pFB-LIC-Bse (linearized)

Linearized baculovirus vector for ligation-independent cloning (LIC), encoding an N-terminal 6xHis-TEV cassette and ampicillin resistance.

Sequence Author: Structural Genomics Consortium

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HomePlasmidsStructural Genomics VectorspFB-LIC-Bse (linearized)
RsrII (4719) BbsI (4437) PflFI - Tth111I (3908) BsmBI (3862) BstXI (3397) BspQI - SapI (3061) PspFI (2644) BseYI (2640) AlwNI (2535) NcoI (4729) ATG 6xHis End (4787) TEV site Start (0) BamHI (16) EcoRI (22) Eco53kI (30) SacI (32) SalI (35) HindIII (41) MfeI (156) BclI * (305) AvrII (320) NgoMIV (807) NaeI (809) PvuI (1687) FspI (1833) BpmI (1987) pFB-LIC-Bse 4787 bp
RsrII  (4719)
1 site		 C G G W C C G G C C W G G C
Efficient cleavage requires at least two copies of the RsrII recognition sequence.
Sticky ends from different RsrII sites may not be compatible.
For full activity, add fresh DTT.
BbsI  (4437)
1 site		 G A A G A C N N C T T C T G N N ( N ) 4
Sticky ends from different BbsI sites may not be compatible.
BbsI gradually loses activity when stored at -20°C.
PflFI  (3908)
1 site		 G A C N N N G T C C T G N N N C A G
The 1-base overhangs produced by PflFI may be hard to ligate.
Sticky ends from different PflFI sites may not be compatible.
Tth111I  (3908)
1 site		 G A C N N N G T C C T G N N N C A G
The 1-base overhangs produced by Tth111I may be hard to ligate.
Sticky ends from different Tth111I sites may not be compatible.
BsmBI  (3862)
1 site		 C G T C T C N G C A G A G N ( N ) 4
Sticky ends from different BsmBI sites may not be compatible.
BsmBI-v2 is an improved version of BsmBI.
BstXI  (3397)
1 site		 C C A N N N N N N T G G G G T N N N N N N A C C
Sticky ends from different BstXI sites may not be compatible.
BspQI  (3061)
1 site		 G C T C T T C N C G A G A A G N N N N
Sticky ends from different BspQI sites may not be compatible.
SapI  (3061)
1 site		 G C T C T T C N C G A G A A G N N N N
Sticky ends from different SapI sites may not be compatible.
SapI gradually settles in solution, so a tube of SapI should be mixed before removing an aliquot.
PspFI  (2644)
1 site		 C C C A G C G G G T C G
BseYI  (2640)
1 site		 C C C A G C G G G T C G
After cleavage, BseYI can remain bound to DNA and alter its electrophoretic mobility.
AlwNI  (2535)
1 site		 C A G N N N C T G G T C N N N G A C
Sticky ends from different AlwNI sites may not be compatible.
NcoI  (4729)
1 site		 C C A T G G G G T A C C
End  (4787)
0 sites
Start  (0)
0 sites
BamHI  (16)
1 site		 G G A T C C C C T A G G
After cleavage, BamHI-HF® (but not the original BamHI) can remain bound to DNA and alter its electrophoretic mobility.
EcoRI  (22)
1 site		 G A A T T C C T T A A G
Eco53kI  (30)
1 site		 G A G C T C C T C G A G
SacI  (32)
1 site		 G A G C T C C T C G A G
SalI  (35)
1 site		 G T C G A C C A G C T G
HindIII  (41)
1 site		 A A G C T T T T C G A A
MfeI  (156)
1 site		 C A A T T G G T T A A C
BclI  (305)
1 site		 T G A T C A A C T A G T
* Blocked by Dam methylation.
BclI is typically used at 50-55°C, but is 50% active at 37°C.
AvrII  (320)
1 site		 C C T A G G G G A T C C
NgoMIV  (807)
1 site		 G C C G G C C G G C C G
Efficient cleavage requires at least two copies of the NgoMIV recognition sequence.
NaeI  (809)
1 site		 G C C G G C C G G C C G
Efficient cleavage requires at least two copies of the NaeI recognition sequence.
PvuI  (1687)
1 site		 C G A T C G G C T A G C
FspI  (1833)
1 site		 T G C G C A A C G C G T
BpmI  (1987)
1 site		 C T G G A G ( N ) 14 N N G A C C T C ( N ) 14
