"
His+FluA+SplitLinker+Foki
1 gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt
caccgtgaaa agccccttta cacgcgcctt ggggataaac aaataaaaag atttatgtaa
r w h f s g k c a r n p y l f i f l n t
61 caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa
gtttatacat aggcgagtac tctgttattg ggactattta cgaagttatt ataacttttt
f k y v s a h e t i t l i n a s i i l k
121 ggaagagtat gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt
ccttctcata ctcataagtt gtaaaggcac agcgggaata agggaaaaaa cgccgtaaaa
k e e y e y s t f p c r p y s l f c g i
>>........................AmpR..........................>
m s i q h f r v a l i p f f a a f
181 gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt
cggaaggaca aaaacgagtg ggtctttgcg accactttca ttttctacga cttctagtca
l p s c f c s p r n a g e s k r c - r s
>.............................AmpR..............................>
c l p v f a h p e t l v k v k d a e d q
241 tgggtgcacg agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt
acccacgtgc tcacccaatg tagcttgacc tagagttgtc gccattctag gaactctcaa
v g c t s g l h r t g s q q r - d p - e
>.............................AmpR..............................>
l g a r v g y i e l d l n s g k i l e s
301 ttcgccccga agaacgtttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg
aagcggggct tcttgcaaaa ggttactact cgtgaaaatt tcaagacgat acaccgcgcc
f s p r r t f s n d e h f - s s a m w r
>.............................AmpR..............................>
f r p e e r f p m m s t f k v l l c g a
361 tattatcccg tattgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga
ataatagggc ataactgcgg cccgttctcg ttgagccagc ggcgtatgtg ataagagtct
g i i p y - r r a r a t r s p h t l f s
>.............................AmpR..............................>
v l s r i d a g q e q l g r r i h y s q
421 atgacttggt tgagtactca ccagtcacag aaaagcatct tacggatggc atgacagtaa
tactgaacca actcatgagt ggtcagtgtc ttttcgtaga atgcctaccg tactgtcatt
e - l g - v l t s h r k a s y g w h d s
>.............................AmpR..............................>
n d l v e y s p v t e k h l t d g m t v
481 gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga
ctcttaatac gtcacgacgg tattggtact cactattgtg acgccggttg aatgaagact
k r i m q c c h n h e - - h c g q l t s
>.............................AmpR..............................>
r e l c s a a i t m s d n t a a n l l l
541 caacgatcgg aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa
gttgctagcc tcctggcttc ctcgattggc gaaaaaacgt gttgtacccc ctagtacatt
d n d r r t e g a n r f f a q h g g s c
>.............................AmpR..............................>
t t i g g p k e l t a f l h n m g d h v
601 ctcgccttga tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca
gagcggaact agcaaccctt ggcctcgact tacttcggta tggtttgctg ctcgcactgt
n s p - s l g t g a e - s h t k r r a -
>.............................AmpR..............................>
t r l d r w e p e l n e a i p n d e r d
661 ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact attaactggc gaactactta
ggtgctacgg acatcgttac cgttgttgca acgcgtttga taattgaccg cttgatgaat
h h d a c s n g n n v a q t i n w r t t
>.............................AmpR..............................>
t t m p v a m a t t l r k l l t g e l l
721 ctctagcttc ccggcaacaa ttaatagact ggatggaggc ggataaagtt gcaggaccac
gagatcgaag ggccgttgtt aattatctga cctacctccg cctatttcaa cgtcctggtg
y s s f p a t i n r l d g g g - s c r t
>.............................AmpR..............................>
t l a s r q q l i d w m e a d k v a g p
781 ttctgcgctc ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc
aagacgcgag ccgggaaggc cgaccgacca aataacgact atttagacct cggccactcg
t s a l g p s g w l v y c - - i w s r -
>.............................AmpR..............................>
l l r s a l p a g w f i a d k s g a g e
841 gtggttctcg cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag
caccaagagc gccatagtaa cgtcgtgacc ccggtctacc attcgggagg gcatagcatc
a w f s r y h c s t g a r w - a l p y r
>.............................AmpR..............................>
r g s r g i i a a l g p d g k p s r i v
901 ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga
