pcdna™3.1/hygro(+) - tsv-co.com plasmid/pcdna3.1... · pcdna3.1 hygro(+).dna (circular / 5597 bp)...
TRANSCRIPT
Unique Cutters Bo ldSequence: pcDNA3.1 Hygro(+).dna (Circular / 5597 bp)Enzymes: Unique Cutters + BstXI (49 of 646 total)Features: 14 visible, 14 total
Printed from SnapGene® Viewer: Tuesday, May 8, 2012 2:32:05 PM Page 1
Mammalian expression vector with the CMV promoter. The MCS is in the forward (+) orientation.
SgrDI(5595)
SspI(5478)
FspI(4896)
A hdI(4674)
P ciI(3784)
BspQI - SapI(3668)
BstZ17I(3405)BsmI(3353)
P f oI(3208)
SacII(2898)
BglI I (12)
Mf eI (161)Bpu10I (180)
NruI (208)MluI (228)
SnaBI (590)
NheI (895)Bmt I (899)
A f lI I (908)HindII I (911)
A cc65I (917)KpnI (921)BamHI (929)BstXI (948)EcoRV (964)BstXI (974)Not I (979)PaeR7I - P spXI - TliI - XhoI (985)XbaI (991)EcoO109I - P spOMI (997)A paI (1001)
BbsI (1217)
SexA I* (1821)
BseRI (2050)StuI (2053)
A vrII (2054)TspMI - XmaI (2075)
SmaI (2077)P shA I (2145)
BsmBI (2186)
Bf uA I - BspMI (2431)A siSI (2482)
RsrI I (2526)
pcDNA ™3.1/ Hygro(+)5597 bp
pcDNA3.1 Hygro(+).dna (Circular / 5597 bp)
Printed from SnapGene® Viewer: Tuesday, May 8, 2012 2:32:06 PM Page 2
BglI I
5 ʹ
3 ʹ85
G A C G G A T C G G G A G A T C T C C C G A T C C C C T A T G G T G C A C T C T C A G T A C A A T C T G C T C T G A T G C C G C A T A G T T A A G C C A G T A T C T G C T
C T G C C T A G C C C T C T A G A G G G C T A G G G G A T A C C A C G T G A G A G T C A T G T T A G A C G A G A C T A C G G C G T A T C A A T T C G G T C A T A G A C G A
Mf eI
170C C C T G C T T G T G T G T T G G A G G T C G C T G A G T A G T G C G C G A G C A A A A T T T A A G C T A C A A C A A G G C A A G G C T T G A C C G A C A A T T G C A T G
G G G A C G A A C A C A C A A C C T C C A G C G A C T C A T C A C G C G C T C G T T T T A A A T T C G A T G T T G T T C C G T T C C G A A C T G G C T G T T A A C G T A C
MluINruIBpu10I
255
C MV enhancer
A A G A A T C T G C T T A G G G T T A G G C G T T T T G C G C T G C T T C G C G A T G T A C G G G C C A G A T A T A C G C G T T G A C A T T G A T T A T T G A C T A G T T
T T C T T A G A C G A A T C C C A A T C C G C A A A A C G C G A C G A A G C G C T A C A T G C C C G G T C T A T A T G C G C A A C T G T A A C T A A T A A C T G A T C A A
340
C MV enhancer
A T T A A T A G T A A T C A A T T A C G G G G T C A T T A G T T C A T A G C C C A T A T A T G G A G T T C C G C G T T A C A T A A C T T A C G G T A A A T G G C C C G C C
T A A T T A T C A T T A G T T A A T G C C C C A G T A A T C A A G T A T C G G G T A T A T A C C T C A A G G C G C A A T G T A T T G A A T G C C A T T T A C C G G G C G G
425
C MV enhancer
T G G C T G A C C G C C C A A C G A C C C C C G C C C A T T G A C G T C A A T A A T G A C G T A T G T T C C C A T A G T A A C G C C A A T A G G G A C T T T C C A T T G A
A C C G A C T G G C G G G T T G C T G G G G G C G G G T A A C T G C A G T T A T T A C T G C A T A C A A G G G T A T C A T T G C G G T T A T C C C T G A A A G G T A A C T
510
C MV enhancer
C G T C A A T G G G T G G A G T A T T T A C G G T A A A C T G C C C A C T T G G C A G T A C A T C A A G T G T A T C A T A T G C C A A G T A C G C C C C C T A T T G A C G
G C A G T T A C C C A C C T C A T A A A T G C C A T T T G A C G G G T G A A C C G T C A T G T A G T T C A C A T A G T A T A C G G T T C A T G C G G G G G A T A A C T G C
SnaBI
595
C MV enhancer
T C A A T G A C G G T A A A T G G C C C G C C T G G C A T T A T G C C C A G T A C A T G A C C T T A T G G G A C T T T C C T A C T T G G C A G T A C A T C T A C G T A T T
A G T T A C T G C C A T T T A C C G G G C G G A C C G T A A T A C G G G T C A T G T A C T G G A A T A C C C T G A A A G G A T G A A C C G T C A T G T A G A T G C A T A A
680
C MV enhancer C MV promoter
A G T C A T C G C T A T T A C C A T G G T G A T G C G G T T T T G G C A G T A C A T C A A T G G G C G T G G A T A G C G G T T T G A C T C A C G G G G A T T T C C A A G T
