pPTR II

Autonomously replicating E. coli-Aspergillus shuttle vector with the ptrA gene conferring resistance to pyrithiamine.            

Sequence Author: TaKaRa

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HomePlasmidsBasic Cloning VectorspPTR II
NaeI (9900) NgoMIV (9898) KpnI (7045) Acc65I (7041) SmaI (7039) TspMI - XmaI (7037) SbfI (7018) HindIII (7002) lac operator AlwNI (6234) BspHI (5923) AhdI (5755) MfeI (4942) BstAPI (33) BsaAI (2736) EagI (3375) AfeI (3440) BstEII (3547) SacII (3777) BsmBI - Esp3I (3891) PflFI - Tth111I (3995) PpuMI * (4021) AgeI (4848) pPTR II 10,032 bp
NaeI  (9900)
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.
NgoMIV  (9898)
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.
KpnI  (7045)
1 site		 G G T A C C C C A T G G
Acc65I  (7041)
1 site		 G G T A C C C C A T G G
SmaI  (7039)
1 site		 C C C G G G G G G C C C
SmaI can be used at 37°C for brief incubations.
TspMI  (7037)
1 site		 C C C G G G G G G C C C
XmaI  (7037)
1 site		 C C C G G G G G G C C C
Cleavage may be enhanced when more than one copy of the XmaI recognition sequence is present.
SbfI  (7018)
1 site		 C C T G C A G G G G A C G T C C
HindIII  (7002)
1 site		 A A G C T T T T C G A A
AlwNI  (6234)
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.
BspHI  (5923)
1 site		 T C A T G A A G T A C T
AhdI  (5755)
1 site		 G A C N N N N N G T C C T G N N N N N C A G
The 1-base overhangs produced by AhdI may be hard to ligate.
Sticky ends from different AhdI sites may not be compatible.
MfeI  (4942)
1 site		 C A A T T G G T T A A C
BstAPI  (33)
1 site		 G C A N N N N N T G C C G T N N N N N A C G
Sticky ends from different BstAPI sites may not be compatible.
BsaAI  (2736)
1 site		 Y A C G T R R T G C A Y
EagI  (3375)
1 site		 C G G C C G G C C G G C
AfeI  (3440)
1 site		 A G C G C T T C G C G A
BstEII  (3547)
1 site		 G G T N A C C C C A N T G G
Sticky ends from different BstEII sites may not be compatible.
BstEII is typically used at 60°C, but is 50% active at 37°C.
SacII  (3777)
1 site		 C C G C G G G G C G C C
Efficient cleavage requires at least two copies of the SacII recognition sequence.
BsmBI  (3891)
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.
Esp3I  (3891)
1 site		 C G T C T C N G C A G A G N ( N ) 4
Sticky ends from different Esp3I sites may not be compatible.
PflFI  (3995)
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  (3995)
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.
PpuMI  (4021)
1 site		 R G G W C C Y Y C C W G G R
* Blocked by Dcm methylation.
Sticky ends from different PpuMI sites may not be compatible.
