pCIFR-2

227663 Synthetic Bacterial Expression

Plasmid Information

Purpose
Integration of msfGFP (or your Module Of Interest) in a random fashion via Tn5 insertion. The antibiotic cassette (Km) used for selecting for the integration can be removed in a later stage with pFNC.
Depositor
Pablo Ivan Nikel
Article
Federici et al Metab Eng. 2025 Jan 6;88:180-195. doi: 1
Insert
msfGFP
Promoter
p14G-BCD2
Available Since
Jan. 22, 2025
Availability
Available

Sequence Information

Length
5,600 bp
Topology
circular
Molecule Type
other DNA

Features

CDS 3
Label Gene Product Position Strand
KanR aph(3')-Ia aminoglycoside phosphotransferase 1160..1976 Fwd
DNA Sequence
ATGAGCCATATTCAGCGTGAAACGAGCTGTAGCCGTCCGCGTCTGAACAGCAACATGGATGCGGATCTGTATGGCTATAAATGGGCGCGTGATAACGTGGGTCAGAGCGGCGCGACCATTTATCGTCTGTATGGCAAACCGGATGCGCCGGAACTGTTTCTGAAACATGGCAAAGGCAGCGTGGCGAACGATGTGACCGATGAAATGGTGCGTCTGAACTGGCTGACCGAATTTATGCCGCTGCCGACCATTAAACATTTTATTCGCACCCCGGATGATGCGTGGCTGCTGACCACCGCGATTCCGGGCAAAACCGCGTTTCAGGTGCTGGAAGAATATCCGGATAGCGGCGAAAACATTGTGGATGCGCTGGCCGTGTTTCTGCGTCGTCTGCATAGCATTCCGGTGTGCAACTGCCCGTTTAACAGCGATCGTGTGTTTCGTCTGGCCCAGGCGCAGAGCCGTATGAACAACGGCCTGGTGGATGCGAGCGATTTTGATGATGAACGTAACGGCTGGCCGGTGGAACAGGTGTGGAAAGAAATGCATAAACTGCTGCCGTTTAGCCCGGATAGCGTGGTGACCCACGGCGATTTTAGCCTGGATAACCTGATTTTCGATGAAGGCAAACTGATTGGCTGCATTGATGTGGGCCGTGTGGGCATTGCGGATCGTTATCAGGATCTGGCCATTCTGTGGAACTGCCTGGGCGAATTTAGCCCGAGCCTGCAAAAACGTCTGTTTCAGAAATATGGCATTGATAATCCGGATATGAACAAACTGCAATTTCATCTGATGCTGGATGAATTTTTCTAA
Protein Sequence
MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF
Tn5 transposase transposase from the bacterial Tn5 tran… 2343..3771 Fwd
DNA Sequence
ATGATTACCAGTGCACTGCATCGTGCGGCGGATTGGGCGAAAAGCGTGTTTTCTAGTGCTGCGCTGGGTGATCCGCGTCGTACCGCGCGTCTGGTGAATGTTGCGGCGCAACTGGCCAAATATAGCGGCAAAAGCATTACCATTAGCAGCGAAGGCAGCAAAGCCATGCAGGAAGGCGCGTATCGTTTTATTCGTAATCCGAACGTGAGCGCGGAAGCGATTCGTAAAGCGGGTGCCATGCAGACCGTGAAACTGGCCCAGGAATTTCCGGAACTGCTGGCAATTGAAGATACCACCTCTCTGAGCTATCGTCATCAGGTGGCGGAAGAACTGGGCAAACTGGGTAGCATTCAGGATAAAAGCCGTGGTTGGTGGGTGCATAGCGTGCTGCTGCTGGAAGCGACCACCTTTCGTACCGTGGGCCTGCTGCATCAAGAATGGTGGATGCGTCCGGATGATCCGGCGGATGCGGATGAAAAAGAAAGCGGCAAATGGCTGGCCGCTGCTGCAACTTCGCGTCTGAGAATGGGCAGCATGATGAGCAACGTGATTGCGGTGTGCGATCGTGAAGCGGATATTCATGCGTATCTGCAAGATAAACTGGCCCATAACGAACGTTTTGTGGTGCGTAGCAAACATCCGCGTAAAGATGTGGAAAGCGGCCTGTATCTGTATGATCACCTGAAAAACCAGCCGGAACTGGGCGGCTATCAGATTAGCATTCCGCAGAAAGGCGTGGTGGATAAACGTGGCAAACGTAAAAACCGTCCGGCGCGTAAAGCGAGCCTGAGCCTGCGTAGCGGCCGTATTACCCTGAAACAGGGCAACATTACCCTGAACGCGGTGCTGGCCGAAGAAATTAATCCGCCGAAAGGCGAAACCCCGCTGAAATGGCTGCTGCTGACCAGCGAGCCGGTGGAAAGTCTGGCCCAAGCGCTGCGTGTGATTGATATTTATACCCATCGTTGGCGCATTGAAGAATTTCACAAAGCGTGGAAAACGGGTGCGGGTGCGGAACGTCAGCGTATGGAAGAACCGGATAACCTGGAACGTATGGTGAGCATTCTGAGCTTTGTGGCGGTGCGTCTGCTGCAACTGCGTGAATCTTTTACTCCGCCGCAAGCACTGCGTGCGCAGGGCCTGCTGAAAGAAGCGGAACACGTTGAAAGCCAGAGCGCGGAAACCGTGCTGACCCCGGATGAATGCCAACTGCTGGGCTATCTGGATAAAGGCAAACGCAAACGCAAAGAAAAAGCGGGCAGCCTGCAATGGGCGTATATGGCGATTGCGCGTCTGGGCGGCTTTATGGATAGCAAACGTACCGGCATTGCGAGCTGGGGTGCGCTGTGGGAAGGTTGGGAAGCGCTGCAAAGCAAACTGGATGGCTTTCTGGCCGCGAAAGACCTGATGGCGCAGGGCATTAAAATC
Protein Sequence
MITSALHRAADWAKSVFSSAALGDPRRTARLVNVAAQLAKYSGKSITISSEGSKAMQEGAYRFIRNPNVSAEAIRKAGAMQTVKLAQEFPELLAIEDTTSLSYRHQVAEELGKLGSIQDKSRGWWVHSVLLLEATTFRTVGLLHQEWWMRPDDPADADEKESGKWLAAAATSRLRMGSMMSNVIAVCDREADIHAYLQDKLAHNERFVVRSKHPRKDVESGLYLYDHLKNQPELGGYQISIPQKGVVDKRGKRKNRPARKASLSLRSGRITLKQGNITLNAVLAEEINPPKGETPLKWLLLTSEPVESLAQALRVIDIYTHRWRIEEFHKAWKTGAGAERQRMEEPDNLERMVSILSFVAVRLLQLRESFTPPQALRAQGLLKEAEHVESQSAETVLTPDECQLLGYLDKGKRKRKEKAGSLQWAYMAIARLGGFMDSKRTGIASWGALWEGWEALQSKLDGFLAAKDLMAQGIKI
AmpR bla beta-lactamase 3919..4780 Fwd
DNA Sequence
ATGAGCATTCAGCATTTTCGTGTGGCGCTGATTCCGTTTTTTGCGGCGTTTTGCCTGCCGGTGTTTGCGCATCCGGAAACCCTGGTGAAAGTGAAAGATGCGGAAGATCAACTGGGTGCGCGCGTGGGCTATATTGAACTGGATCTGAACAGCGGCAAAATTCTGGAATCTTTTCGTCCGGAAGAACGTTTTCCGATGATGAGCACCTTTAAAGTGCTGCTGTGCGGTGCGGTTCTGAGCCGTGTGGATGCGGGCCAGGAACAACTGGGCCGTCGTATTCATTATAGCCAGAACGATCTGGTGGAATATAGCCCGGTGACCGAAAAACATCTGACCGATGGCATGACCGTGCGTGAACTGTGCAGCGCGGCGATTACCATGAGCGATAACACCGCGGCGAACCTGCTGCTGACGACCATTGGCGGTCCGAAAGAACTGACCGCGTTTCTGCATAACATGGGCGATCATGTGACCCGTCTGGATCGTTGGGAACCGGAACTGAACGAAGCGATTCCGAACGATGAACGTGATACCACCATGCCGGCAGCAATGGCGACCACCCTGCGTAAACTGCTGACGGGTGAGCTGCTGACCCTGGCAAGCCGCCAGCAACTGATTGATTGGATGGAAGCGGATAAAGTGGCGGGTCCGCTGCTGCGTAGCGCGCTGCCGGCTGGCTGGTTTATTGCGGATAAAAGCGGTGCGGGCGAACGTGGCAGCCGTGGCATTATTGCGGCGCTGGGCCCGGATGGTAAACCGAGCCGTATTGTGGTGATTTATACCACCGGCAGCCAGGCGACGATGGATGAACGTAACCGTCAGATTGCGGAAATTGGCGCGAGCCTGATTAAACATTGGTAA
Protein Sequence
MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYIELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYSPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRWEPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGASLIKHW
RBS 1
Label Gene Product Position Strand
Shine-Dalgarno sequence 199..208 Fwd
DNA Sequence
TAAGGAGGT
promoter 1
Label Gene Product Position Strand
AmpR promoter bla 3827..3919 Fwd
DNA Sequence
TTTGTTTATTTTTCTAAATACATTCAAATATGTATCCGCTCATGAGACAATAACCCTGATAAATGCTTCAATAATATTGAAAAAGGAAGAGT
protein_bind 2
Label Gene Product Position Strand
Bxb1 attP 1001..1049 Fwd
DNA Sequence
GGTTTGTCTGGTCAACCACCGCGGTCTCAGTGGTGTACGGTACAAACC
Bxb1 attB 2072..2110 Fwd
DNA Sequence
GGCTTGTCGACGACGGCGGTCTCCGTCGTCAGGATCAT
rep_origin 1
Label Gene Product Position Strand
R6K gamma ori 5089..5478 Fwd
DNA Sequence
GATCTGAAGATCAGCAGTTCAACCTGTTGATAGTACGTACTAAGCTCTCATGTTTCACGTACTAAGCTCTCATGTTTAACGTACTAAGCTCTCATGTTTAACGAACTAAACCCTCATGGCTAACGTACTAAGCTCTCATGGCTAACGTACTAAGCTCTCATGTTTCACGTACTAAGCTCTCATGTTTGAACAATAAAATTAATATAAATCAGCAACTTAAATAGCCTCTAAGGTTTTAAGTTTTATAAGAAAAAAAAGAATATATAAGGCTTTTAAAGCCTTTAAGGTTTAACGGTTGTGGACAACAAGCCAGGGATGTAACGCACTGAGAAGCCCTTAGAGCCTCTCAAAGCAATTTTGAGTGACACAGGAACACTTAACGGCTGACA
terminator 3
Label Gene Product Position Strand
lambda t0 terminator 2144..2239 Fwd
DNA Sequence
GACTCCTGTTGATAGATCCAGTAATGACCTCAGAACTCCATCTGGATTTGTTCAGAACGCTCGGTTGCCGCCGGGCGTTTTTTATTGGTGAGAAT
fd terminator 4783..4823 Fwd
DNA Sequence
GATACAATTAAAGGCTCCTTTTGGAGCCTTTTTTTTTGGA
rrnB T1 terminator Escherichia coli rrnB 5508..5595 Rev
DNA Sequence
CAAATAAAACGAAAGGCTCAGTCGAAAGACTGGGCCTTTCGTTTTATCTGTTGTTTGTCGGTGAACGCTCTCCTGAGTAGGACAAAT