T C A G T A G C G A T A A T G G T A C C A C T A C G C C A A A A C C G T C A T G T A G T T A C C C G C A C C T A T C G C C A A A C T G A G T G C C C C T A A A G G T T C A
765
C MV promoter
C T C C A C C C C A T T G A C G T C A A T G G G A G T T T G T T T T G G C A C C A A A A T C A A C G G G A C T T T C C A A A A T G T C G T A A C A A C T C C G C C C C A T
G A G G T G G G G T A A C T G C A G T T A C C C T C A A A C A A A A C C G T G G T T T T A G T T G C C C T G A A A G G T T T T A C A G C A T T G T T G A G G C G G G G T A
850
C MV promoter
T G A C G C A A A T G G G C G G T A G G C G T G T A C G G T G G G A G G T C T A T A T A A G C A G A G C T C T C T G G C T A A C T A G A G A A C C C A C T G C T T A C T G
A C T G C G T T T A C C C G C C A T C C G C A C A T G C C A C C C T C C A G A T A T A T T C G T C T C G A G A G A C C G A T T G A T C T C T T G G G T G A C G A A T G A C
BamHIA f lI I HindII I A cc65I KpnINheI Bmt I
935
T7 promoter MC S
G C T T A T C G A A A T T A A T A C G A C T C A C T A T A G G G A G A C C C A A G C T G G C T A G C G T T T A A A C T T A A G C T T G G T A C C G A G C T C G G A T C C A
C G A A T A G C T T T A A T T A T G C T G A G T G A T A T C C C T C T G G G T T C G A C C G A T C G C A A A T T T G A A T T C G A A C C A T G G C T C G A G C C T A G G T
pcDNA3.1 Hygro(+).dna (Circular / 5597 bp)
Printed from SnapGene® Viewer: Tuesday, May 8, 2012 2:32:06 PM Page 3
BstXI Not I P aeR7IP spXIXhoI
TliI
XbaI
EcoO109IP spOMI
A paIEcoRVBstXI
1020
MC S
C T A G T C C A G T G T G G T G G A A T T C T G C A G A T A T C C A G C A C A G T G G C G G C C G C T C G A G T C T A G A G G G C C C G T T T A A A C C C G C T G A T C A
G A T C A G G T C A C A C C A C C T T A A G A C G T C T A T A G G T C G T G T C A C C G C C G G C G A G C T C A G A T C T C C C G G G C A A A T T T G G G C G A C T A G T
1105
bGH poly(A ) s ignal
G C C T C G A C T G T G C C T T C T A G T T G C C A G C C A T C T G T T G T T T G C C C C T C C C C C G T G C C T T C C T T G A C C C T G G A A G G T G C C A C T C C C A
C G G A G C T G A C A C G G A A G A T C A A C G G T C G G T A G A C A A C A A A C G G G G A G G G G G C A C G G A A G G A A C T G G G A C C T T C C A C G G T G A G G G T
1190
bGH poly(A ) s ignal
C T G T C C T T T C C T A A T A A A A T G A G G A A A T T G C A T C G C A T T G T C T G A G T A G G T G T C A T T C T A T T C T G G G G G G T G G G G T G G G G C A G G A
G A C A G G A A A G G A T T A T T T T A C T C C T T T A A C G T A G C G T A A C A G A C T C A T C C A C A G T A A G A T A A G A C C C C C C A C C C C A C C C C G T C C T
BbsI
1275
M A S E A E R T S
bGH poly(A ) s ignal
C A G C A A G G G G G A G G A T T G G G A A G A C A A T A G C A G G C A T G C T G G G G A T G C G G T G G G C T C T A T G G C T T C T G A G G C G G A A A G A A C C A G C
G T C G T T C C C C C T C C T A A C C C T T C T G T T A T C G T C C G T A C G A C C C C T A C G C C A C C C G A G A T A C C G A A G A C T C C G C C T T T C T T G G T C G
1360
W G S R G Y P H A P C S G A L S A A G V V V T R S V T A
f1 ori
T G G G G C T C T A G G G G G T A T C C C C A C G C G C C C T G T A G C G G C G C A T T A A G C G C G G C G G G T G T G G T G G T T A C G C G C A G C G T G A C C G C T A
A C C C C G A G A T C C C C C A T A G G G G T G C G C G G G A C A T C G C C G C G T A A T T C G C G C C G C C C A C A C C A C C A A T G C G C G T C G C A C T G G C G A T
1445
T L A S A L A P A P F A F F P S F L A T F A G F P R Q A L
f1 ori
C A C T T G C C A G C G C C C T A G C G C C C G C T C C T T T C G C T T T C T T C C C T T C C T T T C T C G C C A C G T T C G C C G G C T T T C C C C G T C A A G C T C T
G T G A A C G G T C G C G G G A T C G C G G G C G A G G A A A G C G A A A G A A G G G A A G G A A A G A G C G G T G C A A G C G G C C G A A A G G G G C A G T T C G A G A
1530