AgeI  (4848)
1 site		 A C C G G T T G G C C A
AMA1
7416 .. 2633  =  5250 bp
replication origin from Aspergillus nidulans (Aleksenko and Clutterbuck, 1996)
AMA1
7416 .. 2633  =  5250 bp
replication origin from Aspergillus nidulans (Aleksenko and Clutterbuck, 1996)
ptrA
2943 .. 4950  =  2008 bp
Product: pyrithiamine resistance gene of Aspergillus oryzae (Kubodera et al., 2000)
useful as a dominant selectable marker in Aspergillus
ptrA
2943 .. 4950  =  2008 bp
Product: pyrithiamine resistance gene of Aspergillus oryzae (Kubodera et al., 2000)
useful as a dominant selectable marker in Aspergillus
AmpR
4968 .. 5828  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
   Segment 1:  signal sequence  
   4968 .. 5036  =  69 bp
   23 amino acids  =  2.6 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
4968 .. 5828  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
   Segment 2:  
   5037 .. 5828  =  792 bp
   263 amino acids  =  28.9 kDa
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
AmpR
4968 .. 5828  =  861 bp
286 amino acids  =  31.6 kDa
2 segments
Product: β-lactamase
confers resistance to ampicillin, carbenicillin, and related antibiotics
ori
5999 .. 6587  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
ori
5999 .. 6587  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
lacZα
6985 .. 7311  =  327 bp
108 amino acids  =  12.4 kDa
Product: LacZα fragment of β-galactosidase
lacZα
6985 .. 7311  =  327 bp
108 amino acids  =  12.4 kDa
Product: LacZα fragment of β-galactosidase
lac promoter
6911 .. 6941  =  31 bp
3 segments
   Segment 1:  -35  
   6911 .. 6916  =  6 bp
promoter for the E. coli lac operon
lac promoter
6911 .. 6941  =  31 bp
3 segments
   Segment 2:  
   6917 .. 6934  =  18 bp
promoter for the E. coli lac operon
lac promoter
6911 .. 6941  =  31 bp
3 segments
   Segment 3:  -10  
   6935 .. 6941  =  7 bp
promoter for the E. coli lac operon
lac promoter
6911 .. 6941  =  31 bp
3 segments
promoter for the E. coli lac operon
lac operator
6949 .. 6965  =  17 bp
The lac repressor binds to the lac operator to inhibit transcription in E. coli. This inhibition can be relieved by adding lactose or isopropyl-β-D-thiogalactopyranoside (IPTG).
lac operator
6949 .. 6965  =  17 bp
The lac repressor binds to the lac operator to inhibit transcription in E. coli. This inhibition can be relieved by adding lactose or isopropyl-β-D-thiogalactopyranoside (IPTG).
ptrA
3187 .. 4228  =  1042 bp
327 amino acids  =  35.1 kDa
2 segments  =  984 bp
Product: pyrithiamine resistance protein (Kubodera et al., 2000)
useful as a dominant selectable marker in Aspergillus
ptrA
3187 .. 4228  =  1042 bp
327 amino acids  =  35.1 kDa
2 segments  =  984 bp
   Segment 2:  
   3187 .. 3833  =  647 bp
   215 amino acids  =  23.1 kDa
