pCIFR-6

227665 Synthetic Bacterial Expression

Plasmid Information

Purpose
Integration of msfGFP (or your Module Of Interest) in a random fashion via Tn5 insertion. The antibiotic cassette (Gm) 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
5,488 bp
Topology
circular
Molecule Type
other DNA
View on AddGene.org

Features

CDS 3
Label Gene Product Position Strand
AmpR bla beta-lactamase 0..5488 Fwd
DNA Sequence
ATGAGCATTCAGCATTTTCGTGTGGCGCTGATTCCGTTTTTTGCGGCGTTTTGCCTGCCGGTGTTTGCGCATCCGGAAACCCTGGTGAAAGTGAAAGATGCGGAAGATCAACTGGGTGCGCGCGTGGGCTATATTGAACTGGATCTGAACAGCGGCAAAATTCTGGAATCTTTTCGTCCGGAAGAACGTTTTCCGATGATGAGCACCTTTAAAGTGCTGCTGTGCGGTGCGGTTCTGAGCCGTGTGGATGCGGGCCAGGAACAACTGGGCCGTCGTATTCATTATAGCCAGAACGATCTGGTGGAATATAGCCCGGTGACCGAAAAACATCTGACCGATGGCATGACCGTGCGTGAACTGTGCAGCGCGGCGATTACCATGAGCGATAACACCGCGGCGAACCTGCTGCTGACGACCATTGGCGGTCCGAAAGAACTGACCGCGTTTCTGCATAACATGGGCGATCATGTGACCCGTCTGGATCGTTGGGAACCGGAACTGAACGAAGCGATTCCGAACGATGAACGTGATACCACCATGCCGGCAGCAATGGCGACCACCCTGCGTAAACTGCTGACGGGTGAGCTGCTGACCCTGGCAAGCCGCCAGCAACTGATTGATTGGATGGAAGCGGATAAAGTGGCGGGTCCGCTGCTGCGTAGCGCGCTGCCGGCTGGCTGGTTTATTGCGGATAAAAGCGGTGCGGGCGAACGTGGCAGCCGTGGCATTATTGCGGCGCTGGGCCCGGATGGTAAACCGAGCCGTATTGTGGTGATTTATACCACCGGCAGCCAGGCGACGATGGATGAACGTAACCGTCAGATTGCGGAAATTGGCGCGAGCCTGATTAAACATTGGTAA
Protein Sequence
MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYIELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYSPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRWEPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGASLIKHW
GmR aacC1 gentamycin acetyltransferase 2302..2836 Fwd
DNA Sequence
ATGTTACGCAGCAGCAACGATGTTACGCAGCAGGGCAGTCGCCCTAAAACAAAGTTAGGTGGCTCAAGTATGGGCATCATTCGCACATGTAGGCTCGGCCCTGACCAAGTCAAATCCATGCGGGCTGCTCTTGATCTTTTCGGTCGTGAGTTCGGAGACGTAGCCACCTACTCCCAACATCAGCCGGACTCCGATTACCTCGGGAACTTGCTCCGTAGTAAGACATTCATCGCGCTTGCTGCCTTCGACCAAGAAGCGGTTGTTGGCGCTCTCGCGGCTTACGTTCTGCCCAAGTTTGAGCAGCCGCGTAGTGAGATCTATATCTATGATCTCGCAGTCTCCGGAGAGCACCGGAGGCAGGGCATTGCCACCGCGCTCATCAATCTCCTCAAGCATGAGGCCAACGCGCTTGGTGCTTATGTGATCTACGTGCAAGCAGATTACGGTGACGATCCCGCAGTGGCTCTCTATACAAAGTTGGGCATACGGGAAGAAGTGATGCACTTTGATATCGACCCAAGTACCGCCACCTAA
Protein Sequence
MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALDLFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRSEIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIREEVMHFDIDPSTAT
Tn5 transposase — transposase from the bacterial Tn5 tran… 3257..4685 Fwd
DNA Sequence
ATGATTACCAGTGCACTGCATCGTGCGGCGGATTGGGCGAAAAGCGTGTTTTCTAGTGCTGCGCTGGGTGATCCGCGTCGTACCGCGCGTCTGGTGAATGTTGCGGCGCAACTGGCCAAATATAGCGGCAAAAGCATTACCATTAGCAGCGAAGGCAGCAAAGCCATGCAGGAAGGCGCGTATCGTTTTATTCGTAATCCGAACGTGAGCGCGGAAGCGATTCGTAAAGCGGGTGCCATGCAGACCGTGAAACTGGCCCAGGAATTTCCGGAACTGCTGGCAATTGAAGATACCACCTCTCTGAGCTATCGTCATCAGGTGGCGGAAGAACTGGGCAAACTGGGTAGCATTCAGGATAAAAGCCGTGGTTGGTGGGTGCATAGCGTGCTGCTGCTGGAAGCGACCACCTTTCGTACCGTGGGCCTGCTGCATCAAGAATGGTGGATGCGTCCGGATGATCCGGCGGATGCGGATGAAAAAGAAAGCGGCAAATGGCTGGCCGCTGCTGCAACTTCGCGTCTGAGAATGGGCAGCATGATGAGCAACGTGATTGCGGTGTGCGATCGTGAAGCGGATATTCATGCGTATCTGCAAGATAAACTGGCCCATAACGAACGTTTTGTGGTGCGTAGCAAACATCCGCGTAAAGATGTGGAAAGCGGCCTGTATCTGTATGATCACCTGAAAAACCAGCCGGAACTGGGCGGCTATCAGATTAGCATTCCGCAGAAAGGCGTGGTGGATAAACGTGGCAAACGTAAAAACCGTCCGGCGCGTAAAGCGAGCCTGAGCCTGCGTAGCGGCCGTATTACCCTGAAACAGGGCAACATTACCCTGAACGCGGTGCTGGCCGAAGAAATTAATCCGCCGAAAGGCGAAACCCCGCTGAAATGGCTGCTGCTGACCAGCGAGCCGGTGGAAAGTCTGGCCCAAGCGCTGCGTGTGATTGATATTTATACCCATCGTTGGCGCATTGAAGAATTTCACAAAGCGTGGAAAACGGGTGCGGGTGCGGAACGTCAGCGTATGGAAGAACCGGATAACCTGGAACGTATGGTGAGCATTCTGAGCTTTGTGGCGGTGCGTCTGCTGCAACTGCGTGAATCTTTTACTCCGCCGCAAGCACTGCGTGCGCAGGGCCTGCTGAAAGAAGCGGAACACGTTGAAAGCCAGAGCGCGGAAACCGTGCTGACCCCGGATGAATGCCAACTGCTGGGCTATCTGGATAAAGGCAAACGCAAACGCAAAGAAAAAGCGGGCAGCCTGCAATGGGCGTATATGGCGATTGCGCGTCTGGGCGGCTTTATGGATAGCAAACGTACCGGCATTGCGAGCTGGGGTGCGCTGTGGGAAGGTTGGGAAGCGCTGCAAAGCAAACTGGATGGCTTTCTGGCCGCGAAAGACCTGATGGCGCAGGGCATTAAAATC
Protein Sequence
MITSALHRAADWAKSVFSSAALGDPRRTARLVNVAAQLAKYSGKSITISSEGSKAMQEGAYRFIRNPNVSAEAIRKAGAMQTVKLAQEFPELLAIEDTTSLSYRHQVAEELGKLGSIQDKSRGWWVHSVLLLEATTFRTVGLLHQEWWMRPDDPADADEKESGKWLAAAATSRLRMGSMMSNVIAVCDREADIHAYLQDKLAHNERFVVRSKHPRKDVESGLYLYDHLKNQPELGGYQISIPQKGVVDKRGKRKNRPARKASLSLRSGRITLKQGNITLNAVLAEEINPPKGETPLKWLLLTSEPVESLAQALRVIDIYTHRWRIEEFHKAWKTGAGAERQRMEEPDNLERMVSILSFVAVRLLQLRESFTPPQALRAQGLLKEAEHVESQSAETVLTPDECQLLGYLDKGKRKRKEKAGSLQWAYMAIARLGGFMDSKRTGIASWGALWEGWEALQSKLDGFLAAKDLMAQGIKI
RBS 1
Label Gene Product Position Strand
Shine-Dalgarno sequence — — 1225..1234 Fwd
DNA Sequence
TAAGGAGGT
promoter 2
Label Gene Product Position Strand
Pc promoter intI1 (promoter lies within the coding sequence) — 2085..2114 Fwd
DNA Sequence
TTGACATAAGCCTGTTCGGTTCGTAAACT
AmpR promoter bla — 4741..4833 Fwd
DNA Sequence
TTTGTTTATTTTTCTAAATACATTCAAATATGTATCCGCTCATGAGACAATAACCCTGATAAATGCTTCAATAATATTGAAAAAGGAAGAGT
protein_bind 2
Label Gene Product Position Strand
Bxb1 attP — — 2027..2075 Fwd
DNA Sequence
GGTTTGTCTGGTCAACCACCGCGGTCTCAGTGGTGTACGGTACAAACC
Bxb1 attB — — 2986..3024 Fwd
DNA Sequence
GGCTTGTCGACGACGGCGGTCTCCGTCGTCAGGATCAT
rep_origin 1
Label Gene Product Position Strand
R6K gamma ori — — 515..904 Fwd
DNA Sequence
GATCTGAAGATCAGCAGTTCAACCTGTTGATAGTACGTACTAAGCTCTCATGTTTCACGTACTAAGCTCTCATGTTTAACGTACTAAGCTCTCATGTTTAACGAACTAAACCCTCATGGCTAACGTACTAAGCTCTCATGGCTAACGTACTAAGCTCTCATGTTTCACGTACTAAGCTCTCATGTTTGAACAATAAAATTAATATAAATCAGCAACTTAAATAGCCTCTAAGGTTTTAAGTTTTATAAGAAAAAAAAGAATATATAAGGCTTTTAAAGCCTTTAAGGTTTAACGGTTGTGGACAACAAGCCAGGGATGTAACGCACTGAGAAGCCCTTAGAGCCTCTCAAAGCAATTTTGAGTGACACAGGAACACTTAACGGCTGACA
terminator 3
Label Gene Product Position Strand
fd terminator — — 209..249 Fwd
DNA Sequence
GATACAATTAAAGGCTCCTTTTGGAGCCTTTTTTTTTGGA
rrnB T1 terminator Escherichia coli rrnB — 934..1021 Rev
DNA Sequence
CAAATAAAACGAAAGGCTCAGTCGAAAGACTGGGCCTTTCGTTTTATCTGTTGTTTGTCGGTGAACGCTCTCCTGAGTAGGACAAAT
lambda t0 terminator — — 3058..3153 Fwd
DNA Sequence
GACTCCTGTTGATAGATCCAGTAATGACCTCAGAACTCCATCTGGATTTGTTCAGAACGCTCGGTTGCCGCCGGGCGTTTTTTATTGGTGAGAAT