载体名称:pGEX-6P-3
1 r! Y2 A* V' @. V# A1 F2 K
$ e6 E3 Q8 R* h/ R载体抗性:Amp
; n4 l, w; l. s+ T( [
6 m5 P2 H7 R8 a9 p% b载体大小:4983bp
# ~/ P% a, K+ m7 K* D
1 @3 J) e9 Z' b+ n" F载体宿主:E.coli
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载体序列来源:NCBI
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载体图谱如下:
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载体序列如下:
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+ m+ e% r4 E" j5 k. ZLOCUS CVU78874 4983 bp DNA circular SYN 03-DEC-1996
" k& U9 u% c& S3 dDEFINITION pGEX-6P-3 cloning vector, complete sequence.
9 J: \ q( f; k/ V! Y. `ACCESSION U78874
8 e, [: H2 K5 \5 s& Q. G9 Z
VERSION U78874.1 GI:1699068
$ N P* ~) e. i: }6 j
KEYWORDS .
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SOURCE Cloning vector pGEX-6P-3
3 _2 i z/ A# Y' c1 o
ORGANISM Cloning vector pGEX-6P-3
2 x$ b5 l/ n; x" x( k! Y
other sequences; artificial sequences; vectors.
9 }9 F$ u! x2 f: y
REFERENCE 1 (bases 1 to 4983)
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AUTHORS Smith,D.B. and Johnson,K.S.
+ a- K& }8 _/ o) D TITLE Single-step purification of polypeptides expressed in Escherichia
+ E+ R+ b/ c7 q. u0 h) w b
coli as fusions with glutathione S-transferase
( _7 k3 S# P- P- R8 x+ P3 Y
JOURNAL Gene 67 (1), 31-40 (1988)
4 d; f& U7 G4 i" ~: w9 q PUBMED 3047011
~% Y9 U" C' {1 p
REFERENCE 2 (bases 918 to 938)
; e! m* E5 D; S5 @ AUTHORS Cordingley,M.G., Callahan,P.L., Sardana,V.V., Garsky,V.M. and
. d/ q8 M$ G4 y# q$ k' R' c Colonno,R.J.
4 l* i* Q# N; M9 [- ]
TITLE Substrate requirements of human rhinovirus 3C protease for peptide
2 A" ?+ D! h3 w/ v5 M cleavage in vitro
5 Q# Q# U$ {2 E, H, I& k
JOURNAL J. Biol. Chem. 265 (16), 9062-9065 (1990)
. C3 {. \' k6 E- M& Y: U PUBMED 2160953
8 F" p4 ^; W, c4 q2 uREFERENCE 3 (bases 1 to 4983)
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AUTHORS Dunst,R.W., Bell,P.A. and English,J.D.
4 p7 Z2 A7 }$ }9 L/ S TITLE Pharmacia Biotech PreScission GST Gene Fusion Expression Vector
; n8 l* T- a# o0 x9 o JOURNAL Unpublished
$ R3 q% O* B) U9 _4 j
REFERENCE 4 (bases 1 to 4983)
1 p) k5 i& f5 A AUTHORS Dunst,R.W., Bell,P.A. and English,J.D.
