Prham-M.XbaI-M.EcoRI-M.SalI-p15A-aadA

Plasmid Information

Purpose

Methylase expression vector for 5RM assembly: expresses M.XbaI, M.EcoRI and M.SalI 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

6,437 bp

Topology

circular

Molecule Type

other DNA

CDS 2

Label	Gene	Product	Position	Strand
SmR	aadA	aminoglycoside adenylyltransferase (Mur…	1573..2365	Fwd
DNA Sequence ATGAGGGAAGCGGTGATCGCCGAAGTATCGACTCAACTATCAGAGGTAGTTGGCGTCATCGAGCGCCATCTCGAACCGACGTTGCTGGCCGTACATTTGTACGGCTCCGCAGTGGATGGCGGCCTGAAGCCACACAGTGATATTGATTTGCTGGTTACGGTGACCGTAAGGCTTGATGAAACAACGCGGCGAGCTTTGATCAACGACCTTTTGGAAACTTCGGCTTCCCCTGGAGAGAGCGAGATTCTCCGCGCTGTAGAAGTCACCATTGTTGTGCACGACGACATCATTCCGTGGCGTTATCCAGCTAAGCGCGAACTGCAATTTGGAGAATGGCAGCGCAATGACATTCTTGCAGGTATCTTCGAGCCAGCCACGATCGACATTGATCTGGCTATCTTGCTGACAAAAGCAAGAGAACATAGCGTTGCCTTGGTAGGTCCAGCGGCGGAGGAACTCTTTGATCCGGTTCCTGAACAGGATCTATTTGAGGCGCTAAATGAAACCTTAACGCTATGGAACTCGCCGCCCGACTGGGCTGGCGATGAGCGAAATGTAGTGCTTACGTTGTCCCGCATTTGGTACAGCGCAGTAACCGGCAAAATCGCGCCGAAGGATGTCGCTGCCGACTGGGCAATGGAGCGCCTGCCGGCCCAGTATCAGCCCGTCATACTTGAAGCTAGACAGGCTTATCTTGGACAAGAAGAAGATCGCTTGGCCTCGCGCGCAGATCAGTTGGAAGAATTTGTCCACTACGTGAAAGGCGAGATCACCAAGGTAGTCGGCAAATAA Protein Sequence MREAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPHSDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAKRELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLFEALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQPVILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK
S-Tag	—	affinity and epitope tag derived from p…	2634..2679	Rev
DNA Sequence AAAGAAACCGCTGCTGCGAAATTTGAACGCCAGCACATGGACTCG Protein Sequence KETAAAKFERQHMDS

RBS 2

Label	Gene	Product	Position	Strand
Shine-Dalgarno sequence	—	—	4281..4290	Fwd
DNA Sequence TAAGGAGGT
Shine-Dalgarno sequence	—	—	5283..5292	Fwd
DNA Sequence TAAGGAGGT

promoter 2

Label	Gene	Product	Position	Strand
AmpR promoter	bla	—	1481..1573	Fwd
DNA Sequence TTTGTTTATTTTTCTAAATACATTCAAATATGTATCCGCTCATGAGACAATAACCCTGATAAATGCTTCAATAATATTGAAAAAGGAAGAGT
rhaB promoter	—	—	2838..2957	Fwd
DNA Sequence CACCACAATTCAGCAAATTGTGAACATCATCACGTTCATCTTTCCCTGGTTGCCAATGGCCCATTTTCCTGTCAGTAACGAGAAGGTCGCGAATTGAGGCGCTTTTTAGACTGGTCGTA

rep_origin 1

Label	Gene	Product	Position	Strand
p15A ori	—	—	668..1214	Fwd
DNA Sequence TTGAGATCGTTTTGGTCTGCGCGTAATCTCTTGCTCTGAAAACGAAAAAACCGCCTTGCAGGGCGGTTTTTCGAAGGTTCTCTGAGCTACCAACTCTTTGAACCGAGGTAACTGGCTTGGAGGAGCGCAGTCACCAAAACTTGTCCTTTCAGTTTAGCCTTAACCGGCGCATGACTTCAAGACTAACTCCTCTAAATCAATTACCAGTGGCTGCTGCCAGTGGTGCTTTTGCATGTCTTTCCGGGTTGGACTCAAGACGATAGTTACCGGATAAGGCGCAGCGGTCGGACTGAACGGGGGGTTCGTGCATACAGTCCAGCTTGGAGCGAACTGCCTACCCGGAACTGAGTGTCAGGCGTGGAATGAGACAAACGCGGCCATAACAGCGGAATGACACCGGTAAACCGAAAGGCAGGAACAGGAGAGCGCACGAGGGAGCCGCCAGGGGGAAACGCCTGGTATCTTTATAGTCCTGTCGGGTTTCGCCACCACTGATTTGAGCGTCAGATTTCGTGATGCTTGTCAGGGGGGCGGAGCCTATGGAAA

terminator 1

Label	Gene	Product	Position	Strand
T7 terminator	—	—	2535..2583	Rev
DNA Sequence CTAGCATAACCCCTTGGGGCCTCTAAACGGGTCTTGAGGGGTTTTTTG