Sequence CAAATAAAACGAAAGGCTCAGTCGAAAGACTGGGCCTTTCGTTTTATCTGTTGTTTGTCGGTGAACGCTCTCCTGAGTAGGACAAAT
rrnB T1 terminator	Escherichia coli rrnB	—	4845..4932	Fwd
DNA Sequence CAAATAAAACGAAAGGCTCAGTCGAAAGACTGGGCCTTTCGTTTTATCTGTTGTTTGTCGGTGAACGCTCTCCTGAGTAGGACAAAT
T7 terminator	—	—	7005..7053	Fwd
DNA Sequence CTAGCATAACCCCTTGGGGCCTCTAAACGGGTCTTGAGGGGTTTTTTG