Home  Contacts  Order  Catalog  Support  Search  Alphabetical Index  Numerical Index  Restriction Enzymes  Modifying Enzymes  PCR, qPCR, RT-PCR & dNTPs  Molecular Cloning  Nucleic Acid Purification  Molecular Labeling & Detection  In vitro Transcription  Electrophoresis Products  Nucleotides  Transfection Reagents  Reagents S U P P O R T

pBluescript II KS(+/-), pBluescript II SK(+/-): description & restriction map

Related Documents: pBluescript II KS(-) GenBank/EMBL accession number X52329. Sequence pBluescript II KS(+) GenBank/EMBL accession number X52327. Restriction sites Sequence pBluescript II SK(-) GenBank/EMBL accession number X52330. Sequence pBluescript II SK(+) GenBank/EMBL accession number X52328. Restriction sites Sequence Not available from Fermentas

Additional Information:

• CAP protein binding site - 938-901 (compl. strand);

• mRNA (LacZ) starts at nt position 854 (compl. strand);
• lac repressor binding site - 854-834 (compl. strand);
• f1 packaging signal [pBluescript II SK/KS(+)] - 366-457;
• f1 packaging signal [pBluescript II SK/KS(-)] - 5-96.

All pBluescript II phagemids are 2961 bp in length. The structure of these phagemids is very similar to that of pTZ19R/U (see pTZ19R/U page for information) except for the MCSs and the promoters flanking the MCS. pBluescript II phagemids are designed for DNA cloning, dideoxy DNA sequencing, in vitro mutagenesis and in vitro transcription in a single system. The pBluescript II SK and KS vector series represent two orientations of the MCS within the lacZ gene encoding the N-terminal fragment of beta-galactosidase (KS represents the orientation of the MCS in which lacZ transcription proceeds from KpnI to SacI, while SK - from SacI to KpnI). (+) and (-) symbols on pBluescript II phagemids indicate the orientation of the cloned phage f1 intergenic (IG) region carrying the sequences required in cis for initiation and termination of phage f1 DNA synthesis and for packaging of DNA into bacteriophage particles. Synthesis of single-stranded DNA requires the phage-encoded gene II, X and V proteins and is initiated at ori (+). It proceeds in the direction indicated. The conversion of single-stranded DNA strands to double strands does not require any of the phage genes to function. DNA synthesis is initiated by a 30-nucleotide RNA primer synthesized by host's RNA polymerase and starting at ori (-). pBluescript II phagemids contain: (1) f1 (IG) - the intergenic region of phage f1; (2) rep (pMB1) - the pMB1 replicon responsible for the replication of phagemid. The indicated region is sufficient to promote replication. DNA replication initiates at position 1213 (+/- 1) and proceeds in the direction indicated; (3) bla (ApR) - gene, coding for beta-lactamase that confers resistance to ampicillin. Nucleotides 2833-2765 (complementary strand) code for a signal peptide; (4) lacZ - 5'-terminal part of lacZ gene encoding the N-terminal fragment of beta-galactosidase. This fragment allows blue/white screening of recombinant phagemids. The LacZ polypeptide corresponding to wt beta-galactosidase and essential for blue/white screening ends at nt position 460 (complementary strand). Other codons in the same reading frame come from f1 DNA.

The maps show enzymes that cut pBluescript II SK/KS(+) and pBluescript II SK/KS(-) DNA once. Enzymes produced by Fermentas are shown in blue. The coordinates refer to the position of first nucleotide in each recognition sequence. The exact position of genetic elements is shown on the map (termination codons included).

References

1. Alting-Mees, M.A. and Short, J.M., pBluescript II: gene mapping vectors, Nucleic Acids Res., 17, 9494, 1989.

2. Alting-Mees, M.A., Sorge, J.A. and Short, J.M., pBluescriptII: multifunctional cloning and mapping vectors, Meth. Enzymol., 216, 483-495, 1992.

3. Short, J.M., Fernandez, J.M., Sorge, J.A. and Huse, W.D., Lambda ZAP: a bacteriophage lambda expression vector with in vivo excision properties, Nucleic Acids Res., 16, 7583-7600, 1988.

Enzymes which cut pBluescript II KS(+) DNA once:
Acc65I 755, AdeI 222, AflIII 1153, AloI 169, ApaI 749, BamHI 689, BcgI 2562, BcuI 683, BsaAI 225, Bsp120I 749, BstXI 658, Bsu15I 725, BtgI 661, CaiI 1564, Cfr9I 695, Cfr42I 661, Eam1105I 2041, Ecl136II 653, Eco31I 2113, Eco32I 713, Eco52I 670, EcoO109I 748, EcoRI 707, FaqI 457, GsuI 2131, Hin1I 2582, HincII 734, HindIII 719, KpnI 755, NotI 669, OliI 659, PdiI 328, PdmI 2641, PscI 1153, PsiI 97, PstI 701, SacI 653, SalI 734, SapI 1030, ScaI 2524, SmaI 695, TatI 2524, XbaI 677, XceI 1153, XhoI 740, XmiI 734.

Enzymes which cut pBluescript II SK(-) DNA once:
Acc65I 653, AdeI 232, AflIII 1153, AloI 281, ApaI 659, BamHI 719, BcgI 2562, BcuI 725, BsaAI 232, BseYI 1457, Bsp120I 659, BstXI 744, Bsu15I 683, BtgI 747, CaiI 1564, Cfr9I 713, Cfr42I 747, Eam1105I 2041, Ecl136II 755, Eco31I 2113, Eco32I 695, Eco52I 738, EcoO109I 659, EcoRI 701, Hin1I 2582, HincII 674, HindIII 689, KpnI 653, NotI 737, OliI 745, PdiI 129, PdmI 2641, PscI 1153, PsiI 360, PstI 707, SacI 755, SalI 674, SapI 1030, ScaI 2524, SmaI 713, TatI 2524, XbaI 731, XceI 1153, XhoI 668, XmiI 674.

Multiple Cloning Sites

pBluescript II KS(-), pBluescript II KS(+)

pBluescript II SK(-), pBluescript II SK(+)

catalog@fermentas.com

Updated lapkričio 08, 2006 14:22