pMamCol1

215184 Synthetic Bacterial Expression

Plasmid Information

Purpose
Allows for stable expression of recombinant (tagged/untagged) proteins in AAVS1 locus of Human cells and transient Protein Expression in E. coli
Depositor
Krishna Kishore Inampudi
Insert
tsPurple
Tags
SNAC-StrepII tag
Promoter
TetON 3G Bidirectional for Mammalian and Trc with…
Available Since
April 19, 2024
Availability
Available

Sequence Information

Length
14,602 bp
Topology
circular
Molecule Type
other DNA

Features

CDS 11
Label Gene Product Position Strand
SV40 NLS nuclear localization signal of SV40 (si… 1348..1369 Rev
DNA Sequence
CCTAAGAAGAAGCGGAAGGTG
Protein Sequence
PKKKRKV
enterokinase site enterokinase recognition and cleavage s… 2428..2443 Rev
DNA Sequence
GATGACGACGACAAG
Protein Sequence
DDDDK
Cas9(N) N-terminal portion of Streptococcus pyo… 3769..5488 Rev
DNA Sequence
ATGGACAAGAAGTACTCTATCGGCCTGGATATCGGCACAAACAGTGTGGGCTGGGCCGTGATTACCGACGAATACAAGGTCCCCTCTAAGAAGTTCAAGGTTCTGGGAAACACAGACAGACACTCTATCAAGAAAAACCTGATCGGCGCATTGCTGTTCGACTCCGGCGAGACGGCCGAAGCCACCAGACTGAAGAGAACTGCTCGGCGGAGATACACAAGACGGAAGAATAGAATTTGCTACCTGCAGGAAATCTTCAGCAACGAGATGGCCAAGGTGGACGATAGCTTCTTCCACAGACTGGAGGAATCTTTCCTAGTCGAGGAAGACAAGAAGCACGAGCGCCACCCAATCTTCGGCAATATCGTGGACGAAGTGGCCTACCACGAGAAATACCCTACAATCTACCACCTGAGAAAGAAGCTCGTCGATAGCACCGACAAAGCAGATCTGAGACTGATCTACCTGGCCCTTGCCCACATGATTAAGTTCCGGGGTCACTTTCTGATCGAGGGCGACCTAAACCCTGATAACAGCGACGTGGACAAGCTATTCATCCAGCTGGTTCAGACCTACAATCAGCTGTTTGAGGAGAACCCCATCAACGCTTCCGGAGTGGACGCCAAGGCTATTCTCTCTGCCAGACTGTCAAAGAGCAGAAGACTTGAGAATTTGATCGCTCAGCTCCCTGGCGAGAAGAAGAACGGACTGTTTGGCAACCTTATCGCCCTGAGCCTGGGCCTGACCCCCAACTTCAAGAGCAATTTCGACCTGGCAGAAGACGCTAAGCTGCAGCTGTCTAAAGATACCTACGACGACGACCTGGACAACCTGCTGGCCCAGATCGGAGATCAGTACGCCGACCTGTTCCTGGCTGCCAAGAACCTGAGCGACGCTATCTTGCTGAGCGATATCCTGAGAGTGAACACCGAGATCACCAAAGCCCCTCTGAGCGCCTCTATGATTAAGCGGTACGACGAGCACCACCAGGATCTGACGCTGCTGAAAGCCCTGGTCAGACAGCAACTGCCAGAAAAGTATAAGGAAATCTTCTTTGACCAGAGCAAGAACGGCTACGCCGGATACATCGACGGCGGCGCCTCCCAGGAGGAGTTCTACAAATTCATCAAGCCTATCCTGGAAAAGATGGATGGCACCGAGGAACTGCTGGTGAAGCTGAACAGGGAGGACCTGCTGAGAAAGCAGAGAACTTTTGATAACGGTTCTATCCCCCACCAGATCCACCTGGGCGAACTCCACGCAATCCTGAGACGGCAGGAGGATTTTTACCCGTTCCTGAAAGATAACAGAGAGAAAATCGAAAAAATTCTGACCTTCAGAATCCCCTACTACGTGGGCCCTCTGGCCAGAGGCAACAGCAGATTCGCCTGGATGACCCGGAAGAGCGAAGAGACAATCACCCCTTGGAATTTCGAGGAAGTCGTGGATAAGGGCGCCTCTGCCCAGAGCTTTATCGAGCGGATGACCAACTTCGACAAGAACCTCCCCAACGAGAAAGTGCTGCCTAAGCACAGCCTCCTGTACGAGTACTTCACAGTGTATAACGAGCTGACAAAGGTCAAGTACGTGACCGAAGGAATGAGAAAGCCCGCCTTCCTGTCCGGTGAGCAGAAGAAAGCTATCGTGGATCTGTTGTTCAAGACCAACAGAAAGGTGACCGTTAAGCAGCTGAAGGAGGATTACTTTAAGAAGATCGAG
Protein Sequence
MDKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKNLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESFLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKFRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQIGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLRKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGNSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYFTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIE
Strep-Tag II peptide that binds Strep-Tactin(R), an … 7626..7650 Rev
DNA Sequence
TGGTCCCACCCTCAGTTTGAGAAG
Protein Sequence
WSHPQFEK
SNAC tag tag for protein cleavage using Ni2+ ion… 7659..7674 Rev
DNA Sequence
GGCTCTCACCACTGG
Protein Sequence
GSHHW
T2A 2A peptide from Thosea asigna virus cap… 9687..9741 Fwd
DNA Sequence
GAGGGCAGAGGCAGTCTGCTGACTTGCGGCGATGTGGAGGAAAATCCTGGACCT
Protein Sequence
EGRGSLLTCGDVEENPGP
Tet-On(R) 3G modified rtTA protein that binds tightl… 9741..10488 Fwd
DNA Sequence
ATGAGCAGACTGGACAAGTCCAAAGTGATTAACAGCGCCCTTGAACTGCTGAACGGCGTTGGAATCGAAGGCCTGACAACCCGGAAGCTGGCCCAGAAGCTGGGGGTGGAGCAGCCTACACTGTACTGGCACGTGAAGAACAAGAGAGCACTCCTGGACGCCCTGCCTATCGAGATGCTGGACAGACACCACACACACAGCTGCCCTCTGGAAGGCGAGAGCTGGCAGGACTTCCTGCGGAACAACGCCAAGTCTTATAGGTGCGCCCTGCTGAGCCACCGGGACGGCGCCAAGGTGCACCTGGGCACCAGACCTACCGAGAAACAGTACGAGACACTGGAGAACCAGCTGGCCTTCCTGTGCCAGCAAGGCTTCTCCCTGGAGAACGCCCTGTACGCCCTGAGCGCCGTGGGACACTTCACCCTGGGCTGTGTGCTGGAAGAGCAGGAGCACCAAGTGGCCAAAGAGGAGAGGGAAACCCCAACCACCGACTCTATGCCTCCTCTGCTGAAACAGGCTATCGAACTGTTCGATAGACAGGGCGCCGAGCCTGCCTTTCTGTTCGGCCTCGAGCTGATTATCTGTGGCCTGGAAAAGCAGCTGAAGTGCGAGTCTGGCGGACCCACCGACGCTCTGGACGATTTTGATCTGGATATGCTGCCTGCCGACGCTCTGGACGATTTTGATCTGGATATGCTGCCAGCCGACGCCCTCGACGACTTCGACCTGGATATGTTACCTGGCTGA