N R G L P L G F R F S A L R H L D P K K L D *
f1 ori
A A A T C G G G G G C T C C C T T T A G G G T T C C G A T T T A G T G C T T T A C G G C A C C T C G A C C C C A A A A A A C T T G A T T A G G G T G A T G G T T C A C G T
T T T A G C C C C C G A G G G A A A T C C C A A G G C T A A A T C A C G A A A T G C C G T G G A G C T G G G G T T T T T T G A A C T A A T C C C A C T A C C A A G T G C A
1615
f1 ori
A G T G G G C C A T C G C C C T G A T A G A C G G T T T T T C G C C C T T T G A C G T T G G A G T C C A C G T T C T T T A A T A G T G G A C T C T T G T T C C A A A C T G
T C A C C C G G T A G C G G G A C T A T C T G C C A A A A A G C G G G A A A C T G C A A C C T C A G G T G C A A G A A A T T A T C A C C T G A G A A C A A G G T T T G A C
1700
f1 ori
G A A C A A C A C T C A A C C C T A T C T C G G T C T A T T C T T T T G A T T T A T A A G G G A T T T T G C C G A T T T C G G C C T A T T G G T T A A A A A A T G A G C T
C T T G T T G T G A G T T G G G A T A G A G C C A G A T A A G A A A A C T A A A T A T T C C C T A A A A C G G C T A A A G C C G G A T A A C C A A T T T T T T A C T C G A
1785
f1 ori SV 40 promoter
G A T T T A A C A A A A A T T T A A C G C G A A T T A A T T C T G T G G A A T G T G T G T C A G T T A G G G T G T G G A A A G T C C C C A G G C T C C C C A G C A G G C A
C T A A A T T G T T T T T A A A T T G C G C T T A A T T A A G A C A C C T T A C A C A C A G T C A A T C C C A C A C C T T T C A G G G G T C C G A G G G G T C G T C C G T
pcDNA3.1 Hygro(+).dna (Circular / 5597 bp)
Printed from SnapGene® Viewer: Tuesday, May 8, 2012 2:32:06 PM Page 4
SexA I*
1870
SV 40 promoter
G A A G T A T G C A A A G C A T G C A T C T C A A T T A G T C A G C A A C C A G G T G T G G A A A G T C C C C A G G C T C C C C A G C A G G C A G A A G T A T G C A A A G
C T T C A T A C G T T T C G T A C G T A G A G T T A A T C A G T C G T T G G T C C A C A C C T T T C A G G G G T C C G A G G G G T C G T C C G T C T T C A T A C G T T T C
1955
SV 40 promoter
SV 40 ori
C A T G C A T C T C A A T T A G T C A G C A A C C A T A G T C C C G C C C C T A A C T C C G C C C A T C C C G C C C C T A A C T C C G C C C A G T T C C G C C C A T T C T
G T A C G T A G A G T T A A T C A G T C G T T G G T A T C A G G G C G G G G A T T G A G G C G G G T A G G G C G G G G A T T G A G G C G G G T C A A G G C G G G T A A G A
2040
SV 40 promoter
SV 40 ori
C C G C C C C A T G G C T G A C T A A T T T T T T T T A T T T A T G C A G A G G C C G A G G C C G C C T C T G C C T C T G A G C T A T T C C A G A A G T A G T G A G G A G
G G C G G G G T A C C G A C T G A T T A A A A A A A A T A A A T A C G T C T C C G G C T C C G G C G G A G A C G G A G A C T C G A T A A G G T C T T C A T C A C T C C T C
TspMIXmaI
SmaIBseRI StuI A vrI I
2125
M K K
* A K A F L E R S S T Y G N E S R I L V H H F L
1M K K
SV 40 promoter HygR
SV 40 ori
G C T T T T T T G G A G G C C T A G G C T T T T G C A A A A A G C T C C C G G G A G C T T G T A T A T C C A T T T T C G G A T C T G A T C A G C A C G T G A T G A A A A A
C G A A A A A A C C T C C G G A T C C G A A A A C G T T T T T C G A G G G C C C T C G A A C A T A T A G G T A A A A G C C T A G A C T A G T C G T G C A C T A C T T T T T
BsmBIP shA I
2210
P E L T A T S V E K F L I E K F D S V S D L M Q L S E G
R F E G R R R D L L K Q D F L E V A D G V Q H L E R L A
5 10 15 20 25 30P E L T A T S V E K F L I E K F D S V S D L M Q L S E G
HygR
G C C T G A A C T C A C C G C G A C G T C T G T C G A G A A G T T T C T G A T C G A A A A G T T C G A C A G C G T C T C C G A C C T G A T G C A G C T C T C G G A G G G C
C G G A C T T G A G T G G C G C T G C A G A C A G C T C T T C A A A G A C T A G C T T T T C A A G C T G T C G C A G A G G C T G G A C T A C G T C G A G A G C C T C C C G
2295
E E S R A F S F D V G G R G Y V L R V N S C A D G F Y K
F F R T S E A E I Y S P T S I D Q P Y I A A G I T E V F
