pOSIP-TH

Prokaryotic one-step cloning and chromosomal integration vector encoding a tetracycline resistance marker and the HK022 integrase.
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HomePlasmidsBasic Cloning VectorspOSIP-TH
EcoRI (1) rrnB T1 terminator rrnB T1 terminator rrnB T1 terminator rrnB T1 terminator PflFI - Tth111I (6096) BseRI (5748) BmtI (5624) NheI (5620) NdeI (5351) SalI (4853) StuI (4714) ScaI (4320) BglI (4277) PmlI (4043) SspI (3991) EcoRV (3601) BspDI - ClaI (3421) Acc65I (13) KpnI (17) BamHI (22) BssHII (70) SrfI (225) BsaI (363) BglII (469) BciVI (923) SpeI (1126) PstI (1144) SphI (1150) PaeR7I - XhoI (1152) BstBI (1234) PacI (1340) PspOMI (1349) ApaI (1353) ZraI (1379) AatII (1381) AvrII (1418) BsmI (1718) PshAI (2090) KflI - PpuMI (2350) NruI (2828) BtsI - BtsαI (3076) DraIII (3415) pOSIP-TH 6925 bp
EcoRI  (1)
1 site		 G A A T T C C T T A A G
PflFI  (6096)
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  (6096)
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.
BseRI  (5748)
1 site		 G A G G A G ( N ) 8 N N C T C C T C ( N ) 8
Sticky ends from different BseRI sites may not be compatible.
BseRI quickly loses activity at 37°C.
Prolonged incubation with BseRI may lead to degradation of the DNA.
BmtI  (5624)
1 site		 G C T A G C C G A T C G
NheI  (5620)
1 site		 G C T A G C C G A T C G
NdeI  (5351)
1 site		 C A T A T G G T A T A C
Prolonged incubation with NdeI may lead to removal of additional nucleotides.
SalI  (4853)
1 site		 G T C G A C C A G C T G
StuI  (4714)
1 site		 A G G C C T T C C G G A
ScaI  (4320)
1 site		 A G T A C T T C A T G A
BglI  (4277)
1 site		 G C C N N N N N G G C C G G N N N N N C C G
Sticky ends from different BglI sites may not be compatible.
PmlI  (4043)
1 site		 C A C G T G G T G C A C
SspI  (3991)
1 site		 A A T A T T T T A T A A
EcoRV  (3601)
1 site		 G A T A T C C T A T A G
EcoRV is reportedly more prone than its isoschizomer Eco32I to delete a base after cleavage.
BspDI  (3421)
1 site		 A T C G A T T A G C T A
ClaI  (3421)
1 site		 A T C G A T T A G C T A
Acc65I  (13)
1 site		 G G T A C C C C A T G G
KpnI  (17)
1 site		 G G T A C C C C A T G G
BamHI  (22)
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.
BssHII  (70)
1 site		 G C G C G C C G C G C G
BssHII is typically used at 50°C, but is 75% active at 37°C.
SrfI  (225)
1 site		 G C C C G G G C C G G G C C C G
BsaI  (363)
1 site		 G G T C T C N C C A G A G N ( N ) 4
Sticky ends from different BsaI sites may not be compatible.
BsaI can be used between 37°C and 50°C.
BglII  (469)
1 site		 A G A T C T T C T A G A
BciVI  (923)
1 site		 G T A T C C ( N ) 5 N C A T A G G ( N ) 5
The 1-base overhangs produced by BciVI may be hard to ligate.
Sticky ends from different BciVI sites may not be compatible.
SpeI  (1126)
1 site		 A C T A G T T G A T C A
PstI  (1144)
1 site		 C T G C A G G A C G T C
SphI  (1150)
1 site		 G C A T G C C G T A C G
PaeR7I  (1152)
1 site		 C T C G A G G A G C T C
PaeR7I does not recognize the sequence CTCTCGAG.
XhoI  (1152)
1 site		 C T C G A G G A G C T C
BstBI  (1234)
1 site		 T T C G A A A A G C T T
PacI  (1340)
1 site		 T T A A T T A A A A T T A A T T
PspOMI  (1349)
1 site		 G G G C C C C C C G G G
ApaI  (1353)
1 site		 G G G C C C C C C G G G
ApaI can be used between 25°C and 37°C.
