pEASYBlunt clonning vector 载体说明书WORD版.docx
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pEASYBluntclonningvector载体说明书WORD版
pEASY-Blunt
Completesequence
AGCGCCCAATACGCAAACCGCCTCTCCCCGCGCGTTGGCCGATTCATTAATGCAGCTGGCACGACAGGTTTCCCGACTGGAAAGCGGGCAGTGAGCGCAACGCAATTAATGTGAGTTAGCTCACTCATTAGGCACCCCAGGCTTTACACTTTATGCTTCCGGCTCGTATGTTGTGTGGAATTGTGAGCGGATAACAATTTCACACAGGAAACAGCTATGACCATGATTACGCCAAGCTTGGTACCGAGCTCGGATCCACTAGTAACGGCCGCCAGTGTGCTGGAATTGCCCTTAAGGGCAATTCTGCAGATATCCATCACACTGGCGGCCGCTCGAGCATGCATCTAGAGGGCCCAATTCGCCCTATAGTGAGTCGTATTACAATTCACTGGCCGTCGTTTTACAACGTCGTGACTGGGAAAACCCTGGCGTTACCCAACTTAATCGCCTTGCAGCACATCCCCCTTTCGCCAGCTGGCGTAATAGCGAAGAGGCCCGCACCGATCGCCCTTCCCAACAGTTGCGCAGCCTGAATGGCGAATGGACGCGCCCTGTAGCGGCGCATTAAGCGCGGCGGGTGTGGTGGTTACGCGCAGCGTGACCGCTACACTTGCCAGCGCCCTAGCGCCCGCTCCTTTCGCTTTCTTCCCTTCCTTTCTCGCCACGTTCGCCGGCTTTCCCCGTCAAGCTCTAAATCGGGGGCTCCCTTTAGGGTTCCGATTTAGTGCTTTACGGCACCTCGACCCCAAAAAACTTGATTAGGGTGATGGTTCACGTAGTGGGCCATCGCCCTGATAGACGGTTTTTCGCCCTTTGACGTTGGAGTCCACGTTCTTTAATAGTGGACTCTTGTTCCAAACTGGAACAACACTCAACCCTATCTCGGTCTATTCTTTTGATTTATAAGGGATTTTGCCGATTTCGGCCTATTGGTTAAAAAATGAGCTGATTTAACAAAAATTTAACGCGAATTTTAACAAAATTCAGGGCGCAAGGGCTGCTAAAGGAAGCGGAACACGTAGAAAGCCAGTCCGCAGAAACGGTGCTGACCCCGGATGAATGTCAGCTACTGGGCTATCTGGACAAGGGAAAACGCAAGCGCAAAGAGAAAGCAGGTAGCTTGCAGTGGGCTTACATGGCGATAGCTAGACTGGGCGGTTTTATGGACAGCAAGCGAACCGGAATTGCCAGCTGGGGCGCCCTCTGGTAAGGTTGGGAAGCCCTGCAAAGTAAACTGGATGGCTTTCTTGCCGCCAAGGATCTGATGGCGCAGGGGATCAAGATCTGATCAAGAGACAGGATGAGGATCGTTTCGCATGATTGAACAAGATGGATTGCACGCAGGTTCTCCGGCCGCTTGGGTGGAGAGGCTATTCGGCTATGACTGGGCACAACAGACAATCGGCTGCTCTGATGCCGCCGTGTTCCGGCTGTCAGCGCAGGGGCGCCCGGTTCTTTTTGTCAAGACCGACCTGTCCGGTGCCCTGAATGAACTGCAGGACGAGGCAGCGCGGCTATCGTGGCTGGCCACGACGGGCGTTCCTTGCGCAGCTGTGCTCGACGTTGTCACTGAAGCGGGAAGGGACTGGCTGCTATTGGGCGAAGTGCCGGGGCAGGATCTCCTGTCATCCCACCTTGCTCCTGCCGAGAAAGTATCCATCATGGCTGATGCAATGCGGCGGCTGCATACGCTTGATCCGGCTACCTGCCCATTCGACCACCAAGCGAAACATCGCATCGAGCGAGCACGTACTCGGATGGAAGCCGGTCTTGTCGATCAGGATGATCTGGACGAAGAGCATCAGGGGCTCGCGCCAGCCGAACTGTTCGCCAGGCTCAAGGCGCGCATGCCCGACGGCGAGGATCTCGTCGTGACCCATGGCGATGCCTGCTTGCCGAATATCATGGTGGAAAATGGCCGCTTTTCTGGATTCATCGACTGTGGCCGGCTGGGTGTGGCGGACCGCTATCAGGACATAGCGTTGGCTACCCGTGATATTGCTGAAGAGCTTGGCGGCGAATGGGCTGACCGCTTCCTCGTGCTTTACGGTATCGCCGCTCCCGATTCGCAGCGCATCGCCTTCTATCGCCTTCTTGACGAGTTCTTCTGAATTGAAAAAGGAAGAGTATGAGTATTCAACATTTCCGTGTCGCCCTTATTCCCTTTTTTGCGGCATTTTGCCTTCCTGTTTTTGCTCACCCAGAAACGCTGGTGAAAGTAAAAGATGCTGAAGATCAGTTGGGTGCACGAGTGGGTTACATCGAACTGGATCTCAACAGCGGTAAGATCCTTGAGAGTTTTCGCCCCGAAGAACGTTTTCCAATGATGAGCACTTTTAAAGTTCTGCTATGTGGCGCGGTATTATCCCGTATTGACGCCGGGCAAGAGCAACTCGGTCGCCGCATACACTATTCTCAGAATGACTTGGTTGAGTACTCACCAGTCACAGAAAAGCATCTTACGGATGGCATGACAGTAAGAGAATTATGCAGTGCTGCCATAACCATGAGTGATAACACTGCGGCCAACTTACTTCTGACAACGATCGGAGGACCGAAGGAGCTAACCGCTTTTTTGCACAACATGGGGGATCATGTAACTCGCCTTGATCGTTGGGAACCGGAGCTGAATGAAGCCATACCAAACGACGAGCGTGACACCACGATGCCTGTAGCAATGGCAACAACGTTGCGCAAACTATTAACTGGCGAACTACTTACTCTAGCTTCCCGGCAACAATTAATAGACTGGATGGAGGCGGATAAAGTTGCAGGACCACTTCTGCGCTCGGCCCTTCCGGCTGGCTGGTTTATTGCTGATAAATCTGGAGCCGGTGAGCGTGGGTCTCGCGGTATCATTGCAGCACTGGGGCCAGATGGTAAGCCCTCCCGTATCGTAGTTATCTACACGACGGGGAGTCAGGCAACTATGGATGAACGAAATAGACAGATCGCTGAGATAGGTGCCTCACTGATTAAGCATTGGTAACTGTCAGACCAAGTTTACTCATATATACTTTAGATTGATTTAAAACTTCATTTTTAATTTAAAAGGATCTAGGTGAAGATCCTTTTTGATAATCTCATGACCAAAATCCCTTAACGTGAGTTTTCGTTCCACTGAGCGTCAGACCCCGTAGAAAAGATCAAAGGATCTTCTTGAGATCCTTTTTTTCTGCGCGTAATCTGCTGCTTGCAAACAAAAAAACCACCGCTACCAGCGGTGGTTTGTTTGCCGGATCAAGAGCTACCAACTCTTTTTCCGAAGGTAACTGGCTTCAGCAGAGCGCAGATACCAAATACTGTTCTTCTAGTGTAGCCGTAGTTAGGCCACCACTTCAAGAACTCTGTAGCACCGCCTACATACCTCGCTCTGCTAATCCTGTTACCAGTGGCTGCTGCCAGTGGCGATAAGTCGTGTCTTACCGGGTTGGACTCAAGACGATAGTTACCGGATAAGGCGCAGCGGTCGGGCTGAACGGGGGGTTCGTGCACACAGCCCAGCTTGGAGCGAACGACCTACACCGAACTGAGATACCTACAGCGTGAGCTATGAGAAAGCGCCACGCTTCCCGAAGGGAGAAAGGCGGACAGGTATCCGGTAAGCGGCAGGGTCGGAACAGGAGAGCGCACGAGGGAGCTTCCAGGGGGAAACGCCTGGTATCTTTATAGTCCTGTCGGGTTTCGCCACCTCTGACTTGAGCGTCGATTTTTGTGATGCTCGTCAGGGGGGCGGAGCCTATGGAAAAACGCCAGCAACGCGGCCTTTTTACGGTTCCTGGCCTTTTGCTGGCCTTTTGCTCACATGTTCTTTCCTGCGTTATCCCCTGATTCTGTGGATAACCGTATTACCGCCTTTGAGTGAGCTGATACCGCTCGCCGCAGCCGAACGACCGAGCGCAGCGAGTCAGTGAGCGAGGAAGCGGAAG
LacZαfragment:
bases1-548
M13reverseprimingsite:
bases205-221
Multiplecloningsite:
bases234-358
T7promoter/primingsite:
bases364-381
M13Forward(-20)primingsite:
bases389-404
f1origin:
bases546-983
KanamycinresistanceORF:
bases1317-2111
AmpicillinresistanceORF:
bases2129-2989
pUCorigin:
bases3134-3807
pEASY-Blunt
RestrictionMap
Enzyme
#ofcuts
positions
Acc65I
(1)
ggtacc
246
AceIII(3)
cagctc
949,2630,3870
AciI(47)
ccgc
88,191,329,561,575,1014,1159,1515,1579,1680