aatagatgtg ctgcccctca gtccgttgat acctacttgc tttatctgtc tagcgactct
s y l h d g e s g n y g - t k - t d r -
>.............................AmpR..............................>
v i y t t g s q a t m d e r n r q i a e
961 taggtgcctc actgattaag cattggtaac tgtcagacca agtttactca tatatacttt
atccacggag tgactaattc gtaaccattg acagtctggt tcaaatgagt atatatgaaa
d r c l t d - a l v t v r p s l l i y t
>............AmpR............>>
i g a s l i k h w -
1021 agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata
tctaactaaa ttttgaagta aaaattaaat tttcctagat ccacttctag gaaaaactat
l d - f k t s f l i - k d l g e d p f -
1081 atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag
tagagtactg gttttaggga attgcactca aaagcaaggt gactcgcagt ctggggcatc
- s h d q n p l t - v f v p l s v r p r
1141 aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa
ttttctagtt tcctagaaga actctaggaa aaaaagacgc gcattagacg acgaacgttt
r k d q r i f l r s f f s a r n l l l a
1201 caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt
gtttttttgg tggcgatggt cgccaccaaa caaacggcct agttctcgat ggttgagaaa
n k k t t a t s g g l f a g s r a t n s
1261 ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc
aaggcttcca ttgaccgaag tcgtctcgcg tctatggttt atgacaagaa gatcacatcg
f s e g n w l q q s a d t k y c s s s v
1321 cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa
gcatcaatcc ggtggtgaag ttcttgagac atcgtggcgg atgtatggag cgagacgatt
a v v r p p l q e l c s t a y i p r s a
1381 tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa
aggacaatgg tcaccgacga cggtcaccgc tattcagcac agaatggccc aacctgagtt
n p v t s g c c q w r - v v s y r v g l
1441 gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc
ctgctatcaa tggcctattc cgcgtcgcca gcccgacttg ccccccaagc acgtgtgtcg
k t i v t g - g a a v g l n g g f v h t
1501 ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa
ggtcgaacct cgcttgctgg atgtggcttg actctatgga tgtcgcactc gatactcttt
a q l g a n d l h r t e i p t a - a m r
1561 gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa
cgcggtgcga agggcttccc tctttccgcc tgtccatagg ccattcgccg tcccagcctt
k r h a s r r e k g g q v s g k r q g r
1621 caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg
gtcctctcgc gtgctccctc gaaggtcccc ctttgcggac catagaaata tcaggacagc
n r r a h e g a s r g k r l v s l - s c
1681 ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc
ccaaagcggt ggagactgaa ctcgcagcta aaaacactac gagcagtccc cccgcctcgg
r v s p p l t - a s i f v m l v r g a e
1741 tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg
ataccttttt gcggtcgttg cgccggaaaa atgccaagga ccggaaaacg accggaaaac
p m e k r q q r g l f t v p g l l l a f
1801 ctcattaggc accccaggct ttacccgaac gaccgagcgc agcgagtcag tgagcgagga
gagtaatccg tggggtccga aatgggcttg ctggctcgcg tcgctcagtc actcgctcct
c s l g t p g f t r t t e r s e s v s e
1861 agcggagagc gcccaatacg caaggaaaca gctatgacca tgttaatgca gctggcacga
tcgcctctcg cgggttatgc gttcctttgt cgatactggt acaattacgt cgaccgtgct
e a e s a q y a r k q l - p c - c s w h
1921 caggtttccc gactggaaag cgggcagtga gcggaaggcc catgaggcca gttaattaag
gtccaaaggg ctgacctttc gcccgtcact cgccttccgg gtactccggt caattaattc
d r f p d w k a g s e r k a h e a s - l
NotI
--+------
EcoRI XbaI NgoMIV
-+----- -+---- -+----
1981 aggtaccgaa ttcgcggccg cttctagatg gccggccatc atcatcatca tcataccggc
tccatggctt aagcgccggc gaagatctac cggccggtag tagtagtagt agtatggccg
r g t e f a a a s r w p a i i i i i i p
>>.............ORF-1...............>
m a g h h h h h h t g
>>....His Tag.....>>
h h h h h h
2041 gacgtgtacc acgacggcgc ctgccccgaa gtgaagcccg tggacaactt cgactggtcc
ctgcacatgg tgctgccgcg gacggggctt cacttcgggc acctgttgaa gctgaccagg
a t c t t t a p a p k - s p w t t s t g
>.............................ORF-1.............................>
d v y h d g a c p e v k p v d n f d w s
>>............................FluA..............................>
d v y h d g a c p e v k p v d n f d w s
2101 cagtaccacg gcaagtggtg ggaggtggcc aagtacccca gccccaacgg caagtatggc
gtcatggtgc cgttcaccac cctccaccgg ttcatggggt cggggttgcc gttcataccg
p s t t a s g g r w p s t p a p t a s m
>.............................ORF-1.............................>
q y h g k w w e v a k y p s p n g k y g