ZraI  (1379)
1 site		 G A C G T C C T G C A G
AatII  (1381)
1 site		 G A C G T C C T G C A G
AvrII  (1418)
1 site		 C C T A G G G G A T C C
BsmI  (1718)
1 site		 G A A T G C N C T T A C G N
Sticky ends from different BsmI sites may not be compatible.
PshAI  (2090)
1 site		 G A C N N N N G T C C T G N N N N C A G
PshAI quickly loses activity at 37°C, but can be used at 25°C for long incubations.
KflI  (2350)
1 site		 G G G W C C C C C C W G G G
Sticky ends from different KflI sites may not be compatible.
PpuMI  (2350)
1 site		 R G G W C C Y Y C C W G G R
Sticky ends from different PpuMI sites may not be compatible.
NruI  (2828)
1 site		 T C G C G A A G C G C T
BtsI  (3076)
1 site		 G C A G T G N N C G T C A C
BtsαI  (3076)
1 site		 G C A G T G N N C G T C A C
Sticky ends from different BtsαI sites may not be compatible.
DraIII  (3415)
1 site		 C A C N N N G T G G T G N N N C A C
Sticky ends from different DraIII sites may not be compatible.
TcR
3488 .. 4693  =  1206 bp
401 amino acids  =  43.4 kDa
Product: tetracycline efflux protein
confers resistance to tetracycline
TcR
3488 .. 4693  =  1206 bp
401 amino acids  =  43.4 kDa
Product: tetracycline efflux protein
confers resistance to tetracycline
HK022 integrase
2270 .. 3343  =  1074 bp
357 amino acids  =  40.4 kDa
Product: integrase from phage HK022
HK022 integrase
2270 .. 3343  =  1074 bp
357 amino acids  =  40.4 kDa
Product: integrase from phage HK022
λ repressor (ts)
1424 .. 2137  =  714 bp
237 amino acids  =  26.2 kDa
Product: temperature-sensitive variant of the phage λ repressor
thermosensitivity is conferred by the A67T mutation
λ repressor (ts)
1424 .. 2137  =  714 bp
237 amino acids  =  26.2 kDa
Product: temperature-sensitive variant of the phage λ repressor
thermosensitivity is conferred by the A67T mutation
ori
477 .. 1065  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
ori
477 .. 1065  =  589 bp
high-copy-number ColE1/pMB1/pBR322/pUC origin of replication
R6K γ ori
4903 .. 5291  =  389 bp
γ replication origin from E. coli plasmid R6K; requires the R6K initiator protein pi for replication
R6K γ ori
4903 .. 5291  =  389 bp
γ replication origin from E. coli plasmid R6K; requires the R6K initiator protein pi for replication
ccdB
142 .. 447  =  306 bp
101 amino acids  =  11.7 kDa
Product: CcdB, a bacterial toxin that poisons DNA gyrase
Plasmids containing the ccdB gene cannot be propagated in standard E. coli strains.
ccdB
142 .. 447  =  306 bp
101 amino acids  =  11.7 kDa
Product: CcdB, a bacterial toxin that poisons DNA gyrase
Plasmids containing the ccdB gene cannot be propagated in standard E. coli strains.