AclI
(2)
aacgtt
2319,2692
AcuI(4)
ctgaag
3283,1576,2008,2235
AflII
(1)
cttaag
297
AflIII
(2)
acrygt
1021,3810
AhdI
(1)
gacnnnnngtc
2922
AloI
(1)
gaacnnnnnntcc
835
AluI(22)
agct
58,122,217,239,251,477,691,948,1069,1122
AlwI(16)
ggatc
258,1700,2292,3072,3169,257,1263,1280,1310,1621
AlwNI
(2)
cagnnnctg
1073,3399
ApaBI
(1)
gcannnnntgc
1002
ApaI
(1)
gggccc
356
ApaLI
(2)
gtgcac
2250,3496
ApoI(3)
raatty
964,975,986
AseI(3)
attaat
52,111,2746
AvaI
(1)
cycgrg
338
AvaII(3)
ggwcc
1966,2557,2779
BamHI
(1)
ggatcc
258
BanI(7)
ggyrcc
137,246,740,1202,1450,1485,2969
BanII(4)
grgcyc
252,356,706,1812
BbeI
(2)
ggcgcc
1202,1450
BbvI(18)
gcagc
1002,1410,1594,1687,2500,3194,3400,57,457,530
BccI(11)
ccatc
771,1243,1269,1334,1762,2470,2757,2881,319,789
BceAI(6)
acggc
397,1424,3324,270,737,1860
BcefI(6)
acggc
397,1424,3324,270,737,1860
BcgI
(1)
cgannnnnntgc
2401
BciVI
(2)
gtatcc
1659,3601
BclI
(1)
tgatca
1292
BfaI(7)
ctag
263,349,626,1150,2727,3062,3315
BfrBI
(1)
atgcat
345
BglI
(2)
gccnnnnnggc
539,2803
BglII
(1)
agatct
1287
Bme1580I(5)
gkgcmc
356,1393,1486,2250,3496
BmrI(5)
actggg
420,1075,1156,1390,2872
BpmI
(1)
ctggag
2832
BpuEI(5)
cttgag
1842,3442,2297,3165,3704
BsaAI(3)
yacgtr
779,1022,1753
BsaBI(3)
gatnnnnatc
319,1286,1310
BsaHI(3)
grcgyc
1202,1450,2381
BsaI
(1)
ggtctc
2850
BsaJI(7)
ccnngg
142,430,1056,1260,1614,1883,3650
BsaWI(5)
wccggw
1184,1482,2626,3457,3604
BsaXI
(2)
acnnnnnctcc
833,28
BscAI(11)
gcatc
1418,1673,1889,2230,2670,3722,346,1740,1804,2079
BseMII(4)
ctcag
2960,3126,3535,2420
BseYI(3)
cccagc
1197,1955,3506
BsiEI(8)
cgrycg
272,332,508,1357,2401,2550,3473,3897
BsiHKAI(6)
gwgcwc
252,1560,1750,2250,2335,3496
BslI(9)
ccnnnnnnngg
39,561,887,1431,1975,3331,3610,3776,3794
BsmAI
(2)
gtctc
1298,2850
BsmFI
(1)
gggac
1587
Bsp1286I(11)
gdgchc
252,356,706,1393,1486,1560,1750,1812,2250,2335
Bsp24I
(1)
gacnnnnnntgg
3130
BspCNI(4)
ctcag
2960,3126,3535,2420
BspHI
(1)
tcatga
3090
BspMI(3)
acctgc
1118,1347,1710
BsrBI(5)
ccgctc
191,335,635,2062,3879
BsrDI(3)
gcaatg
2863,1677,2681
BsrFI(4)
rccggy
676,1770,1951,2837
BsrI(20)
actgg
277,1032,2446,3394,3407,81,326,393,419,864
BssHII
(1)
gcgcgc
1848
BssSI(3)
cacgag
2043,2253,3637
BstAPI
(1)
gcannnnntgc
1002
BstF5I(10)
ggatg
463,1632,1059,1242,1304,1761,1786,2469,2756,2937
BstKTI(25)
gatc
257,507,1263,1280,1286,1291,1310,1621,1699,1780
