Ejercicio 1

Disea los oligonucletidos necesarios para amplificar el gen de la enzima Dcarbamilasa de Agrobacterium de forma que se exprese con una cola de 6 histidinas
en el extremo amino terminal. No es necesario tener en cuenta la pauta de lectura ya
que el plsmido est diseado para que no haya fusin con ningn pptido.
X91070. Agrobacterium radiobacter
DEFINITION A.radiobacter D-hydantoinase gene, and D-N-alpha-carbamylase gene.
KEYWORDS D-hydantoinase gene; D-N-alpha-carbamilase gene.
SOURCE
Agrobacterium tumefaciens (Rhizobium radiobacter)
ORGANISM
Agrobacterium
tumefaciens
Bacteria;
Proteobacteria;
Alphaproteobacteria; Rhizobiales; Rhizobiaceae; Rhizobium/Agrobacterium group;
Agrobacterium.
gene
1618..2532 /gene="D-N-alpha-carbamilase"
CDS
1618..2532 /gene="D-N-alpha-carbamilase"
/translation="MTRQMILAVGQQGPIARAETREQVVGRLLDMLTNAASRGVNFIVFPELA
LTTFFPRWHFTDEAELDSFYETEMPGPVVRPLFETAAELGIGFNLGYAELVVEGGVKR
RFNTSILVDKSGKIVGKYRKIHLPGHKEYEAYRPFQHLEKRYFEPGDLGFPVYDVDAA
KMGMFICNDRRWPETWRVMGLKGAEIICGGYNTPTHNPPVPQHDHLTSFHHLLSMQ
AGSYQNGAWSAAAGKVGMEEGCMLLGHSCIVAPTGEIVALTTTLEDEVITAAVDLDR
CRELREHIFNFKAHRQPQHYGLIAEF"
1021 ggatcgagca cgatgatatg gtagccgtag tcgatcgccg acttgccgcc ggccatgccg
1081 tcccatttgg cgaccgcctc cctcaggcta tggccgcggt cctgctggca gaaatcgacg
1141 atggtcgtcg tgccgccaca ggcggccgcg accgtcgcgg ttgcgaatgt gtcggccgac
1201 tgcgtgttga agctgaccgt ctcgacatgc gtatgaacgt cgatgccgcc cggaaaaacg
1261 tagcggccgg aggcgtcgat tgtccggccg gccgggccga acgttccgcc gatctgggcg
1321 atcttgccat ccttgattcc gagatcggcg ggagaaatcc cgtccgcggt tacgatggtt
1381 ccgttcttga tgatgatatc cataaagcag ctctcagggt tgatggataa attctatatg
1441 cggtatgatg ttctttatat aaagttttca tgttgccttg tatctgtcaa gcgggaaggg
1501 aagttctccg gaatcggcgc tgcgagggaa cgtatcgagt ttcgattaga cgcggttgaa
1561 agcgcagcga gtcattgaac tacggaacct ctgccaaacc ctattcggcg agctggaatg
1621 acacgtcaga tgatacttgc tgtcggacag caaggcccca tcgcgcgagc ggagacacgc
1681 gaacaggtgg ttggccgcct cctcgacatg ttgacgaacg cagccagccg gggcgtgaac
1741 ttcatcgtct ttcccgagct tgcgctcacg accttcttcc cgcgctggca tttcaccgac
1801 gaggccgagc tcgatagctt ctatgagacc gaaatgcccg gcccggtggt ccgtccactc
1861 tttgagacgg ccgccgaact cgggatcggc ttcaatctgg gctacgccga actcgtcgtc
1921 gaaggcggcg tcaagcgtcg cttcaacacg tccattctgg tggataagtc aggcaagatc
1981 gtcggcaagt atcgtaagat ccatttgccg ggtcacaagg agtacgaggc ctaccggccg
2041 ttccagcatc ttgaaaagcg ttatttcgag ccgggcgatc tcggcttccc ggtctatgac
2101 gtcgacgccg cgaaaatggg gatgttcatc tgcaacgatc gccgctggcc tgaaacgtgg
2161 cgggtgatgg gacttaaggg cgccgagatc atctgcggcg gctacaacac gccgacccac
2221 aatccccccg ttccccagca cgaccatctg acgtccttcc accaccttct gtcgatgcag
.. 2281 gccgggtcgt accaaaacgg cgcctggtcc gcggcggccg gcaaggtcgg catggaggag
2341 gggtgcatgc tgctcggcca ttcgtgcatc gtggcgccga ccggcgaaat cgttgccctg
2401 accacgacgt tggaagacga ggtgatcacc gccgccgtcg atctcgaccg ctgccgggaa
2461 ctgcgcgaac acatcttcaa tttcaaagcc catcgtcagc cacagcacta cggtctgatc
2521 gcggaattct ga
Enzimas que no cortan el fragmento completo:
AarI
BamHI
BsrGI
ClaI
DraI
DraIII
Eco57I
FspI
HindIII
KpnI
MluI
MscI
MunI
NdeI
NheI
OliI
PacI
PstI
PvuII
RsrII
SanDI
SbfI
ScaI
SexAI
SgfI
SgrAI
SmaI
SnaBI
SpeI
SrfI
SspI
SwaI
TatI
XbaI
XhoI
XmaI

NdeI
EcoRI
KpnI
SmaI
BamHI
XbaI
PstI
HindIII

PROMOTOR

XmnI

SITIO DE CLONACION

ScaI
2500

2000

500

pBBM1
2686 bps
1000
1500