λ tL3 terminator
5640 .. 5886  =  247 bp
transcription terminator tL3 from phage λ
λ tL3 terminator
5640 .. 5886  =  247 bp
transcription terminator tL3 from phage λ
HK022 attP
5379 .. 5605  =  227 bp
attachment site of phage HK022
HK022 attP
5379 .. 5605  =  227 bp
attachment site of phage HK022
lambda t0 terminator
4752 .. 4846  =  95 bp
transcription terminator from phage lambda
lambda t0 terminator
4752 .. 4846  =  95 bp
transcription terminator from phage lambda
rrnB T1 terminator
6206 .. 6292  =  87 bp
transcription terminator T1 from the E. coli rrnB gene
rrnB T1 terminator
6206 .. 6292  =  87 bp
transcription terminator T1 from the E. coli rrnB gene
rrnB T1 terminator
6387 .. 6473  =  87 bp
transcription terminator T1 from the E. coli rrnB gene
rrnB T1 terminator
6387 .. 6473  =  87 bp
transcription terminator T1 from the E. coli rrnB gene
rrnB T1 terminator
6568 .. 6654  =  87 bp
transcription terminator T1 from the E. coli rrnB gene
rrnB T1 terminator
6568 .. 6654  =  87 bp
transcription terminator T1 from the E. coli rrnB gene
rrnB T1 terminator
6749 .. 6835  =  87 bp
transcription terminator T1 from the E. coli rrnB gene
rrnB T1 terminator
6749 .. 6835  =  87 bp
transcription terminator T1 from the E. coli rrnB gene
MCS 1
1 .. 75  =  75 bp
multiple cloning site, part 1
MCS 1
1 .. 75  =  75 bp
multiple cloning site, part 1
tetR/tetA promoters
3426 .. 3480  =  55 bp
overlapping promoters for bacterial tetR and tetA
tetR/tetA promoters
3426 .. 3480  =  55 bp
overlapping promoters for bacterial tetR and tetA
FRT
5302 .. 5349  =  48 bp
FLP-mediated recombination occurs in the 8-bp core sequence TCTAGAAA (Turan and Bode, 2011).
FRT
5302 .. 5349  =  48 bp
FLP-mediated recombination occurs in the 8-bp core sequence TCTAGAAA (Turan and Bode, 2011).
FRT (minimal)
1384 .. 1417  =  34 bp
supports FLP-mediated excision but not integration (Turan and Bode, 2011)
FRT (minimal)
1384 .. 1417  =  34 bp
supports FLP-mediated excision but not integration (Turan and Bode, 2011)
MCS 2
1126 .. 1157  =  32 bp
multiple cloning site, part 2
MCS 2
1126 .. 1157  =  32 bp
multiple cloning site, part 2
tonB terminator
1276 .. 1307  =  32 bp
bidirectional E. coli tonB-P14 transcription terminator
tonB terminator
1276 .. 1307  =  32 bp
bidirectional E. coli tonB-P14 transcription terminator
rrnB T2 terminator
1200 .. 1227  =  28 bp
transcription terminator T2 from the E. coli rrnB gene
rrnB T2 terminator
1200 .. 1227  =  28 bp
transcription terminator T2 from the E. coli rrnB gene
tet operator
3461 .. 3479  =  19 bp
bacterial operator O2 for the tetR and tetA genes
tet operator
3461 .. 3479  =  19 bp
bacterial operator O2 for the tetR and tetA genes
ORF:  142 .. 447  =  306 bp
ORF:  101 amino acids  =  11.7 kDa
ORF:  2270 .. 3343  =  1074 bp
ORF:  357 amino acids  =  40.4 kDa
ORF:  3488 .. 4693  =  1206 bp
ORF:  401 amino acids  =  43.4 kDa
ORF:  6122 .. 6361  =  240 bp
ORF:  79 amino acids  =  9.2 kDa
ORF:  3150 .. 3407  =  258 bp
ORF:  85 amino acids  =  9.5 kDa
ORF:  1424 .. 2137  =  714 bp
ORF:  237 amino acids  =  26.2 kDa
ORF:  2659 .. 2940  =  282 bp
ORF:  93 amino acids  =  9.8 kDa
ORF:  3862 .. 4122  =  261 bp
ORF:  86 amino acids  =  9.3 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

Plasmid Sets

Basic Cloning Vectors
I.M.A.G.E. Consortium Plasmids
pET & Duet Vectors (Novagen)
TA and GC Cloning Vectors
Coronavirus Resources
Insect Cell Vectors
pGEX Vectors (GE Healthcare)
TOPO Cloning Vectors
CRISPR Plasmids
Luciferase Vectors
Plant Vectors
Viral Expression & Packaging Vectors
Fluorescent Protein Genes & Plasmids
Lucigen Vectors
Qiagen Vectors
Yeast Plasmids
Gateway® Cloning Vectors
Mammalian Expression Vectors
Structural Genomics Vectors

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