Protein Sequence
MSRLDKSKVINSALELLNGVGIEGLTTRKLAQKLGVEQPTLYWHVKNKRALLDALPIEMLDRHHTHSCPLEGESWQDFLRNNAKSYRCALLSHRDGAKVHLGTRPTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEEQEHQVAKEERETPTTDSMPPLLKQAIELFDRQGAEPAFLFGLELIICGLEKQLKCESGGPTDALDDFDLDMLPADALDDFDLDMLPADALDDFDLDMLPG
HygR aph(4)-Ia aminoglycoside phosphotransferase from … 10730..11756 Rev
DNA Sequence
ATGAAGAAACCGGAACTGACAGCCACAAGCGTTGAAAAATTTCTGATCGAGAAGTTCGATAGCGTTAGCGATCTGATGCAGCTGAGCGAAGGCGAAGAGAGCCGGGCGTTTAGCTTCGACGTCGGCGGAAGAGGCTACGTGCTGAGAGTGAACAGCTGCGCCGACGGTTTTTATAAAGATCGTTACGTGTATCGTCACTTTGCAAGTGCAGCCCTGCCCATCCCCGAGGTTCTGGATATTGGCGAGTTCAGCGAGTCTCTGACCTACTGCATCAGCCGTCGTGCACAGGGTGTTACACTCCAGGATCTGCCGGAAACCGAGCTGCCTGCTGTTCTGCAGCCGGTTGCCGAAGCAATGGACGCGATTGCCGCAGCTGATCTGAGCCAGACCAGCGGGTTTGGTCCGTTTGGACCTCAGGGCATCGGCCAGTACACCACCTGGCGTGATTTTATTTGTGCGATTGCAGATCCTCACGTGTACCACTGGCAGACCGTGATGGACGATACAGTTTCCGCCAGCGTGGCTCAGGCACTGGACGAACTGATGCTGTGGGCAGAAGATTGTCCTGAAGTGCGGCACCTGGTTCACGCCGATTTTGGTAGTAATAACGTGCTGACCGACAACGGCCGTATTACCGCCGTGATCGATTGGAGCGAGGCGATGTTCGGCGACTCCCAATACGAAGTTGCAAATATTTTCTTCTGGCGGCCGTGGCTGGCCTGTATGGAACAGCAGACACGCTATTTTGAAAGACGTCACCCGGAACTGGCAGGTAGCCCTAGACTGCGTGCCTACATGCTCCGGATCGGACTGGATCAGCTGTATCAGAGCCTGGTTGACGGCAACTTCGACGACGCCGCGTGGGCCCAAGGTCGCTGCGACGCGATCGTGCGCAGCGGCGCAGGCACCGTGGGCCGTACCCAGATCGCCAGAAGAAGCGCCGCCGTTTGGACCGACGGTTGTGTGGAAGTCCTGGCAGATAGCGGTAATAGAAGACCTAGCACCAGACCTCGTGCAAAAGAATAA
Protein Sequence
MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGYVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQTVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFGDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGNFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE
T2A 2A peptide from Thosea asigna virus cap… 11768..11822 Rev
DNA Sequence
GAGGGTCGTGGTAGCCTGCTGACCTGTGGTGACGTTGAGGAAAATCCGGGTCCA
Protein Sequence
EGRGSLLTCGDVEENPGP
Factor Xa site Factor Xa recognition and cleavage site 11813..11825 Rev
DNA Sequence
ATAGAGGGTCGT
Protein Sequence
IEGR
KanR aph(3')-Ia aminoglycoside phosphotransferase 13679..14489 Rev