>.............................FluA..............................>
q y h g k w w e v a k y p s p n g k y g
2161 aagtgcggct gggccgagta cacccccgag ggcaagagcg tgaaggtgtc cagatacgac
ttcacgccga cccggctcat gtgggggctc ccgttctcgc acttccacag gtctatgctg
a s a a g p s t p p r a r a - r c p d t
>.............................ORF-1.............................>
k c g w a e y t p e g k s v k v s r y d
>.............................FluA..............................>
k c g w a e y t p e g k s v k v s r y d
2221 gtgatccacg gcaaagaata cttcatggaa ggcaccgcct accccgtggg cgacagcaag
cactaggtgc cgtttcttat gaagtacctt ccgtggcgga tggggcaccc gctgtcgttc
t - s t a k n t s w k a p p t p w a t a
>.............................ORF-1.............................>
v i h g k e y f m e g t a y p v g d s k
>.............................FluA..............................>
v i h g k e y f m e g t a y p v g d s k
2281 atcggcaaga tctaccacag ccggaccgtg ggcggctaca ccagaaagac cgtgttcaac
tagccgttct agatggtgtc ggcctggcac ccgccgatgt ggtctttctg gcacaagttg
r s a r s t t a g p w a a t p e r p c s
>.............................ORF-1.............................>
i g k i y h s r t v g g y t r k t v f n
>.............................FluA..............................>
i g k i y h s r t v g g y t r k t v f n
2341 gtgctgtcca ccgacaacaa gaactacatc atcggctaca gctgccgcta cgacgaggac
cacgacaggt ggctgttgtt cttgatgtag tagccgatgt cgacggcgat gctgctcctg
t c c p p t t r t t s s a t a a a t t r
>.............................ORF-1.............................>
v l s t d n k n y i i g y s c r y d e d
>.............................FluA..............................>
v l s t d n k n y i i g y s c r y d e d
2401 aagaagggcc actgggacca cgtgtgggtg ctgtcccggt ccatggtgct gaccggcgag
ttcttcccgg tgaccctggt gcacacccac gacagggcca ggtaccacga ctggccgctc
t r r a t g t t c g c c p g p w c - p a
>.............................ORF-1.............................>
k k g h w d h v w v l s r s m v l t g e
>.............................FluA..............................>
k k g h w d h v w v l s r s m v l t g e
2461 gccaagaccg ccgtggagaa ctacctgatc ggcagccccg tggtggacag ccagaaactg
cggttctggc ggcacctctt gatggactag ccgtcggggc accacctgtc ggtctttgac
r p r p p w r t t - s a a p w w t a r n
>.............................ORF-1.............................>
a k t a v e n y l i g s p v v d s q k l
>.............................FluA..............................>
a k t a v e n y l i g s p v v d s q k l
2521 gtgtacagcg acttcagcga ggccgcctgc aaagtgaaca acaccggccg accagcctgt
cacatgtcgc tgaagtcgct ccggcggacg tttcacttgt tgtggccggc tggtcggaca
w c t a t s a r p p a k - t t p a d q p
>.............................ORF-1.............................>
v y s d f s e a a c k v n n t g r p a c
>....................FluA...................>>
v y s d f s e a a c k v n n
Split-Linker >>..........>
r p a c
2581 aagattccaa atgacctgaa gcagaaagtt atgaatcaca ccggcaaatc tgaactggag
ttctaaggtt tactggactt cgtctttcaa tacttagtgt ggccgtttag acttgacctc
v r f q m t - s r k l - i t p a n l n w
>.............................ORF-1.............................>
k i p n d l k q k v m n h t g k s e l e
>..............Split-Linker.............>>
k i p n d l k q k v m n h
>>.'Fok inaktiv'..>
g k s e l e
2641 gagaaaaaat ccgagctgcg ccacaaactg aaatatgtgc ctcacgagta tatcgaactg
ctctttttta ggctcgacgc ggtgtttgac tttatacacg gagtgctcat atagcttgac
r r k n p s c a t n - n m c l t s i s n
>.............................ORF-1.............................>
e k k s e l r h k l k y v p h e y i e l
>.........................'Fok inaktiv'.........................>
e k k s e l r h k l k y v p h e y i e l
2701 atcgagatcg cccgtaatag tacccaagac cgtatcctgg aaatgaaagt gatggagttc
tagctctagc gggcattatc atgggttctg gcataggacc tttactttca ctacctcaag
- s r s p v i v p k t v s w k - k - w s
>.............................ORF-1.............................>
i e i a r n s t q d r i l e m k v m e f
>.........................'Fok inaktiv'.........................>
i e i a r n s t q d r i l e m k v m e f
2761 ttcatgaaag tctatggcta tcgtggcaaa catctgggtg gtagccgtaa accagcaggt
aagtactttc agataccgat agcaccgttt gtagacccac catcggcatt tggtcgtcca