3 K4 ^& \! S% A5 v y) E TITLE Direct Submission
1 @& z7 [9 B4 Z$ R; G& {* z JOURNAL Submitted (20-NOV-1996) Product Center Molecular Biology Reagents,
/ ?. v2 s/ T; m$ B: ^, n0 O
Pharmacia Biotech Inc., 2202 North Bartlett Avenue, Milwaukee, WI
5 l2 [+ _5 T4 |. U 53202-1009, USA
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FEATURES Location/Qualifiers
% M+ C0 u4 h$ s, _ source 1..4983
. } v% V' ?6 s; [# [" R
/organism="Cloning vector pGEX-6P-3"
4 i1 M" _ m0 \. M/ ]1 P
/mol_type="genomic DNA"
' b B( J6 S+ d /db_xref="taxon:54766"
: \' I. X$ T7 N1 x i1 ^ /lab_host="Escherichia coli"
7 m6 l' ]* z: b) I1 V7 q1 Q
/plasmid="pGEX-6P-3"
7 R$ A4 }% o: K7 I! U( _, S -35_signal 183..188
, q: o# u3 n/ d, I' ~0 Q! c) s /note="tac promoter"
7 p- m5 J1 R: t4 p* @1 T -10_signal 205..211
t% h. R6 A9 l4 x' }5 b/ G
/note="tac promoter; for inducible GST gene fusion
' z# [. a4 t' V5 q2 N2 t( n
expression"
! _: t- W& V, Q& P/ P gene 244..995
1 n; T. J+ z! l3 O9 _+ I
/gene="GST"
( ~; ^6 f9 N! K( J+ o9 M7 W N RBS 244
# D% g" k% w. q' z2 Y
/gene="GST"
2 X! T, { d% V! S/ v6 m CDS 258..995
3 _ e% t4 x' u$ ? /gene="GST"
+ I2 J0 {7 V; z. B /codon_start=1
% k9 q, G$ F% g" V" o3 ^% y
/transl_table=11
% Q! @6 G% }$ G /product="glutathione S-transferase"
: X2 x a1 x. I
/protein_id="AAB37352.1"
& V6 |$ ^$ z% x( }& F* m
/db_xref="GI:1699069"
2 G3 g6 Z X# A7 h+ p8 n" i /translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNK
! E" ?- n C4 L7 J4 E! H3 ^% f3 B
KFELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDI
' W" [6 X1 M# B; `
RYGVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALD
3 M2 E7 G/ j0 T! k VVLYMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK
3 @- Y4 H! i/ x, ?
SDLEVLFQGPLGSPNSRVDSSGRIVTD"
6 \0 L0 P1 l O2 V: ^
misc_feature 918..938
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/gene="GST"
) [$ r) [2 d- ~" }9 i9 g# g8 r
/note="prescission protease recognition/cleavage region"
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misc_feature 945..980
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/gene="GST"
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/note="multiple cloning site (MCS); contains unique
+ }% t0 z: H- V/ F
restriction sites for BamH I, EcoR I, Sma I, Sal I, Xho I,
6 s+ G5 B7 w, v( r5 `
and Not I"
" W* [# G0 v+ V8 I# i* r# n -35_signal 1321..1326
& }9 {3 N; J: Z5 u -10_signal 1344..1349
# r5 g' G( ~- t/ M1 _
/note="beta-lactamase promoter"
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CDS 1391..2251
' `2 P" W/ a' @% ~7 h3 J a /note="ampicillin resistance gene"
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/codon_start=1
+ L" Y. e, D1 h7 ~. t6 d7 i3 y /transl_table=11
7 b# W: M, G: b( x /product="beta-lactamase"
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/protein_id="AAB37353.1"
" n" D0 r1 x! f9 d% J /db_xref="GI:1699070"
0 o3 k! @6 O+ z( O' F /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGY
+ c. t3 m# J" \) w0 J* ] IELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVE
! f3 Y# w% P; q) v YSPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRL
# `7 O f( Q$ x2 \4 I4 _: y
DRWEPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPL
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LRSALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIA
8 d+ \- ?, w' Z P* t5 _4 H; Z
EIGASLIKHW"
; E4 t# r4 l) X) ~9 u% m0 X; b misc_feature 2316..3012
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/note="region necessary for plasmid replication"
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rep_origin 3009
6 U: q! [+ G3 e( |$ ?! |$ ^' m
/note="site of plasmid replication initiation"
/ d& n7 ~; F L( @/ @' G7 q: i CDS 3332..4414
0 h) n2 f* K1 M* k0 ~ /note="overproducing allele"
, F) P: r. H. m4 U
/codon_start=1
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/transl_table=11
& z" r! o, U' F( w( q9 D! M1 v6 l6 K /product="lac repressor"
4 x, T4 j( S8 i5 O) _& p3 m6 ~# a /protein_id="AAB37354.1"
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/db_xref="GI:1699071"
0 B; ^+ |- N N1 j4 l) { o& G /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAE
' {9 g/ J1 s* e) t7 z! V. d
LNYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERS
' F% g" Y( N+ w6 L8 [
GVEACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSII
# O' ]/ v! ]( U8 ~9 ~
FSHEDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREG
* [. H! }1 t- s+ o
DWSAMSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDT
# l. v0 P! V4 o/ J
EDSSCYIPPSTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNT
) M1 D/ f' f& f- j; {" ]) G QTASPRALADSLMQLARQVSRLESGQ"
5 ]/ c- Q* g% c" o: ~ORIGIN
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1 acgttatcga ctgcacggtg caccaatgct tctggcgtca ggcagccatc ggaagctgtg
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61 gtatggctgt gcaggtcgta aatcactgca taattcgtgt cgctcaaggc gcactcccgt
: b1 N b% E5 | 121 tctggataat gttttttgcg ccgacatcat aacggttctg gcaaatattc tgaaatgagc
P, T4 z. ]" _ j' M4 u
181 tgttgacaat taatcatcgg ctcgtataat gtgtggaatt gtgagcggat aacaatttca
# W7 p1 x3 q& C; t0 ^2 f- O 241 cacaggaaac agtattcatg tcccctatac taggttattg gaaaattaag ggccttgtgc
5 }, T; q- L. T$ h* j' J. @ b0 v 301 aacccactcg acttcttttg gaatatcttg aagaaaaata tgaagagcat ttgtatgagc
! ~2 A4 f/ c, `! Q
361 gcgatgaagg tgataaatgg cgaaacaaaa agtttgaatt gggtttggag tttcccaatc
, ]! q6 C( }; N; J2 ~) g 421 ttccttatta tattgatggt gatgttaaat taacacagtc tatggccatc atacgttata
. ]. V- z! ]/ E6 ` 481 tagctgacaa gcacaacatg ttgggtggtt gtccaaaaga gcgtgcagag atttcaatgc
8 T8 l" B, H: e8 p7 L
541 ttgaaggagc ggttttggat attagatacg gtgtttcgag aattgcatat agtaaagact
3 b2 I# m' r9 @ 601 ttgaaactct caaagttgat tttcttagca agctacctga aatgctgaaa atgttcgaag
- }1 ?1 ?" ?3 u/ {" b
661 atcgtttatg tcataaaaca tatttaaatg gtgatcatgt aacccatcct gacttcatgt
5 {2 `0 r) d. Z: e% s1 f 721 tgtatgacgc tcttgatgtt gttttataca tggacccaat gtgcctggat gcgttcccaa
. E/ a3 R( O4 G+ Z8 Y+ {/ O
781 aattagtttg ttttaaaaaa cgtattgaag ctatcccaca aattgataag tacttgaaat
% w% b; S/ i6 P4 \5 e9 c% K% f
841 ccagcaagta tatagcatgg cctttgcagg gctggcaagc cacgtttggt ggtggcgacc
# y7 |; @( i% Z" Z 901 atcctccaaa atcggatctg gaagttctgt tccaggggcc cctgggatcc ccgaattccc
9 Y3 L0 O Y. F 961 gggtcgactc gagcggccgc atcgtgactg actgacgatc tgcctcgcgc gtttcggtga
& H+ Q: t6 X6 c# l. W4 t