DNA Sequence
ATGAGCCATATTCAACGGGAAACGTCGAGGCCGCGATTAAATTCCAACATGGATGCTGATTTATATGGGTATAAATGGGCTCGCGATAATGTCGGGCAATCAGGTGCGACAATCTATCGCTTGTATGGGAAGCCCGATGCGCCAGAGTTGTTTCTGAAACATGGCAAAGGTAGCGTTGCCAATGATGTTACAGATGAGATGGTCAGACTAAACTGGCTGACGGAATTTATGCCTCTTCCGACCATCAAGCATTTTATCCGTACTCCTGATGATGCATGGTTACTCACCACTGCGATCCCCGGAAAAACAGCATTCCAGGTATTAGAAGAATATCCTGATTCAGGTGAAAATATTGTTGATGCGCTGGCAGTGTTCCTGCGCCGGTTGCATTCGATTCCTGTTTGTAATTGTCCTTTTAACAGCGATCGCGTATTTCGCCTCGCTCAGGCGCAATCACGAATGAATAACGGTTTGGTTGATGCGAGTGATTTTGATGACGAGCGTAATGGCTGGCCTGTTGAACAAGTCTGGAAAGAAATGCATAAACTTTTGCCATTCTCACCGGATTCAGTCGTCACTCATGGTGATTTCTCACTTGATAACCTTATTTTTGACGAGGGGAAATTAATAGGTTGTATTGATGTTGGACGAGTCGGAATCGCAGACCGATACCAGGATCTTGCCATCCTATGGAACTGCCTCGGTGAGTTTTCTCCTTCATTACAGAAACGGCTTTTTCAAAAATATGGTATTGATAATCCTGATATGAATAAATTGCAGTTTCATTTGATGCTCGATGAGTTTTTCTAA
Protein Sequence
MSHIQRETSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF
intron 1
Label Gene Product Position Strand
hybrid intron 5505..5733 Fwd
DNA Sequence
CTGAAAAAAAGTGATTTCAGGCTCGTGCTCCAGGTAATTAAACTTAAATACCCCACCAACCAACCATCCCTTAAACCCTTACCTCTTGCTCAGCTAATTACAGCCCGGAGGAGAAGGGCCGTCCCGCCCGCTCACCTGTGGGAGTAACGCGGTCAGTCAGAGCCGGGGCGGGCGGCGCGAGGCGGCGGCGGAGCGGGGCACGGGGCGAAGGCAGCGCGCAGCGACTCC
polyA_signal 2
Label Gene Product Position Strand
HSV TK poly(A) signal 385..434 Fwd
DNA Sequence
GAACAAACGACCCAACACCCGTGCGTTTTATTCTGTCTTTTTATTGCCG
SV40 poly(A) signal 10494..10576 Fwd
DNA Sequence
AACTTGTTTATTGCAGCTTATATTGGTTACAAATAAAGCAATAGCATCACAAATTTCACAAATAAAGCATTTTTTTCACTGC
promoter 2
Label Gene Product Position Strand
chicken beta-actin promoter 5733..6010 Rev
DNA Sequence
TCGAGGTGAGCCCCACGTTCTGCTTCACTCTCCCCACCTCCCCCCCCTCCCCACCCCCAATTTTGTATTTATTTATTTTTTAATTATTTTGTGCAGCGTTGGGGGCGGGGGGGGGGGGGGGCGCGCGCCAGGCGGGGCGGGGCGGGGCGAGGGGCGGGGCGGGGCGAGGCGGAGAGGTGCGGCGGCAGCCAATCAGAGCGGCGCGCTCCGAAAGTTTCCTTTTCTGGCGAGGCGGCGGCGGCGGCGGCCCTATAAAAAGCGAAGCGCGCGGCGGGCG
TRE3G BI promoter 8498..8950 Fwd
DNA Sequence