Product: pyrithiamine resistance protein (Kubodera et al., 2000)
useful as a dominant selectable marker in Aspergillus
ptrA
3187 .. 4228  =  1042 bp
327 amino acids  =  35.1 kDa
2 segments  =  984 bp
   Segment 1:  
   3892 .. 4228  =  337 bp
   112 amino acids  =  12.0 kDa
Product: pyrithiamine resistance protein (Kubodera et al., 2000)
useful as a dominant selectable marker in Aspergillus
ptrA
3187 .. 4228  =  1042 bp
327 amino acids  =  35.1 kDa
2 segments  =  984 bp
Product: pyrithiamine resistance protein (Kubodera et al., 2000)
useful as a dominant selectable marker in Aspergillus
MCS
7002 .. 7046  =  45 bp
multiple cloning site
MCS
7002 .. 7046  =  45 bp
multiple cloning site
M13 rev
6973 .. 6989  =  17 bp
common sequencing primer, one of multiple similar variants
M13 rev
6973 .. 6989  =  17 bp
common sequencing primer, one of multiple similar variants
M13 fwd
7059 .. 7075  =  17 bp
common sequencing primer, one of multiple similar variants
M13 fwd
7059 .. 7075  =  17 bp
common sequencing primer, one of multiple similar variants
ORF:  6985 .. 7311  =  327 bp
ORF:  108 amino acids  =  12.4 kDa
ORF:  2954 .. 3178  =  225 bp
ORF:  74 amino acids  =  8.4 kDa
ORF:  4968 .. 5828  =  861 bp
ORF:  286 amino acids  =  31.6 kDa
ORF:  3187 .. 3795  =  609 bp
ORF:  202 amino acids  =  21.9 kDa
ORF:  6912 .. 7268  =  357 bp
ORF:  118 amino acids  =  13.5 kDa
ORF:  3623 .. 3880  =  258 bp
ORF:  85 amino acids  =  9.7 kDa
ORF:  3884 .. 4228  =  345 bp
ORF:  114 amino acids  =  12.2 kDa
ORF:  5432 .. 5698  =  267 bp
ORF:  88 amino acids  =  9.2 kDa
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Individual Sequences & Maps

BlueScribe
pEASY-T1 (linearized)
pJET1.2
pSMART LCAmp
CAG promoter
pEASY-T1 Simple (linearized)
pJET1.2 blunt
pSMART LCAmp (linearized)
chicken beta-actin promoter
pEASY-T3 (linearized)
pJW168
pSMART LCKan
CLIP-tag
pEASY-T5 Zero (linearized)
pKF 18k-2
pSMART LCKan (linearized)
CMV promoter
pEX-1
pKF 19k-2
pSP64 Poly(A)
GAL1 promoter
pEX-A
pKOV
pSP72
GST
pEX-C-GST
pLATE11 (linearized)
pSP73
HaloTag
pEX-C-His
pLATE31 (linearized)
pSpark Done (linearized)
HSV TK promoter
pEX-K
pLATE51 (linearized)
pSpark I (linearized)
lac operator
pEX-N-GST
pLATE52 (linearized)
pSpark II (linearized)
lac promoter
pEX-N-His
pLEXX-AK
pSpark III (linearized)
lac UV5 promoter
pEX-N-His-GST
pLPS-B (linearized)
pSpark IV (linearized)
lacZ
pEX1
pMAL-c5E
pSpark V (linearized)
lambda
pEX2
pMAL-c5G
pSpot1
LITMUS28
pEX3
pMAL-c5X
pSpot2
LITMUS28i
pEZSeq-Amp
pMAL-c5X-His
pSpot3
LITMUS29
pEZSeq-Amp (linearized)
pMAL-c6T
pSpot4
LITMUS38
pEZSeq-Kan
pMAL-p5E
pSTBlue-1
LITMUS38i
pEZSeq-Kan (linearized)
pMAL-p5G
pSTV28
LITMUS39
pEZ BAC
pMAL-p5X
pSTV29
M13mp18
pEZ BAC (BamHI linearized)
pMCS5
pT7Blue
M13mp19
pEZ BAC (HpaI linearized)
pMiniT
pT7Blue-2
MBP
pF1A T7
pMiniT (linearized)
pT7Blue-3
