pcdna™3.1/hygro(+) - tsv-co.com plasmid/pcdna3.1... · pcdna3.1 hygro(+).dna (circular / 5597 bp)...

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