35 40 45 50 55E E S R A F S F D V G G R G Y V L R V N S C A D G F Y K
HygR
G A A G A A T C T C G T G C T T T C A G C T T C G A T G T A G G A G G G C G T G G A T A T G T C C T G C G G G T A A A T A G C T G C G C C G A T G G T T T C T A C A A A G
C T T C T T A G A G C A C G A A A G T C G A A G C T A C A T C C T C C C G C A C C T A T A C A G G A C G C C C A T T T A T C G A C G C G G C T A C C A A A G A T G T T T C
2380
D R Y V Y R H F A S A A L P I P E V L D I G E F S E S L T
I T I N I P V K C R G R E R N R F H K V N P F E A L A Q G
60 65 70 75 80 85D R Y V Y R H F A S A A L P I P E V L D I G E F S E S L T
HygR
A T C G T T A T G T T T A T C G G C A C T T T G C A T C G G C C G C G C T C C C G A T T C C G G A A G T G C T T G A C A T T G G G G A A T T C A G C G A G A G C C T G A C
T A G C A A T A C A A A T A G C C G T G A A A C G T A G C C G G C G C G A G G G C T A A G G C C T T C A C G A A C T G T A A C C C C T T A A G T C G C T C T C G G A C T G
pcDNA3.1 Hygro(+).dna (Circular / 5597 bp)
Printed from SnapGene® Viewer: Tuesday, May 8, 2012 2:32:06 PM Page 5
BspMIBf uA I
2465
90 95 100 105 110 115Y C I S R R A Q G V T L Q D L P E T E L P A V L Q P V A
HygR
C T A T T G C A T C T C C C G C C G T G C A C A G G G T G T C A C G T T G C A A G A C C T G C C T G A A A C C G A A C T G C C C G C T G T T C T G C A G C C G G T C G C G
G A T A A C G T A G A G G G C G G C A C G T G T C C C A C A G T G C A A C G T T C T G G A C G G A C T T T G G C T T G A C G G G C G A C A A G A C G T C G G C C A G C G C
2550
2635
2720
2805
2890
G V D E E P P R P Q S T H L L L R A V E L P P M
pcDNA3.1 Hygro(+).dna (Circular / 5597 bp)
Printed from SnapGene® Viewer: Tuesday, May 8, 2012 2:32:06 PM Page 6
SacII
2975
S P R L R A Y M L R I G L D Q L Y Q S L V D G N F D D A
260 265 270 275 280 285S P R L R A Y M L R I G L D Q L Y Q S L V D G N F D D A
HygR
A T C G C C G C G G C T C C G G G C G T A T A T G C T C C G C A T T G G T C T T G A C C A A C T C T A T C A G A G C T T G G T T G A C G G C A A T T T C G A T G A T G C A
T A G C G G C G C C G A G G C C C G C A T A T A C G A G G C G T A A C C A G A A C T G G T T G A G A T A G T C T C G A A C C A A C T G C C G T T A A A G C T A C T A C G T
3060
A W A Q G R C D A I V R S G A G T V G R T Q I A R R S A
290 295 300 305 310A W A Q G R C D A I V R S G A G T V G R T Q I A R R S A
HygR
G C T T G G G C G C A G G G T C G A T G C G A C G C A A T C G T C C G A T C C G G A G C C G G G A C T G T C G G G C G T A C A C A A A T C G C C C G C A G A A G C G C G G
C G A A C C C G C G T C C C A G C T A C G C T G C G T T A G C A G G C T A G G C C T C G G C C C T G A C A G C C C G C A T G T G T T T A G C G G G C G T C T T C G C G C C
3145
A V W T D G C V E V L A D S G N R R P S T R P R A K E *
315 320 325 330 335 340A V W T D G C V E V L A D S G N R R P S T R P R A K E *
HygR
C C G T C T G G A C C G A T G G C T G T G T A G A A G T A C T C G C C G A T A G T G G A A A C C G A C G C C C C A G C A C T C G T C C G A G G G C A A A G G A A T A G C A
G G C A G A C C T G G C T A C C G A C A C A T C T T C A T G A G C G G C T A T C A C C T T T G G C T G C G G G G T C G T G A G C A G G C T C C C G T T T C C T T A T C G T
P f oI
3230C G T G C T A C G A G A T T T C G A T T C C A C C G C C G C C T T C T A T G A A A G G T T G G G C T T C G G A A T C G T T T T C C G G G A C G C C G G C T G G A T G A T C
G C A C G A T G C T C T A A A G C T A A G G T G G C G G C G G A A G A T A C T T T C C A A C C C G A A G C C T T A G C A A A A G G C C C T G C G G C C G A C C T A C T A G
3315
SV 40 poly(A ) s ignal
C T C C A G C G C G G G G A T C T C A T G C T G G A G T T C T T C G C C C A C C C C A A C T T G T T T A T T G C A G C T T A T A A T G G T T A C A A A T A A A G C A A T A
G A G G T C G C G C C C C T A G A G T A C G A C C T C A A G A A G C G G G T G G G G T T G A A C A A A T A A C G T C G A A T A T T A C C A A T G T T T A T T T C G T T A T