BstNI(6)
ccwgg
142,430,1836,3649,3662,3783
BstUI(12)
cgcg
33,550,574,594,970,1514,1815,1847,2360,2853
BstXI
(2)
ccannnnnntgg
284,326
BstYI(11)
rgatcy
258,1264,1287,1622,1868,2275,2292,3060,3072,3158
BtgI
(1)
ccrygg
1883
BthCI(34)
gcngc
57,273,330,457,530,562,576,598,1002,1255
BtsI(4)
gcagtg
2524,94,1129,2496
Cac8I(24)
gcnngc
61,90,343,476,500,619,633,676,1101,1125
CdiI(7)
catcg
1888,2669,790,1736,1939,2080,2265
ChaI(25)
gatc
257,507,1263,1280,1286,1291,1310,1621,1699,1780
CjeI(7)
ccannnnnngt
3228,3783,243,482,3129,3229,3307
CjePI(4)
ccannnnnnntc
3277,2538,3010,3130
Csp6I(3)
gtac
245,1754,2438
CviAII(14)
catg
226,342,1139,1320,1665,1851,1882,1908,2474,2510
CviJI(78)
rgcy
41,58,122,144,165,217,239,251,271,331
DdeI(4)
ctnag
2420,2960,3126,3535
DpnI(25)
gatc
257,507,1263,1280,1286,1291,1310,1621,1699,1780
DraI(3)
tttaaa
2342,3034,3053
DraIII
(1)
cacnnngtg
782
DrdI(3)
gacnnnnnngtc
828,1478,3708
EaeI(9)
yggccr
42,272,332,396,1357,1531,1922,1949,2529
EagI(3)
cggccg
272,332,1357
EarI(4)
ctcttc
494,1800,2010,2128
EciI(4)
ggcgga
1964,2764,3592,3738
Eco57MI(5)
ctgrag
3283,1576,2008,2235,2832
EcoHI(6)
ccsgg
1056,1453,1613,2384,2735,3431
EcoICRI
(1)
gagctc
252
EcoO109I
(1)
rggnccy
356
EcoRV
(1)
gatatc
315
EsaBC3I(9)
tcga
337,744,1562,1718,1742,1778,1940,2266,3710
FalI
(1)
aagnnnnnctt
1773
FatI(14)
catg
226,342,1139,1320,1665,1851,1882,1908,2474,2510
FauI(6)
cccgc
89,579,1580,32,500,633
FmuI(8)
ggncc
355,498,786,1966,2557,2779,2796,2875
Fnu4HI(34)
gcngc
57,273,330,457,530,562,576,598,1002,1255
FokI(10)
ggatg
463,1632,1059,1242,1304,1761,1786,2469,2756,2937
FspI(3)
tgcgca
528,1551,2697
GdiII(11)
cggccr
42,272,332,396,1357,1922,1949,272,332,1357
HaeI(4)
wggccw
1531,3334,3786,3797
HaeII(6)
rgcgcy
7,622,630,1202,1450,3566
HaeIII(19)
ggcc
41,271,331,355,395,497,786,928,1356,1530
HaeIV
(2)
gaynnnnnrtc
1628,2922
HgaI(4)
gacgc
3130,3708,549,2380
HhaI(30)
gcgc
6,34,99,527,551,564,573,595,621,629
Hin4I(8)
gaynnnnnvtc
257,346,1628,1658,2848,2922,1628,2922
HindIII
(1)
aagctt
240
HinfI(10)
gantc
46,376,828,850,1935,2069,2922,3439,3835,3910
HinP1I(30)
gcgc
6,34,99,527,551,564,573,595,621,629
HpaII(21)
ccgg
163,675,1056,1183,1354,1431,1453,1481,1612,1702
HphI(6)
ggtga
2202,2443,768,2217,2839,3066
Hpy188I(15)
tcnga
255,721,1266,1289,1415,1759,