www.neb.com · [email protected] TECHNICAL SUPPORT
Enzyme # Locations Enzyme # Locations
pUC19 is a small, high-copy number E. coli plasmid cloning
Enzymes with unique restriction sites are shown in bold type
vector containing portions of pBR322 and M13mp19 (1). It
and enzymes with two restriction sites are shown in regular
See page 141 for ordering information.
contains the pMB1 origin of replication from pBR322 but lacks
type. The accompanying table shows restriction sites of those
the rop gene and carries a point mutation in the RNAII transcript
enzymes that cut a moderate number of times. Restriction site
Feature Coordinates Source
(G 2975 in pBR322 to A 1308 in pUC19; 2). These changes
coordinates refer to the position of the 5´-most base on the top
together result in a temperature-dependent copy number of
strand in each recognition sequence.
about 75 per cell at 37°C and >200 per cell at 42°C (2, 3). The
Open reading frame (ORF) coordinates are in the form
multiple cloning site (MCS) is in frame with the lacZα gene,
"translational start – translational stop”; numbers refer to
allowing screening for insertions using α-complementation.
positions on the top (clockwise) strand, regardless of the
pUC18 is identical to pUC19 except that the MCS region
direction of transcription and include the start and stop codons.
Origin of replication coordinates include the region from the
References
pNEB193 (NEB #N3051S) is also identical to pUC19 except
-35 promoter sequence of the RNAII transcript to the RNA/DNA
1. Yanisch-Perron, C., Vieira, J. and Messing, J.
for the addition of several restriction endonuclease sites to the
switch point. bla (ApR) gene coordinates include the signal
2. Lin-Chao, S., Chen, W.-T. and Wong, T.-T.
(1992) Mol. Microbiol. 6, 3385–3393
3. Miki, T. et al. (1987) Protein Eng. 1, 327–332. BstAP I 179 BstAP I 179 BstAP I 179 EcoO109 I Kas I - Nar I - SfKas I - Nar I - Sf EcoO109 I 2674 Kas I - Nar I - Sfo I 235 Aat II - Zra I 2617 Aat II - Zra I 2617 Aat II - Zra I 2617 Ssp I 2501 Ssp I Ssp I 2501 lacZ Apo I - EcoR I 396 Xmn I 2294 Apo I - EcoR I Apo I - EcoR I 396 lacZ lacZ Ban II - SacI 402 Ban II - SacI 402 Ban II - SacI 402 Acc65 I - Kpn I 408 Bcg I 2215 Acc65 I - Kpn I 408 Acc65 I - Kpn I 408 Bcg I 2215Bcg I 2215 MCS Ava I - BsoB I - a I - BsoB I - Ava I - BsoB I - Sca I 2177 Sma I - Xma I 412 Sca I 2177 Sca I 2177 Sma I - Xma I Sma I - Xma BamH I 417 BamH I 417 BamH I 417 Acc I - Hinc II - Sal I Xba Acc I - Hinc II - Sal I BspM I - BfuA I 433 Acc I - Hinc II - Sal BspM I - BfuA I 433 pUC19 pUC19 BspM I - BfuA I 433 Hind III 447 Hind III 447 Hind III 447 BsaX I 659 BsaX I 659 Bpm I 1784 BsaX I 659 BsrF I 1779 Bsa I 1766 Afl III - Pci I 806 Afl III - Pci I 806 Afl III - Pci I 806 Ahd I 1694 Ahd I 1694 BseY I 1110 BseY I 1110 AlwN I 1217 AlwN I 1217 BseY I 1110 AlwN I 1217 pUC19 MCS pUC19 MCS SacI SmaI XbaI SbfI pUC19 MCS SacI SmaI XbaI SbfI EcoRI KpnI BamHI SalI PstI SphI HindIII EcoRI KpnI BamHI SalI PstI SphI HindIII agtgAATTCGAGCTCGGTACCCGGGGATCCTCTAGAGTCGACCTGCAGGCATGCAAGCTTGGcgtaatcatggtcat SacI SmaI XbaI SbfI agtgAATTCGAGCTCGGTACCCGGGGATCCTCTAGAGTCGACCTGCAGGCATGCAAGCTTGGcgtaatcatggtcat EcoRI KpnI BamHI SalI PstI SphI HindIII 4OO 41O 42O 43O 44O 45O 46O agtgAATTCGAGCTCGGTACCCGGGGATCCTCTAGAGTCGACCTGCAGGCATGCAAGCTTGGcgtaatcatggtcat 4OO 41O 42O 43O 44O 45O 46O .S N S S P V R P D E L T S R C A H L S P T I M T M 4OO 41O 42O 43O 44O 45O 46O .S N S S P V R P D E L T S R C A H L S P T I M T M .S N S S P V R P D E L T S R C A H L S P T I M T M lacZα translational start
lacZα translational start
lacZα translational start
pNEB193 MCS pNEB193 MCS SacI SmaI BssHII PacI SbfI SacI SmaI BssHII PacI SbfI EcoRI KpnI AscI BamHI XbaI SalI PmeI PstI SphI HindIII pNEB193 MCS EcoRI KpnI AscI BamHI XbaI SalI PmeI PstI SphI HindIII agtgAATTCGAGCTCGGTACCCGGGGGCGCGCCGGATCCTTAATTAAGTCTAGAGTCGACTGTTTAAACCTGCAGGCATGCAAGCTTGGcgtaatcatggtcat agtgAATTCGAGCTCGGTACCCGGGGGCGCGCCGGATCCTTAATTAAGTCTAGAGTCGACTGTTTAAACCTGCAGGCATGCAAGCTTGGcgtaatcatggtcat . . . . . . . . . . SacI SmaI BssHII PacI SbfI . . . . . . . . . . 4OO 41O 42O 43O 44O 45O 46O 47O 48O 49O EcoRI KpnI AscI BamHI XbaI SalI PmeI PstI SphI HindIII 4OO 41O 42O 43O 44O 45O 46O 47O 48O 49O agtgAATTCGAGCTCGGTACCCGGGGGCGCGCCGGATCCTTAATTAAGTCTAGAGTCGACTGTTTAAACCTGCAGGCATGCAAGCTTGGcgtaatcatggtcat .S N S S P V R P R A P D K I L D L T S Q K F R C A H L S P T I M T M . . . . . . . . . . .S N S S P V R P R A P D K I L D L T S Q K F R C A H L S P T I M T M 4OO 41O 42O 43O 44O 45O 46O 47O 48O 49O lacZα translational start
.S N S S P V R P R A P D K I L D L T S Q K F R C A H L S P T I M T M lacZα translational start
lacZα translational start
Umberto Veronesi Umberto Veronesi is a surgeon who has devoted the greater part of his professional life to exploring the paths of research with the aim of improving the treatment and the quality of life for cancer patients. It was he who, on the basis of controlled clinical trials, first contributed at the National Cancer Institute in Milan to the development of the conservative treatment
JORNADAS DE ONCOLOGÍA Y ATENCIÓN PRIMARIA CONTROL DE SÍNTOMAS GRUPO DE PALIATIVOS DE LA SAMFYC PRINCIPIOS GENERALES INTRODUCCIÓN En nuestro medio tres de cada mil personas fallecen cada año por cáncer. A pesar de los avances en prevención, diagnóstico y tratamiento, la enfermedad oncológica sólo es curable en menos de un 50% de los casos; además de ser u