BsmI
3400
SV 40 poly(A ) s ignal
G C A T C A C A A A T T T C A C A A A T A A A G C A T T T T T T T C A C T G C A T T C T A G T T G T G G T T T G T C C A A A C T C A T C A A T G T A T C T T A T C A T G T
C G T A G T G T T T A A A G T G T T T A T T T C G T A A A A A A A G T G A C G T A A G A T C A A C A C C A A A C A G G T T T G A G T A G T T A C A T A G A A T A G T A C A
BstZ17I
3485
M13 rev
C T G T A T A C C G T C G A C C T C T A G C T A G A G C T T G G C G T A A T C A T G G T C A T A G C T G T T T C C T G T G T G A A A T T G T T A T C C G C T C A C A A T T
G A C A T A T G G C A G C T G G A G A T C G A T C T C G A A C C G C A T T A G T A C C A G T A T C G A C A A A G G A C A C A C T T T A A C A A T A G G C G A G T G T T A A
3570C C A C A C A A C A T A C G A G C C G G A A G C A T A A A G T G T A A A G C C T G G G G T G C C T A A T G A G T G A G C T A A C T C A C A T T A A T T G C G T T G C G C T
G G T G T G T T G T A T G C T C G G C C T T C G T A T T T C A C A T T T C G G A C C C C A C G G A T T A C T C A C T C G A T T G A G T G T A A T T A A C G C A A C G C G A
3655C A C T G C C C G C T T T C C A G T C G G G A A A C C T G T C G T G C C A G C T G C A T T A A T G A A T C G G C C A A C G C G C G G G G A G A G G C G G T T T G C G T A T
G T G A C G G G C G A A A G G T C A G C C C T T T G G A C A G C A C G G T C G A C G T A A T T A C T T A G C C G G T T G C G C G C C C C T C T C C G C C A A A C G C A T A
BspQISapI
3740T G G G C G C T C T T C C G C T T C C T C G C T C A C T G A C T C G C T G C G C T C G G T C G T T C G G C T G C G G C G A G C G G T A T C A G C T C A C T C A A A G G C G
A C C C G C G A G A A G G C G A A G G A G C G A G T G A C T G A G C G A C G C G A G C C A G C A A G C C G A C G C C G C T C G C C A T A G T C G A G T G A G T T T C C G C
pcDNA3.1 Hygro(+).dna (Circular / 5597 bp)
Printed from SnapGene® Viewer: Tuesday, May 8, 2012 2:32:06 PM Page 7
P ciI
3825G T A A T A C G G T T A T C C A C A G A A T C A G G G G A T A A C G C A G G A A A G A A C A T G T G A G C A A A A G G C C A G C A A A A G G C C A G G A A C C G T A A A A
C A T T A T G C C A A T A G G T G T C T T A G T C C C C T A T T G C G T C C T T T C T T G T A C A C T C G T T T T C C G G T C G T T T T C C G G T C C T T G G C A T T T T
3910
ori
A G G C C G C G T T G C T G G C G T T T T T C C A T A G G C T C C G C C C C C C T G A C G A G C A T C A C A A A A A T C G A C G C T C A A G T C A G A G G T G G C G A A A
T C C G G C G C A A C G A C C G C A A A A A G G T A T C C G A G G C G G G G G G A C T G C T C G T A G T G T T T T T A G C T G C G A G T T C A G T C T C C A C C G C T T T
3995
ori
C C C G A C A G G A C T A T A A A G A T A C C A G G C G T T T C C C C C T G G A A G C T C C C T C G T G C G C T C T C C T G T T C C G A C C C T G C C G C T T A C C G G A
G G G C T G T C C T G A T A T T T C T A T G G T C C G C A A A G G G G G A C C T T C G A G G G A G C A C G C G A G A G G A C A A G G C T G G G A C G G C G A A T G G C C T
4080
ori
T A C C T G T C C G C C T T T C T C C C T T C G G G A A G C G T G G C G C T T T C T C A T A G C T C A C G C T G T A G G T A T C T C A G T T C G G T G T A G G T C G T T C
A T G G A C A G G C G G A A A G A G G G A A G C C C T T C G C A C C G C G A A A G A G T A T C G A G T G C G A C A T C C A T A G A G T C A A G C C A C A T C C A G C A A G
4165
ori
G C T C C A A G C T G G G C T G T G T G C A C G A A C C C C C C G T T C A G C C C G A C C G C T G C G C C T T A T C C G G T A A C T A T C G T C T T G A G T C C A A C C C
C G A G G T T C G A C C C G A C A C A C G T G C T T G G G G G G C A A G T C G G G C T G G C G A C G C G G A A T A G G C C A T T G A T A G C A G A A C T C A G G T T G G G
4250
ori
G G T A A G A C A C G A C T T A T C G C C A C T G G C A G C A G C C A C T G G T A A C A G G A T T A G C A G A G C G A G G T A T G T A G G C G G T G C T A C A G A G T T C
