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Loc of sites 04 (320-339)

www.neb.com · info@neb.com 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 - Sac I 402
Ban II - Sac I 402
Ban II - Sac I 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

Source: http://www.bioinfo.sdu.edu.cn/czsl/pUC19_map.pdf

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