pACYC177
pF1K T7
pMiniT 2.0
pT7CFE1-CFtag
pACYC184
pF25A ICE T7
pMiniT 2.0 (linearized)
pT7CFE1-CGFP-HA-His
pAN7-1
pF25K ICE T7
pMMB190
pT7CFE1-CGST-HA-His
pANT7_cGST
pF3A WG (BYDV)
pMMB206
pT7CFE1-CHA
pANT7_nHA
pF3K WG (BYDV)
pMMB207
pT7CFE1-CHis
pAUR316
pFC20A HaloTag T7 SP6
pMMB208
pT7CFE1-CMyc
pBC KS(+)
pFC20K HaloTag T7 SP6
pMMB66EH
pT7CFE1-NFtag
pBC KS(-)
pFC30A His6HaloTag T7
pMMB66HE
pT7CFE1-NHA
pBC SK(+)
pFC30K His6HaloTag T7
pMMB67EH
pT7CFE1-NHA-CHis
pBC SK(-)
pFC7A (HQ)
pMMB67HE
pT7CFE1-NHis
pBeloBAC11
pFC7K (HQ)
pNEB193
pT7CFE1-NHis-GST
pBluescript II KS(+)
pFN18A HaloTag T7
pNEB206A (linearized)
pT7CFE1-NHis-GST-CHA
pBluescript II KS(-)
pFN18K HaloTag T7
pOSIP-CH
pT7CFE1-NMyc
pBluescript II SK(+)
pFN19A HaloTag T7 SP6
pOSIP-CO
pTOP Blunt V2 (linearized)
pBluescript II SK(-)
pFN19K HaloTag T7 SP6
pOSIP-CT
pTrcHis A
pBluescript KS(+)
pFN29A His6HaloTag T7
pOSIP-KC
pTrcHis B
pBluescript KS(-)
pFN29K His6HaloTag T7
pOSIP-KH
pTrcHis C
pBluescript SK(+)
pFN2A (GST)
pOSIP-KL
pTrcHis2 A
pBluescript SK(-)
pFN2K (GST)
pOSIP-KO
pTrcHis2 B
pBR322
pFN6A (HQ)
pOSIP-KP
pTrcHis2 C
pBS KS(+)
pFN6K (HQ)
pOSIP-KT
pTV 118N
pBS KS(-)
pGEM-11Zf(+)
pOSIP-TH
pTWV228
pBS SK(+)
pGEM-11Zf(-)
pOSIP-TO
pTXB1
pBS SK(-)
pGEM-3Z
pOSIP-TT
pTYB21
pBS(+)
pGEM-3Zf(+)
pPCR-Script Amp SK(+)
pType IIs
pBS(-)
pGEM-3Zf(-)
pPCR-Script Cam SK(+)
pTZ19R
pBSK(+) Simple-Amp
pGEM-4Z
pPTR I
pTZ19U
pBSK(+) Simple-Kan
pGEM-5Zf(+)
pPTR II
pTZ57R
pcDNA3
pGEM-7Zf(+)
pRACE (linearized)
pUAST
pcDNA3.1(+)
pGEM-7Zf(-)
pRANGER-BTB-1
pUC118
pcDNA3.1(-)
pGEM-9Zf(-)
pRANGER-BTB-2
pUC119
pCold I
pGEM-T
pRANGER-BTB-3
pUC18
pCold II
pGEM-T (linearized)
pRANGER-BTB-5
pUC19
pCold III
pGEM-T Easy
pRHA-109
pUC19L
pCold IV
pGEM-T Easy (linearized)
pRHA-113
pUC57
pCold TF
pGGA
pRHA-67
pUC57-Kan
pCR-XL-2-TOPO
pGGAselect
pRSET A
pUC57-Simple
pCR-XL-2-TOPO (linearized)
PGK promoter
pRSET B
pUCmu
pDD
pGS-21a
pRSET C
pVLT31
pDNR-Dual
pH6HTC His6HaloTag T7
pSB1C3
pVLT33
pDNR-LIB
pH6HTN His6HaloTag T7
pSC101
pVLT35
pE-SUMOpro Amp
pHP45(omega)
pSEAP2-Basic
pYES1L
pE-SUMOpro Kan
pHP45(omega)-Km
pSF-Core
pZErO-2
pE-SUMOpro3 Amp
pHP45(omega)-Tc
pSMART BAC v2.0
SNAP-tag
pE-SUMOpro3 Kan
pHSG298
pSMART BAC v2.0 (BamHI linearized)
SP6 promoter
pE-SUMOstar Amp
pHSG299
pSMART BAC v2.0 (EcoRI linearized)
SV40 promoter
pE-SUMOstar Kan
pHSG396
pSMART BAC v2.0 (HindIII linearized)
T3 promoter
pEASY-Blunt (linearized)
pHSG398
pSMART FOS v2.0 (linearized)
T5 promoter
pEASY-Blunt E1 (linearized)
pHY300PLK
pSMART HCAmp
T7 promoter
pEASY-Blunt E2 (linearized)
pICOz
pSMART HCAmp (linearized)
tac promoter
pEASY-Blunt Simple (linearized)
pInvRecA
pSMART HCKan
trc promoter
pEASY-Blunt Zero (linearized)
pJAZZ-OC
pSMART HCKan (linearized)
phiX174
pEASY-Blunt3 (linearized)
pJAZZ-OK

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