Team:UC-Santa Cruz/Project/Pumps

From 2013.igem.org

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<div class=Section1>
<div class=Section1>
 +
 +
<h3>
 +
Ion Pumps
 +
</h3>
<p class=MsoNormal><span style='font-family:"Times New Roman";mso-fareast-font-family:
<p class=MsoNormal><span style='font-family:"Times New Roman";mso-fareast-font-family:
-
"Times New Roman"'>The goal of the ion pumps group was to select and obtain ion
+
"Times New Roman"'>The goal of the ion pumps group was to find and obtain light driven sodium
-
pumps to be expressed in <span class=SpellE>C.Cresentus</span>. <span
+
and chloride pumps to be expressed in <span class=SpellE>C. cresentus</span>. <span
-
class=GramE>Selection of ion pumps were</span> based on ion selectivity and
+
class=GramE>Choice of ion pumps was</span> based on ion selectivity and
structure simplicity.<o:p></o:p></span></p>
structure simplicity.<o:p></o:p></span></p>
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<p class=MsoNormal><span style='font-family:"Times New Roman";mso-fareast-font-family:
<p class=MsoNormal><span style='font-family:"Times New Roman";mso-fareast-font-family:
"Times New Roman"'>Two pumps were used for this project. The first choice was
"Times New Roman"'>Two pumps were used for this project. The first choice was
-
the widely studied ion pump, <span class=SpellE>Halorhodopsin</span> (HR), an
+
the widely studied <span class=SpellE>Halorhodopsin</span> (HR), an
-
inward pointing conductor. For the sodium pump we chose the newly discovered
+
inward pointing chloride conductor. For the sodium pump we chose the newly discovered
-
KR2, an outward pointing conductor.<o:p></o:p></span></p>
+
KR2, an outward facing pump.<o:p></o:p></span></p>
<p class=MsoNormal><span style='font-family:"Times New Roman";mso-fareast-font-family:
<p class=MsoNormal><span style='font-family:"Times New Roman";mso-fareast-font-family:
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<p class=MsoNormal><span style='font-family:"Times New Roman";mso-fareast-font-family:
<p class=MsoNormal><span style='font-family:"Times New Roman";mso-fareast-font-family:
"Times New Roman"'>First, we searched the <span class=SpellE>iGEM</span>
"Times New Roman"'>First, we searched the <span class=SpellE>iGEM</span>
-
catalog for parts for anything containing the coding sequence for HR. We found
+
catalog for any parts containing the coding sequence for HR. We found
three parts<span class=GramE>;</span> BBaK559000, BBaK9001 and BBaK559010 and
three parts<span class=GramE>;</span> BBaK559000, BBaK9001 and BBaK559010 and
-
ordered them. Upon <span class=SpellE>desiging</span> sequencing primers and
+
requested them for use in our project. After isolating and
-
sequencing of these plasmids, we found them to containing incomplete versions
+
sequencing these plasmids, we found them to contain incomplete versions
-
of the coding sequence for HR from <span class=SpellE>N.pharaonis</span>.
+
of the coding sequence of Halorhodopsin from <span class=SpellE>N.pharaonis</span>.
Another problem was that the sequence of the HR in the parts contained an extra
Another problem was that the sequence of the HR in the parts contained an extra
<span class=GramE>base which</span> was not in agreement with the coding
<span class=GramE>base which</span> was not in agreement with the coding
-
sequence the team who made the part reported. Luckily, running in parallel to
+
sequence the team who made the part reported. With this we searched for a new
-
analysis of the parts we had another way to obtain the DNA coding for HR.<span
+
source to obtain the Halorhodopsin gene from.<span
style="mso-spacerun: yes">&nbsp;&nbsp; </span><o:p></o:p></span></p>
style="mso-spacerun: yes">&nbsp;&nbsp; </span><o:p></o:p></span></p>
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<p class=MsoNormal><span style='font-family:"Times New Roman";mso-fareast-font-family:
<p class=MsoNormal><span style='font-family:"Times New Roman";mso-fareast-font-family:
-
"Times New Roman"'>The other attempt was a search for every organism that
+
"Times New Roman"'>The next attempt was to search other organisms that
-
contained the gene was performed. Soon after, we searched our campus for stocks
+
contained the HR gene. We found that one of these organisms<span
-
of these organisms. It was soon discovered that one of these organisms<span
+
class=GramE>,<span style="mso-spacerun: yes">&nbsp; </span><span class=SpellE>Halobacterium</span></span>
class=GramE>,<span style="mso-spacerun: yes">&nbsp; </span><span class=SpellE>Halobacterium</span></span>
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<span class=SpellE>Salinarium</span> (HS)<span style="mso-spacerun: yes">&nbsp;