Efficient cleavage requires at least two copies of the BpmI recognition sequence.
Sticky ends from different BpmI sites may not be compatible.
After cleavage, BpmI can remain bound to DNA and alter its electrophoretic mobility.
BpmI quickly loses activity at 37°C.
AmpR
1269 .. 2129  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
   Segment 1:  signal sequence  
   1269 .. 1337  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
1269 .. 2129  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
   Segment 2:  
   1338 .. 2129  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
1269 .. 2129  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
ori
2300 .. 2888  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
ori
2300 .. 2888  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
GmR
3482 .. 4015  =  534 bp
177 amino acids  =  19.4 kDa
Product: gentamycin acetyltransferase
confers resistance to gentamycin
GmR
3482 .. 4015  =  534 bp
177 amino acids  =  19.4 kDa
Product: gentamycin acetyltransferase
confers resistance to gentamycin
f1 ori
682 .. 1137  =  456 bp
f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis
f1 ori
682 .. 1137  =  456 bp
f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis
Tn7R
3191 .. 3415  =  225 bp
mini-Tn7 element (right end of the Tn7 transposon)
Tn7R
3191 .. 3415  =  225 bp
mini-Tn7 element (right end of the Tn7 transposon)
Tn7L
333 .. 498  =  166 bp
mini-Tn7 element (left end of the Tn7 transposon)
Tn7L
333 .. 498  =  166 bp
mini-Tn7 element (left end of the Tn7 transposon)
SV40 poly(A) signal
170 .. 304  =  135 bp
SV40 polyadenylation signal
SV40 poly(A) signal
170 .. 304  =  135 bp
SV40 polyadenylation signal
AmpR promoter
1164 .. 1268  =  105 bp
AmpR promoter
1164 .. 1268  =  105 bp
polyhedrin promoter
4584 .. 4675  =  92 bp
promoter for the baculovirus polyhedrin gene
polyhedrin promoter
4584 .. 4675  =  92 bp
promoter for the baculovirus polyhedrin gene
ATG
4731 .. 4733  =  3 bp
1 amino acid  =  149.2 Da
Product: start codon
ATG
4731 .. 4733  =  3 bp
1 amino acid  =  149.2 Da
Product: start codon
6xHis
4737 .. 4754  =  18 bp
6 amino acids  =  840.9 Da
Product: 6xHis affinity tag
6xHis
4737 .. 4754  =  18 bp
6 amino acids  =  840.9 Da
Product: 6xHis affinity tag
TEV site
4779 .. 4799  =  21 bp
7 amino acids  =  900.0 Da
Product: tobacco etch virus (TEV) protease recognition and cleavage site
TEV site
4779 .. 4799  =  21 bp
7 amino acids  =  900.0 Da
Product: tobacco etch virus (TEV) protease recognition and cleavage site
Pc promoter
4204 .. 4232  =  29 bp
3 segments
   Segment 3:  -10  
   4204 .. 4209  =  6 bp
class 1 integron promoter
Pc promoter
4204 .. 4232  =  29 bp
3 segments
   Segment 2:  
   4210 .. 4226  =  17 bp
class 1 integron promoter
Pc promoter
4204 .. 4232  =  29 bp
3 segments
   Segment 1:  -35  
   4227 .. 4232  =  6 bp
class 1 integron promoter
Pc promoter
4204 .. 4232  =  29 bp
3 segments
class 1 integron promoter
ORF:  3325 .. 3627  =  303 bp
ORF:  100 amino acids  =  11.8 kDa
ORF:  4117 .. 4596  =  480 bp
ORF:  159 amino acids  =  18.5 kDa
ORF:  3620 .. 3952  =  333 bp
ORF:  110 amino acids  =  11.9 kDa
ORF:  4226 .. 4468  =  243 bp
ORF:  80 amino acids  =  9.1 kDa
ORF:  1269 .. 2129  =  861 bp
ORF:  286 amino acids  =  31.6 kDa
ORF:  1733 .. 1999  =  267 bp
ORF:  88 amino acids  =  9.2 kDa
ORF:  3482 .. 4015  =  534 bp
ORF:  177 amino acids  =  19.4 kDa
ORF:  4207 .. 4515  =  309 bp
ORF:  102 amino acids  =  11.4 kDa
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Individual Sequences & Maps