C C A T T C T G T G C T G A A T A G C G G T G A C C G T C G T C G G T G A C C A T T G T C C T A A T C G T C T C G C T C C A T A C A T C C G C C A C G A T G T C T C A A G
4335
ori
T T G A A G T G G T G G C C T A A C T A C G G C T A C A C T A G A A G A A C A G T A T T T G G T A T C T G C G C T C T G C T G A A G C C A G T T A C C T T C G G A A A A A
A A C T T C A C C A C C G G A T T G A T G C C G A T G T G A T C T T C T T G T C A T A A A C C A T A G A C G C G A G A C G A C T T C G G T C A A T G G A A G C C T T T T T
4420
ori
G A G T T G G T A G C T C T T G A T C C G G C A A A C A A A C C A C C G C T G G T A G C G G T T T T T T T G T T T G C A A G C A G C A G A T T A C G C G C A G A A A A A A
C T C A A C C A T C G A G A A C T A G G C C G T T T G T T T G G T G G C G A C C A T C G C C A A A A A A A C A A A C G T T C G T C G T C T A A T G C G C G T C T T T T T T
4505
ori
A G G A T C T C A A G A A G A T C C T T T G A T C T T T T C T A C G G G G T C T G A C G C T C A G T G G A A C G A A A A C T C A C G T T A A G G G A T T T T G G T C A T G
T C C T A G A G T T C T T C T A G G A A A C T A G A A A A G A T G C C C C A G A C T G C G A G T C A C C T T G C T T T T G A G T G C A A T T C C C T A A A A C C A G T A C
4590A G A T T A T C A A A A A G G A T C T T C A C C T A G A T C C T T T T A A A T T A A A A A T G A A G T T T T A A A T C A A T C T A A A G T A T A T A T G A G T A A A C T T
T C T A A T A G T T T T T C C T A G A A G T G G A T C T A G G A A A A T T T A A T T T T T A C T T C A A A A T T T A G T T A G A T T T C A T A T A T A C T C A T T T G A A
A hdI
4675
* W H K I L S A G I E A I Q R N R E D M T A Q S G
285 280 275 270 265* W H K I L S A G I E A I Q R N R E D M T A Q S G
A mpR
G G T C T G A C A G T T A C C A A T G C T T A A T C A G T G A G G C A C C T A T C T C A G C G A T C T G T C T A T T T C G T T C A T C C A T A G T T G C C T G A C T C C C
C C A G A C T G T C A A T G G T T A C G A A T T A G T C A C T C C G T G G A T A G A G T C G C T A G A C A G A T A A A G C A A G T A G G T A T C A A C G G A C T G A G G G
pcDNA3.1 Hygro(+).dna (Circular / 5597 bp)
Printed from SnapGene® Viewer: Tuesday, May 8, 2012 2:32:06 PM Page 8
4760
M I P R D P R S P A
T T Y I V V I R S P K G D P G L A A I I G R S G R E G A
260 255 250 245 240 235T T Y I V V I R S P K G D P G L A A I I G R S G R E G A
A mpR
C G T C G T G T A G A T A A C T A C G A T A C G G G A G G G C T T A C C A T C T G G C C C C A G T G C T G C A A T G A T A C C G C G A G A C C C A C G C T C A C C G G C T
G C A G C A C A T C T A T T G A T G C T A T G C C C T C C C G A A T G G T A G A C C G G G G T C A C G A C G T T A C T A T G G C G C T C T G G G T G C G A G T G G C C G A
4845
P D L S A I N Q P A G R A E R R S G P A T L S A S I Q S
G S K D A I F W G A P L A S R L L P G A V K D A E M W D I
230 225 220 215 210G S K D A I F W G A P L A S R L L P G A V K D A E M W D I
A mpR
C C A G A T T T A T C A G C A A T A A A C C A G C C A G C C G G A A G G G C C G A G C G C A G A A G T G G T C C T G C A A C T T T A T C C G C C T C C A T C C A G T C T A
G G T C T A A A T A G T C G T T A T T T G G T C G G T C G G C C T T C C C G G C T C G C G T C T T C A C C A G G A C G T T G A A A T A G G C G G A G G T A G G T C A G A T
FspI
4930
I N C C R E A R V S S S P V N S L R N V V A I A T G I V V
L Q Q R S A L T L L E G T L L K R L T T A M A V P M T T
205 200 195 190 185 180L Q Q R S A L T L L E G T L L K R L T T A M A V P M T T
A mpR
T T A A T T G T T G C C G G G A A G C T A G A G T A A G T A G T T C G C C A G T T A A T A G T T T G C G C A A C G T T G T T G C C A T T G C T A C A G G C A T C G T G G T
A A T T A A C A A C G G C C C T T C G A T C T C A T T C A T C A A G C G G T C A A T T A T C A A A C G C G T T G C A A C A A C G G T A A C G A T G T C C G T A G C A C C A
5015
S R S S F G M A S F S S G S Q R S R R V T *D R E D N P I A E N L E P E W R D L R T V H D G M N H L