+
<span class=SpellE>Salinarum</span> (HS)<span style="mso-spacerun: yes">&nbsp;
-
</span>was in a frozen stock close by our lab on campus. Fortunately, it was
+
</span>was in a frozen stock close by our lab on campus. Fortunately the <span class=SpellE>codon</span> usage of the wild-type HR for H. salinarum was very similar that of C. <span class=SpellE>Cresentus</span>,
-
discovered that the <span class=SpellE>codon</span> usage of the wild-type HR
+
-
for this HS was very much alike that of C. <span class=SpellE>Cresentus</span>,
+
therefore <span class=SpellE>codon</span> optimization was not required.
therefore <span class=SpellE>codon</span> optimization was not required.
Specific primers were then designed to amplify the HR gene out of the HS
Specific primers were then designed to amplify the HR gene out of the HS
-
genome. The primers contain cut sites in order for ligation into our plasmids.
+
genome. The primers contain restriction sites for ligation into our plasmids.
(<span class=GramE>see</span> tags section)<o:p></o:p></span></p>
(<span class=GramE>see</span> tags section)<o:p></o:p></span></p>
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<p class=MsoNormal><span style='font-family:"Times New Roman";mso-fareast-font-family:
<p class=MsoNormal><span style='font-family:"Times New Roman";mso-fareast-font-family:
-
"Times New Roman"'>Since KR2 is a newly discovered Sodium pump in a lab in
+
"Times New Roman"'>The second pump used in this project was a newly discovered light driven sodium pump KR2, from a lab in
-
South Korea, it appeared to be very difficult to acquire the DNA for this
+
South Korea. The organism which</span> contains the KR2 gene, Krokinobacter <span class=SpellE><span
-
particular pump. The organism, Krokinobacter <span class=SpellE><span
+
class=GramE>Eikastus</span></span><span class=GramE>, proved quite difficult to obtain. Additionally, the <span
-
class=GramE>Eikastus</span></span><span class=GramE> ,which</span> contains the
+
class=SpellE>codon</span> usage for <span class=SpellE>K. eikastus</span> was
-
KR2 gene, also proved quite difficult to obtain. Additionally, the <span
+
very different from <span class=SpellE>C. cresentus</span>. By entering the <span class=SpellE>codon</span> usage table for C. <span class=SpellE>cresentus</span> into an excel sheet with the KR2 DNA sequence we were able to identify the rare <span class=SpellE>codons</span> in the gene.</p>  
-
class=SpellE>codon</span> usage for <span class=SpellE>K.Eikastus</span> was
+
-
very different from <span class=SpellE>C.Cresentus</span>. In order to remedy
+
-
this, we entered in the <span class=SpellE>codon</span> usage table for C. <span
+
-
class=SpellE>Cresentus</span> on an excel sheet and created a function to find
+
-
out how many rare <span class=SpellE>codons</span> were in the original KR2
+
-
sequence along with how many rare <span class=SpellE>codons</span> were next to
+
-
each other.<o:p></o:p></span></p>
+
<p class=MsoNormal><span style='font-family:"Times New Roman";mso-fareast-font-family:
<p class=MsoNormal><span style='font-family:"Times New Roman";mso-fareast-font-family:
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<p class=MsoNormal><span style='font-family:"Times New Roman";mso-fareast-font-family:
<p class=MsoNormal><span style='font-family:"Times New Roman";mso-fareast-font-family:
-
"Times New Roman"'>Through the use of G-blocks, we specified a desired sequence
+
"Times New Roman"'>Using this information G-blocks were designed to be (<span
-
to be transformed into C. <span class=SpellE>Cresentus</span>. G-blocks (<span
+
class=SpellE>codon</span> optimized for C. <span class=SpellE>Cresentus</span> and assembled into
-
class=SpellE>codon</span> optimized for C. <span class=SpellE>Cresentus</span><span
+
the PSB1C3 standard plasmid backbone. A Gibson reaction was then performed with the G-Blocks and the  
-
class=GramE>)<span style="mso-spacerun: yes">&nbsp; </span>were</span> ordered
+
plasmid backbone to create our submission to the registry.</span><a name="_GoBack"></a></p>
-
and contained sites to fit between the prefix and suffix<span
+
-
style="mso-spacerun: yes">&nbsp; </span>with the PSB1C3 backbone . A Gibson
+
-
reaction was then performed with the G-Blocks and the plasmid backbone to
+
-
create our submission to the registry.</span><a name="_GoBack"></a></p>
+
<p class=MsoNormal><o:p>&nbsp;</o:p></p>
<p class=MsoNormal><o:p>&nbsp;</o:p></p>
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<p class=MsoNormal><o:p>&nbsp;</o:p></p>
<p class=MsoNormal><o:p>&nbsp;</o:p></p>
-
<p class=MsoNormal>We used a Gibson <span class=SpellE>reation</span> with IDT
 