1021 tgacggtgaa aacctctgac acatgcagct cccggagacg gtcacagctt gtctgtaagc
; A$ I7 k7 h9 ]0 s 1081 ggatgccggg agcagacaag cccgtcaggg cgcgtcagcg ggtgttggcg ggtgtcgggg
- E/ s5 @* A6 s5 ^: F
1141 cgcagccatg acccagtcac gtagcgatag cggagtgtat aattcttgaa gacgaaaggg
4 c+ t, z7 e7 c/ d 1201 cctcgtgata cgcctatttt tataggttaa tgtcatgata ataatggttt cttagacgtc
8 k4 |; ^( s. N- c) \ 1261 aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca
* J, `, s' r2 P/ _( Z! @& B# f 1321 ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa
2 c( g: V. T8 H5 p; q5 _ 1381 aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt ttgcggcatt
8 q" p/ |& \: q7 [0 \9 |( r
1441 ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg ctgaagatca
3 M) m9 `/ n# T4 X6 L, Z$ i2 e
1501 gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga tccttgagag
9 E( [; l# K4 q, o3 s* Y' R
1561 ttttcgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc tatgtggcgc
6 U7 I% P. Z; C% D# z/ W( k 1621 ggtattatcc cgtgttgacg ccgggcaaga gcaactcggt cgccgcatac actattctca
' u* x2 {) I f! f" U% T4 S2 b 1681 gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg gcatgacagt
, e4 t) F0 u' K9 z4 \6 ` 1741 aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca acttacttct
: t' o2 O: ?" A' R5 N! k 1801 gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg gggatcatgt
7 i+ D# \0 n+ X6 S; \
1861 aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg acgagcgtga
! Q. e8 ]: p+ [. Q5 a 1921 caccacgatg cctgcagcaa tggcaacaac gttgcgcaaa ctattaactg gcgaactact
* i3 b' c4 P6 T7 |
1981 tactctagct tcccggcaac aattaataga ctggatggag gcggataaag ttgcaggacc
U- R3 \* u. \& n) z( D 2041 acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg gagccggtga
# ]7 Z: |% j/ v0 d: z9 C8 N
2101 gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct cccgtatcgt
" t3 j# H+ z4 r
2161 agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac agatcgctga
! w: { t& o. V
2221 gataggtgcc tcactgatta agcattggta actgtcagac caagtttact catatatact
' R6 K- ~) x" O
2281 ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga
1 _0 O2 j! L( K
2341 taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt
3 ~' u! Y" C/ {) E 2401 agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca
, s; \' Z4 S! {' Q: |! A$ u7 j. ~4 n 2461 aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct
4 @2 `3 f( n; |& y 2521 ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgtcc ttctagtgta
/ d# _3 r; K+ d0 k 2581 gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct
( f4 x6 Z6 F+ i+ H3 R4 D
2641 aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc
8 u0 q8 z# c3 p5 ^: F 2701 aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca
: [) y( c6 a) H; L }" g 2761 gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga
: ^0 w. y8 t2 `8 _. s
2821 aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg
: F5 l# N" e+ o# x s9 u
2881 aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt
: ~3 }' I. O& l: _7 t
2941 cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag
& e, }; f' t" ^* K$ ]7 y! G/ i
3001 cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt
3 L4 T4 \ l) Z+ r
3061 tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgcctt
6 q" C, o3 h5 [4 Z$ |! f
3121 tgagtgagct gataccgctc gccgcagccg aacgaccgag cgcagcgagt cagtgagcga
@; d3 `" Z7 c; U7 ~% ]( s2 r) v 3181 ggaagcggaa gagcgcctga tgcggtattt tctccttacg catctgtgcg gtatttcaca
0 ?" F* n5 t: c$ \8 p 3241 ccgcataaat tccgacacca tcgaatggtg caaaaccttt cgcggtatgg catgatagcg
& l5 A+ i( I7 l- u3 |8 b5 e! W 3301 cccggaagag agtcaattca gggtggtgaa tgtgaaacca gtaacgttat acgatgtcgc
" @3 \6 g" u/ M/ m
3361 agagtatgcc ggtgtctctt atcagaccgt ttcccgcgtg gtgaaccagg ccagccacgt
+ d( F' [' q" i+ d: w2 g 3421 ttctgcgaaa acgcgggaaa aagtggaagc ggcgatggcg gagctgaatt acattcccaa
i$ s8 m" I o7 y; D 3481 ccgcgtggca caacaactgg cgggcaaaca gtcgttgctg attggcgttg ccacctccag
2 i2 |8 c" h4 L$ C0 S. G& o 3541 tctggccctg cacgcgccgt cgcaaattgt cgcggcgatt aaatctcgcg ccgatcaact
4 A# A6 Q6 |# k& ]. I3 t' f 3601 gggtgccagc gtggtggtgt cgatggtaga acgaagcggc gtcgaagcct gtaaagcggc
7 v6 W9 a3 e3 G" D# G# @4 N
3661 ggtgcacaat cttctcgcgc aacgcgtcag tgggctgatc attaactatc cgctggatga
9 b. f1 G0 m, q: }
3721 ccaggatgcc attgctgtgg aagctgcctg cactaatgtt ccggcgttat ttcttgatgt
/ ~/ r3 j6 {5 C- b! _ 3781 ctctgaccag acacccatca acagtattat tttctcccat gaagacggta cgcgactggg
- r7 Z! Q2 M- _* j$ @
3841 cgtggagcat ctggtcgcat tgggtcacca gcaaatcgcg ctgttagcgg gcccattaag
. \# w. N- C$ S$ t
3901 ttctgtctcg gcgcgtctgc gtctggctgg ctggcataaa tatctcactc gcaatcaaat
7 N. r1 i7 G1 r; V
3961 tcagccgata gcggaacggg aaggcgactg gagtgccatg tccggttttc aacaaaccat
( ~1 T S9 o3 e7 E0 u5 k/ @* N6 E
4021 gcaaatgctg aatgagggca tcgttcccac tgcgatgctg gttgccaacg atcagatggc
7 b/ m2 v* D( `7 B+ x- i 4081 gctgggcgca atgcgcgcca ttaccgagtc cgggctgcgc gttggtgcgg atatctcggt
5 J6 {9 u( V6 e
4141 agtgggatac gacgataccg aagacagctc atgttatatc ccgccgtcaa ccaccatcaa
( k5 D" m- Q1 @9 I. r
4201 acaggatttt cgcctgctgg ggcaaaccag cgtggaccgc ttgctgcaac tctctcaggg
; Z0 |( o( x4 g/ v6 m* H- Y2 P# a
4261 ccaggcggtg aagggcaatc agctgttgcc cgtctcactg gtgaaaagaa aaaccaccct
5 p& L+ K/ {$ j" u$ T; K* ^) o% N 4321 ggcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc
3 { Y8 ]% m2 }# m# {
4381 acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc
v8 m3 X7 J8 o& E; K6 V- | {
4441 tcactcatta ggcaccccag gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa
9 x3 v; c3 `3 H! M; K& J 4501 ttgtgagcgg ataacaattt cacacaggaa acagctatga ccatgattac ggattcactg
2 m, J' p7 H$ V: x, J' R% Q$ v
4561 gccgtcgttt tacaacgtcg tgactgggaa aaccctggcg ttacccaact taatcgcctt
7 S6 v, ]' i& I. f' u 4621 gcagcacatc cccctttcgc cagctggcgt aatagcgaag aggcccgcac cgatcgccct
8 t- n9 U8 }0 V* v 4681 tcccaacagt tgcgcagcct gaatggcgaa tggcgctttg cctggtttcc ggcaccagaa
w) N/ I! @, k/ C/ y0 Y u( G5 T6 R 4741 gcggtgccgg aaagctggct ggagtgcgat cttcctgagg ccgatactgt cgtcgtcccc
/ Y' }& W5 v" E1 V! u7 R
4801 tcaaactggc agatgcacgg ttacgatgcg cccatctaca ccaacgtaac ctatcccatt
( i% U, F: n0 q) K( t5 }
4861 acggtcaatc cgccgtttgt tcccacggag aatccgacgg gttgttactc gctcacattt
8 w* o. K/ |/ i0 i 4921 aatgttgatg aaagctggct acaggaaggc cagacgcgaa ttatttttga tggcgttgga
1 w- Q% q# u7 Q: }$ T6 B
4981 att
0 M; s9 X2 ?) |5 [/ {/ e* V) E
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本帖最后由 wcg129 于 08-1-1 22:16 编辑 ]