175 170 165 160 155 150D R E D N P I A E N L E P E W R D L R T V H D G M N H L
A mpR
G T C A C G C T C G T C G T T T G G T A T G G C T T C A T T C A G C T C C G G T T C C C A A C G A T C A A G G C G A G T T A C A T G A T C C C C C A T G T T G T G C A A A
C A G T G C G A G C A G C A A A C C A T A C C G A A G T A A G T C G A G G C C A A G G G T T G C T A G T T C C G C T C A A T G T A C T A G G G G G T A C A A C A C G T T T
5100
F A T L E K P G G I T T L L L N A A T N D S M T I A A S C
145 140 135 130 125F A T L E K P G G I T T L L L N A A T N D S M T I A A S C
A mpR
A A A G C G G T T A G C T C C T T C G G T C C T C C G A T C G T T G T C A G A A G T A A G T T G G C C G C A G T G T T A T C A C T C A T G G T T A T G G C A G C A C T G C
T T T C G C C A A T C G A G G A A G C C A G G A G G C T A G C A A C A G T C T T C A T T C A A C C G G C G T C A C A A T A G T G A G T A C C A A T A C C G T C G T G A C G
5185
L E R V T M G D T L H K E T V P S Y E V L D N Q S Y H I
120 115 110 105 100 95L E R V T M G D T L H K E T V P S Y E V L D N Q S Y H I
A mpR
A T A A T T C T C T T A C T G T C A T G C C A T C C G T A A G A T G C T T T T C T G T G A C T G G T G A G T A C T C A A C C A A G T C A T T C T G A G A A T A G T G T A T
T A T T A A G A G A A T G A C A G T A C G G T A G G C A T T C T A C G A A A A G A C A C T G A C C A C T C A T G A G T T G G T T C A G T A A G A C T C T T A T C A C A T A
pcDNA3.1 Hygro(+).dna (Circular / 5597 bp)
Printed from SnapGene® Viewer: Tuesday, May 8, 2012 2:32:06 PM Page 9
5270
R R G L Q E Q G A D I R S L V A G C L L V K F T S M M P
90 85 80 75 70 65R R G L Q E Q G A D I R S L V A G C L L V K F T S M M P
A mpR
G C G G C G A C C G A G T T G C T C T T G C C C G G C G T C A A T A C G G G A T A A T A C C G C G C C A C A T A G C A G A A C T T T A A A A G T G C T C A T C A T T G G A
C G C C G C T G G C T C A A C G A G A A C G G G C C G C A G T T A T G C C C T A T T A T G G C G C G G T G T A T C G T C T T G A A A T T T T C A C G A G T A G T A A C C T
5355
F R E E P R F S E L I K G S N L D L E I Y G V R A G L Q D
60 55 50 45 40F R E E P R F S E L I K G S N L D L E I Y G V R A G L Q D
A mpR
A A A C G T T C T T C G G G G C G A A A A C T C T C A A G G A T C T T A C C G C T G T T G A G A T C C A G T T C G A T G T A A C C C A C T C G T G C A C C C A A C T G A T
T T T G C A A G A A G C C C C G C T T T T G A G A G T T C C T A G A A T G G C G A C A A C T C T A G G T C A A G C T A C A T T G G G T G A G C A C G T G G G T T G A C T A
5440
E A D K V K V L T E P H A F V P L C F A A F F P I L A V
35 30 25E A D K V K V L T E P H
20 15 10
s ignal sequenceA F V P L C F A A F F P I L A V
A mpR
C T T C A G C A T C T T T T A C T T T C A C C A G C G T T T C T G G G T G A G C A A A A A C A G G A A G G C A A A A T G C C G C A A A A A A G G G A A T A A G G G C G A C
G A A G T C G T A G A A A A T G A A A G T G G T C G C A A A G A C C C A C T C G T T T T T G T C C T T C C G T T T T A C G G C G T T T T T T C C C T T A T T C C C G C T G
SspI
5525
R F H Q I S M
5 1
s ignal sequenceR F H Q I S M
A mpRA mpR promoter
A C G G A A A T G T T G A A T A C T C A T A C T C T T C C T T T T T C A A T A T T A T T G A A G C A T T T A T C A G G G T T A T T G T C T C A T G A G C G G A T A C A T A
T G C C T T T A C A A C T T A T G A G T A T G A G A A G G A A A A A G T T A T A A T A A C T T C G T A A A T A G T C C C A A T A A C A G A G T A C T C G C C T A T G T A T
SgrDI
3 ʹ
5 ʹ5597
A mpR promoter
T T T G A A T G T A T T T A G A A A A A T A A A C A A A T A G G G G T T C C G C G C A C A T T T C C C C G A A A A G T G C C A C C T G A C G T C
A A A C T T A C A T A A A T C T T T T T A T T T G T T T A T C C C C A A G G C G C G T G T A A A G G G G C T T T T C A C G G T G G A C T G C A G
pcDNA3.1 Hygro(+).dna (Circular / 5597 bp)
