Prham-M.XhoI-M.MunI-M.Ocy1ORF8430P-p15A-aadA

Plasmid Information

Purpose

Methylase expression vector for 5RM assembly: expresses M.XhoI, M.MunI (MfeI) and M.Ocy1ORF8430P (SpeI) DNA methyltransferases in E. coli

Depositor

Ichiro Matsumura

Article

Matsumura et al Biotechniques. 2022 May;72(5):185-193. d

Promoter

rhamnose

Available Since

Feb. 22, 2022

Availability

Available

Length

5,777 bp

Topology

circular

Molecule Type

other DNA

CDS 2

Label	Gene	Product	Position	Strand
S-Tag	—	affinity and epitope tag derived from p…	126..171	Rev
DNA Sequence AAAGAAACCGCTGCTGCGAAATTTGAACGCCAGCACATGGACTCG Protein Sequence KETAAAKFERQHMDS
SmR	aadA	aminoglycoside adenylyltransferase (Mur…	4842..5634	Fwd
DNA Sequence ATGAGGGAAGCGGTGATCGCCGAAGTATCGACTCAACTATCAGAGGTAGTTGGCGTCATCGAGCGCCATCTCGAACCGACGTTGCTGGCCGTACATTTGTACGGCTCCGCAGTGGATGGCGGCCTGAAGCCACACAGTGATATTGATTTGCTGGTTACGGTGACCGTAAGGCTTGATGAAACAACGCGGCGAGCTTTGATCAACGACCTTTTGGAAACTTCGGCTTCCCCTGGAGAGAGCGAGATTCTCCGCGCTGTAGAAGTCACCATTGTTGTGCACGACGACATCATTCCGTGGCGTTATCCAGCTAAGCGCGAACTGCAATTTGGAGAATGGCAGCGCAATGACATTCTTGCAGGTATCTTCGAGCCAGCCACGATCGACATTGATCTGGCTATCTTGCTGACAAAAGCAAGAGAACATAGCGTTGCCTTGGTAGGTCCAGCGGCGGAGGAACTCTTTGATCCGGTTCCTGAACAGGATCTATTTGAGGCGCTAAATGAAACCTTAACGCTATGGAACTCGCCGCCCGACTGGGCTGGCGATGAGCGAAATGTAGTGCTTACGTTGTCCCGCATTTGGTACAGCGCAGTAACCGGCAAAATCGCGCCGAAGGATGTCGCTGCCGACTGGGCAATGGAGCGCCTGCCGGCCCAGTATCAGCCCGTCATACTTGAAGCTAGACAGGCTTATCTTGGACAAGAAGAAGATCGCTTGGCCTCGCGCGCAGATCAGTTGGAAGAATTTGTCCACTACGTGAAAGGCGAGATCACCAAGGTAGTCGGCAAATAA Protein Sequence MREAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPHSDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAKRELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLFEALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQPVILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK

RBS 3

Label	Gene	Product	Position	Strand
Shine-Dalgarno sequence	—	—	468..477	Fwd
DNA Sequence TAAGGAGGT
Shine-Dalgarno sequence	—	—	2211..2220	Fwd
DNA Sequence TAAGGAGGT
Shine-Dalgarno sequence	—	—	2934..2943	Fwd
DNA Sequence TAAGGAGGT

promoter 2

Label	Gene	Product	Position	Strand
rhaB promoter	—	—	330..449	Fwd
DNA Sequence CACCACAATTCAGCAAATTGTGAACATCATCACGTTCATCTTTCCCTGGTTGCCAATGGCCCATTTTCCTGTCAGTAACGAGAAGGTCGCGAATTGAGGCGCTTTTTAGACTGGTCGTA
AmpR promoter	bla	—	4750..4842	Fwd
DNA Sequence TTTGTTTATTTTTCTAAATACATTCAAATATGTATCCGCTCATGAGACAATAACCCTGATAAATGCTTCAATAATATTGAAAAAGGAAGAGT

rep_origin 1

Label	Gene	Product	Position	Strand
p15A ori	—	—	3937..4483	Fwd
DNA Sequence TTGAGATCGTTTTGGTCTGCGCGTAATCTCTTGCTCTGAAAACGAAAAAACCGCCTTGCAGGGCGGTTTTTCGAAGGTTCTCTGAGCTACCAACTCTTTGAACCGAGGTAACTGGCTTGGAGGAGCGCAGTCACCAAAACTTGTCCTTTCAGTTTAGCCTTAACCGGCGCATGACTTCAAGACTAACTCCTCTAAATCAATTACCAGTGGCTGCTGCCAGTGGTGCTTTTGCATGTCTTTCCGGGTTGGACTCAAGACGATAGTTACCGGATAAGGCGCAGCGGTCGGACTGAACGGGGGGTTCGTGCATACAGTCCAGCTTGGAGCGAACTGCCTACCCGGAACTGAGTGTCAGGCGTGGAATGAGACAAACGCGGCCATAACAGCGGAATGACACCGGTAAACCGAAAGGCAGGAACAGGAGAGCGCACGAGGGAGCCGCCAGGGGGAAACGCCTGGTATCTTTATAGTCCTGTCGGGTTTCGCCACCACTGATTTGAGCGTCAGATTTCGTGATGCTTGTCAGGGGGGCGGAGCCTATGGAAA

terminator 1

Label	Gene	Product	Position	Strand
T7 terminator	—	—	27..75	Rev
DNA Sequence CTAGCATAACCCCTTGGGGCCTCTAAACGGGTCTTGAGGGGTTTTTTG