p11
pMCSG19B (linearized)
pMCSG41
pMCSG68 (linearized)
p15Tv-L
pMCSG19C
pMCSG41 (linearized)
pMCSG69
p283
pMCSG19C (linearized)
pMCSG42
pMCSG69 (linearized)
pBEN1-SGC
pMCSG20
pMCSG42 (linearized)
pMCSG7
pBEN1-SGC (linearized)
pMCSG20 (linearized)
pMCSG43
pMCSG7 (linearized)
pDB-HisGST
pMCSG21
pMCSG43 (linearized)
pMCSG70
pDB.GST
pMCSG21 (linearized)
pMCSG46
pMCSG70 (linearized)
pDB.His.MBP
pMCSG22
pMCSG46 (linearized)
pMCSG71
pDB.His.NusA
pMCSG22 (linearized)
pMCSG48
pMCSG71 (linearized)
pDB.His.TRX
pMCSG23
pMCSG48 (linearized)
pMCSG73
pDB.TRX
pMCSG23 (linearized)
pMCSG49
pMCSG73 (linearized)
pEU-GST-FV
pMCSG24
pMCSG49 (linearized)
pMCSG75
pEU-His-FV
pMCSG24 (linearized)
pMCSG50
pMCSG75 (linearized)
pEU-HSBC
pMCSG25
pMCSG50 (linearized)
pMCSG76
pFB-LIC-Bse
pMCSG25 (linearized)
pMCSG51
pMCSG76 (linearized)
pFB-LIC-Bse (linearized)
pMCSG26
pMCSG51 (linearized)
pMCSG77
pH3c-LIC
pMCSG26 (linearized)
pMCSG52
pMCSG77 (linearized)
pH3c-LIC (linearized)
pMCSG27
pMCSG52 (linearized)
pMCSG79
pHM3C-LIC
pMCSG27 (linearized)
pMCSG53
pMCSG79 (linearized)
pHM3C-LIC (linearized)
pMCSG28
pMCSG53 (linearized)
pMCSG8
pLIC-CH
pMCSG28 (linearized)
pMCSG54
pMCSG8 (linearized)
pLIC-CH (linearized)
pMCSG29
pMCSG54 (linearized)
pMCSG81
pLIC-EGFP
pMCSG29 (linearized)
pMCSG55
pMCSG81 (linearized)
pLIC-EGFP (linearized)
pMCSG30
pMCSG55 (linearized)
pMCSG9
pLIC-SGC1
pMCSG30 (linearized)
pMCSG56
pMCSG9 (linearized)
pLIC-SGC1 (linearized)
pMCSG31
pMCSG56 (linearized)
pNH-TrxT
pMCentr1
pMCSG31 (linearized)
pMCSG57
pNH-TrxT (linearized)
pMCentr1 (linearized)
pMCSG32
pMCSG57 (linearized)
pNIC-CH
pMCentr2
pMCSG32 (linearized)
pMCSG58
pNIC-CH (linearized)
pMCentr2 (linearized)
pMCSG33
pMCSG58 (linearized)
pNIC-CTHF
pMCentr3
pMCSG33 (linearized)
pMCSG59
pNIC-CTHF (linearized)
pMCentr3 (linearized)
pMCSG34
pMCSG59 (linearized)
pNIC-Zb
pMCSG10
pMCSG34 (linearized)
pMCSG60
pNIC-Zb (linearized)
pMCSG10 (linearized)
pMCSG34B
pMCSG60 (linearized)
pNIC28-Bsa4
pMCSG11
pMCSG34B (linearized)
pMCSG61
pNIC28-Bsa4 (linearized)
pMCSG11 (linearized)
pMCSG35
pMCSG61 (linearized)
pNYCOMPS-LIC-FH10T+ (N term) - linearized
pMCSG12
pMCSG35 (linearized)
pMCSG62
pNYCOMPS-LIC-FH10T+ (N term)
pMCSG12 (linearized)
pMCSG35B
pMCSG62 (linearized)
pSpeedET
pMCSG13
pMCSG35B (linearized)
pMCSG63
pVP13
pMCSG13 (linearized)
pMCSG37
pMCSG63 (linearized)
pVP16
pMCSG14
pMCSG37 (linearized)
pMCSG64
pVP33A
pMCSG14 (linearized)
pMCSG38
pMCSG64 (linearized)
pVP33K
pMCSG17
pMCSG38 (linearized)
pMCSG65
pVP56A
pMCSG17 (linearized)
pMCSG38C
pMCSG65 (linearized)
pVP56K
pMCSG18
pMCSG38C (linearized)
pMCSG66
pVP65K
pMCSG18 (linearized)
pMCSG39
pMCSG66 (linearized)
pVP68K
pMCSG19
pMCSG39 (linearized)
pMCSG67
pVP81K
pMCSG19 (linearized)
pMCSG40
pMCSG67 (linearized)
pVP91A
pMCSG19B
pMCSG40 (linearized)
pMCSG68

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