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

13
Unique Cutters Bold Sequence: 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) PciI (3784) BspQI - SapI (3668) Bst Z17I (3405) BsmI (3353) PfoI (3208) SacII (2898) BglII (12) Mf eI (161) Bpu10I (180) NruI (208) MluI (228) SnaBI (590) NheI (895) BmtI (899) AflII (908) HindIII (911) A cc65I (917) KpnI (921) BamHI (929) BstXI (948) EcoRV (964) BstXI (974) NotI (979) PaeR7I - PspXI - TliI - XhoI (985) XbaI (991) EcoO109I - PspOMI (997) A paI (1001) BbsI (1217) SexAI* (1821) BseRI (2050) St uI (2053) AvrII (2054) TspMI - XmaI (2075) SmaI (2077) P shA I (2145) BsmBI (2186) BfuAI - BspMI (2431) A siSI (2482) RsrII (2526) pcDNA™3.1/ Hygro(+) 5597 bp

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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