s s - k s m a i v a n i w v v a v n q q
>.............................ORF-1.............................>
f m k v y g y r g k h l g g s r k p a g
>.........................'Fok inaktiv'.........................>
f m k v y g y r g k h l g g s r k p a g
2821 gccatttata ccgttggttc cccgatcgat tatggcgtta tcgtggccac aaaagcgtat
cggtaaatat ggcaaccaag gggctagcta ataccgcaat agcaccggtg ttttcgcata
v p f i p l v p r s i m a l s w p q k r
>.............................ORF-1.............................>
a i y t v g s p i d y g v i v a t k a y
>.........................'Fok inaktiv'.........................>
a i y t v g s p i d y g v i v a t k a y
2881 tctggcggtt ataatctgcc gattggtcag gctgatgaga tggaacgtta tgtggaagag
agaccgccaa tattagacgg ctaaccagtc cgactactct accttgcaat acaccttctc
i l a v i i c r l v r l m r w n v m w k
>.............................ORF-1.............................>
s g g y n l p i g q a d e m e r y v e e
>.........................'Fok inaktiv'.........................>
s g g y n l p i g q a d e m e r y v e e
2941 aatcagaccc gtaacaaaca tctgaacccg aacgaatggt ggaaagtgta tccgtcaagt
ttagtctggg cattgtttgt agacttgggc ttgcttacca cctttcacat aggcagttca
r i r p v t n i - t r t n g g k c i r q
>.............................ORF-1.............................>
n q t r n k h l n p n e w w k v y p s s
>.........................'Fok inaktiv'.........................>
n q t r n k h l n p n e w w k v y p s s
3001 gtcaccgagt tcaaatttct gttcgtgagc ggccacttta aaggcaacta taaagcccag
cagtggctca agtttaaaga caagcactcg ccggtgaaat ttccgttgat atttcgggtc
v s p s s n f c s - a a t l k a t i k p
>.............................ORF-1.............................>
v t e f k f l f v s g h f k g n y k a q
>.........................'Fok inaktiv'.........................>
v t e f k f l f v s g h f k g n y k a q
3061 ctgactcgtc tgaaccatat caccaatagc aatggggcag tgctgagtgt tgaggaactg
gactgagcag acttggtata gtggttatcg ttaccccgtc acgactcaca actccttgac
s - l v - t i s p i a m g q c - v l r n
>.............................ORF-1.............................>
l t r l n h i t n s n g a v l s v e e l
>.........................'Fok inaktiv'.........................>
l t r l n h i t n s n g a v l s v e e l
3121 ctgatcggtg gagaaatgat caaagcaggc accctgactc tggaagaagt tcgccgtaaa
gactagccac ctctttacta gtttcgtccg tgggactgag accttcttca agcggcattt
c - s v e k - s k q a p - l w k k f a v
>.............................ORF-1.............................>
l i g g e m i k a g t l t l e e v r r k
>.........................'Fok inaktiv'.........................>
l i g g e m i k a g t l t l e e v r r k
PstI
------+
SpeI NotI
-+---- --+-----
3181 ttcaacaatg gcgagatcaa ttttaccggt taatactagt agcggccgct gcagagctca
aagttgttac cgctctagtt aaaatggcca attatgatca tcgccggcga cgtctcgagt
n s t m a r s i l p v n t s s g r c r a
>..............ORF-1..............>>
f n n g e i n f t g -
>..........'Fok inaktiv'.........>>
f n n g e i n f t g
3241 tggcgcgcct aggccttgac ggccttccgc caattcgccc tatagtgagt cgtattacgt
accgcgcgga tccggaactg ccggaaggcg gttaagcggg atatcactca gcataatgca
h g a p r p - r p s a n s p y s e s y y
3301 cgcgctcact ggccgtcgtt ttacaacgtc gtgactggga aaaccctggc gttacccaac
gcgcgagtga ccggcagcaa aatgttgcag cactgaccct tttgggaccg caatgggttg
v a l t g r r f t t s - l g k p w r y p
3361 ttaatcgcct tgcagcacat ccccctttcg ccagctggcg taatagcgaa gaggcccgca
aattagcgga acgtcgtgta gggggaaagc ggtcgaccgc attatcgctt ctccgggcgt
t - s p c s t s p f r q l a - - r r g p
3421 ccgaaacgcc cttcccaaca gttgcgcagc ctgaatggcg aatgggagcg ccctgtagcg
ggctttgcgg gaagggttgt caacgcgtcg gacttaccgc ttaccctcgc gggacatcgc
h r n a l p n s c a a - m a n g s a l -
3481 gccactcaac cctatctcgg tctattcttt tgatttataa gggattttgc cgatttcggc
cggtgagttg ggatagagcc agataagaaa actaaatatt ccctaaaacg gctaaagccg
r p l n p i s v y s f d l - g i l p i s
3541 ctattggtta aaaaatgagc tgatttaaca aaaatttaac gcgaatttta acaaaatatt
gataaccaat tttttactcg actaaattgt ttttaaattg cgcttaaaat tgttttataa
a y w l k n e l i - q k f n a n f n k i
3601 aacgcttaca atttag
ttgcgaatgt taaatc
l t l t i -
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