TTTACGAGGGTAGGAAGTGGTACGGAAAGTTGGTATAAGACAAAAGTGTTGTGGAATTGCTCCAGGCGATCTGACGGTTCACTAAACTAGCTCTGCTTTTATAGGCGCCCACCGTACACGCCTAAACGTAATACGTTCTCTATCACTGATAGGGAGTAAACTGGATATACGTTCTCTATCACTGATAGGGAGTAAACTGTAGATACGTTCTCTATCACTGATAGGGAGTAAACTGGTCTTACGTTCTCTATCACTGATAGGGAGTAAACTCCTTATACGTTCTCTATCACTGATAGGGAGTAAAGTCTGCAAACGTTCTCTATCACTGATAGGGAGTAAACTCTTGATACGTTCTCTATCACTGATAGGGAGTAAACTCGAGTATGTCGAGGTAGGCGTGTACGGTGGGAGGTCTATATAAGCAGAGCTTGTTTAGTGAACCGTCAGATCGC
protein_bind 8
Label Gene Product Position Strand
lac operator 8428..8445 Fwd
DNA Sequence
TTGTTATCCGCTCACAA
tet operator tetO 8634..8653 Fwd
DNA Sequence
TCTCTATCACTGATAGGGA
tet operator tetO 8670..8689 Fwd
DNA Sequence
TCTCTATCACTGATAGGGA
tet operator tetO 8706..8725 Fwd
DNA Sequence
TCTCTATCACTGATAGGGA
tet operator tetO 8742..8761 Fwd
DNA Sequence
TCTCTATCACTGATAGGGA
tet operator tetO 8778..8797 Fwd
DNA Sequence
TCTCTATCACTGATAGGGA
tet operator tetO 8814..8833 Fwd
DNA Sequence
TCTCTATCACTGATAGGGA
tet operator tetO 8850..8869 Fwd
DNA Sequence
TCTCTATCACTGATAGGGA
rep_origin 1
Label Gene Product Position Strand
ori 12913..13502 Rev
DNA Sequence
TTGAGATCCTTTTTTTCTGCGCGTAATCTGCTGCTTGCAAACAAAAAAACCACCGCTACCAGCGGTGGTTTGTTTGCCGGATCAAGAGCTACCAACTCTTTTTCCGAAGGTAACTGGCTTCAGCAGAGCGCAGATACCAAATACTGTTCTTCTAGTGTAGCCGTAGTTAGGCCACCACTTCAAGAACTCTGTAGCACCGCCTACATACCTCGCTCTGCTAATCCTGTTACCAGTGGCTGCTGCCAGTGGCGATAAGTCGTGTCTTACCGGGTTGGACTCAAGACGATAGTTACCGGATAAGGCGCAGCGGTCGGGCTGAACGGGGGGTTCGTGCACACAGCCCAGCTTGGAGCGAACGACCTACACCGAACTGAGATACCTACAGCGTGAGCTATGAGAAAGCGCCACGCTTCCCGAAGGGAGAAAGGCGGACAGGTATCCGGTAAGCGGCAGGGTCGGAACAGGAGAGCGCACGAGGGAGCTTCCAGGGGGAAACGCCTGGTATCTTTATAGTCCTGTCGGGTTTCGCCACCTCTGACTTGAGCGTCGATTTTTGTGATGCTCGTCAGGGGGGCGGAGCCTATGGAAA
terminator 5
Label Gene Product Position Strand
rrnB T1 terminator Escherichia coli rrnB 98..185 Fwd
DNA Sequence
CAAATAAAACGAAAGGCTCAGTCGAAAGACTGGGCCTTTCGTTTTATCTGTTGTTTGTCGGTGAACGCTCTCCTGAGTAGGACAAAT
rrnB T2 terminator 276..304 Fwd
DNA Sequence
AGAAGGCCATCCTGACGGATGGCCTTTT
T7Te terminator 7531..7559 Rev
DNA Sequence
GGCTCACCTTCGGGTGGGCCTTTCTGCG
tonB terminator 10582..10614 Rev
DNA Sequence
AGTCAAAAGCCTCCGACCGGAGGCTTTTGACT
lambda t0 terminator 12739..12834 Rev
DNA Sequence
GACTCCTGTTGATAGATCCAGTAATGACCTCAGAACTCCATCTGGATTTGTTCAGAACGCTCGGTTGCCGCCGGGCGTTTTTTATTGGTGAGAAT