Enzymes SitesA cc65I 1 917A f lI I 1 908A hdI 1 4674A paI 1 1001A siSI 1 2482A vrI I 1 2054BamHI 1 929BbsI 1 1217Bf uA I 1 2431BglI I 1 12Bmt I 1 899Bpu10I 1 180BseRI 1 2050BsmI 1 3353BsmBI 1 2186BspMI 1 2431BspQI 1 3668BstXI 2 948 974BstZ17I 1 3405EcoO109I 1 997EcoRV 1 964FspI 1 4896HindII I 1 911KpnI 1 921Mf eI 1 161MluI 1 228NheI 1 895Not I 1 979NruI 1 208PaeR7I 1 985P ciI 1 3784P f oI 1 3208P shA I 1 2145P spOMI 1 997P spXI 1 985RsrI I 1 2526SacII 1 2898SapI 1 3668SexA I 1* 1821*SgrDI 1 5595SmaI 1 2077SnaBI 1 590SspI 1 5478StuI 1 2053TliI 1 985TspMI 1 2075XbaI 1 991XhoI 1 985XmaI 1 2075
Printed from SnapGene® Viewer: Tuesday, May 8, 2012 2:32:08 PM Page 10
pcDNA3.1 Hygro(+).dna (Circular / 5597 bp)
Printed from SnapGene® Viewer: Tuesday, May 8, 2012 2:32:08 PM Page 11
Feature Location Size (bp) Type
✓ CMV enhancer 235 .. 614 380 enhancer
/note = human cytomegalovirus immediate early enhancer
✓ CMV promoter 615 .. 818 204 promoter
/note = human cytomegalovirus (C MV ) immediate early promoter
✓ T7 promoter 863 .. 881 19 promoter
/note = promoter for bac teriophage T7 RNA polymerase
✓ MCS 895 .. 1002 108 misc_feature
/note = multiple c loning s ite
✓ bGH poly(A ) signal 1028 .. 1252 225 polyA _s ignal
/note = bovine growth hormone polyadenylation s ignal
✓ f 1 ori 1298 .. 1726 429 rep_origin
/direc tion = RIGHT
/note = f1 bac teriophage origin of replication; arrow indicates direc tion of (+) s trand synthes is
✓ SV40 promoter 1740 .. 2069 330 promoter
/note = SV 40 enhancer and early promoter
✓ SV40 ori 1920 .. 2055 136 rep_origin
/note = SV 40 origin of replication
✓ HygR 2118 .. 3143 1026 C DS
/gene = aph(4)-Ia
/produc t = aminoglycos ide phosphotrans ferase from E. coli
/note = confers res is tance to hygromyc in
/trans lation = MKKPELTA TSV EKFLIEKFDSV SDLMQ LSEGEESRA FSFDV GGRGYV LRV NSC A DGFYKDRYV YRHFA SA A LP IPEV LDIGEFSESLTYC ISRRA Q GV TLQ DLPETELPA V LQ PV A EA MDA IA A A DLSQ TSGFGPFGPQ GIGQ YTTWRDFIC A IA DPHV YHWQ TV MDDTV SASV A Q A LDELMLWA EDC PEV RHLV HA DFGSNNV LTDNGRITA V IDWSEA MFGDSQ YEV A NIFFWRPWLA C MEQ Q TRYFERRHPELA GSPRLRA YMLRIGLDQ LYQ SLV DGNFDDA A WA Q GRC DA IV RSGA GTV GRTQ IA RRSA A V WTDGC V EV LA DSGNRRPSTRPRA KE*341 amino ac ids = 38 .0 kDa
✓ SV40 poly(A ) signal 3273 .. 3394 122 polyA _s ignal
/note = SV 40 polyadenylation s ignal
✓ M13 rev 3443 .. 3459 17 primer_bind
/note = common sequenc ing primer, one of multiple s imilar variants
✓ ori 3845 .. 4430 586 rep_origin
/direc tion = LEFT
/note = high-copy-number C olE1/pMB1/pBR322/pUC origin of replication
✓ A mpR 4601 .. 5461 861 C DS
2 segments
/gene = bla
/produc t = β-lac tamase
/note = confers res is tance to ampic illin, carbenic illin, and related antibiotic s
/trans lation = MSIQ HFRV A LIP FFA A FC LPV FA ,HPETLV KV KDA EDQ LGA RV GYIELDLNSGKILESFRPEERFPMMSTFKV LLC GA V LSRIDA GQ EQ LGRRIHYSQ NDLV EYSPV TEKHLTDGMTV RELC SA A ITMSDNTA A NLLLTT IGGPKELTA FLHNMGDHV TRLDRWEPELNEA IPNDERDTTMPV A MA TTLRKLLTGELLTLA SRQ Q LIDWMEA DKV A GPLLRSA LPA GWFIA DKSGA GERGSRGIIA A LGPDGKPSRIV V IYTTGSQ A TMDERNRQ IA EIGA SLIKHW*286 amino ac ids = 31 .6 kDa
pcDNA3.1 Hygro(+).dna (Circular / 5597 bp)
Printed from SnapGene® Viewer: Tuesday, May 8, 2012 2:32:08 PM Page 12
Feature Location Size (bp) Type
✓ A mpR promoter 5462 .. 5566 105 promoter
/gene = bla
pcDNA3.1 Hygro(+).dna (Circular / 5597 bp)
Descript ion: Mammalian express ion vec tor with the C MV promoter. The MC S is in the forward (+) orientation.Created: Tuesday, Jul 31 , 2012
Last Modif ied: Saturday, Jan 5 , 2013A ccession Number:
Code Number:Sequence A uthor: Invitrogen (Life Technologies)
DNA Type: Synthetic DNALaboratory Host Organism: Mammalian C ells
Bact erial Transf ormat ion St rain: Unspec ifiedDam+ Dcm+ EcoKI+
Comment s: The c loned gene must inc lude a Kozak sequence and s tart codon.
Ref erences:
Printed from SnapGene® Viewer: Tuesday, May 8, 2012 2:32:08 PM Page 13