-
G Blocks and IGEM pSB1C3 to clone KR2 <span class=SpellE>halorhodopsin</span>
 
-
into pSB1C3 <span class=SpellE>BioBrick</span> backbone. Results confirmed by
 
-
Sanger sequencing.</p>
 
<p class=MsoNormal><o:p>&nbsp;</o:p></p>
<p class=MsoNormal><o:p>&nbsp;</o:p></p>
-
<p class=MsoNormal>Key Reference- <span class=GramE>A</span> light-driven
+
<p class=MsoNormal>
-
sodium ion pump in marine bacteria. Inoue et al. Nature Communications DOI:
+
<h5>
-
10.1038/ncomms2689 April 9, 2013. </p>
+
Sequences:
 +
</h5>
<p class=MsoNormal><o:p>&nbsp;</o:p></p>
<p class=MsoNormal><o:p>&nbsp;</o:p></p>
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</body>
</body>
-
 
+
<h5>
 +
Key References
 +
</h5>
 +
Inoue, Keiichi et al. A light-driven sodium ion pump in marine bacteria. Nature Communications DOI:
 +
10.1038/ncomms2689 April 9, 2013. </p>
</html>
</html>

Latest revision as of 04:00, 28 September 2013

Ion Pumps

The goal of the ion pumps group was to find and obtain light driven sodium and chloride pumps to be expressed in C. cresentus. Choice of ion pumps was based on ion selectivity and structure simplicity.

 

Two pumps were used for this project. The first choice was the widely studied Halorhodopsin (HR), an inward pointing chloride conductor. For the sodium pump we chose the newly discovered KR2, an outward facing pump.

 

First, we searched the iGEM catalog for any parts containing the coding sequence for HR. We found three parts; BBaK559000, BBaK9001 and BBaK559010 and requested them for use in our project. After isolating and sequencing these plasmids, we found them to contain incomplete versions of the coding sequence of Halorhodopsin from N.pharaonis. Another problem was that the sequence of the HR in the parts contained an extra base which was not in agreement with the coding sequence the team who made the part reported. With this we searched for a new source to obtain the Halorhodopsin gene from.  

 

The next attempt was to search other organisms that contained the HR gene. We found that one of these organisms,  Halobacterium Salinarum (HS)  was in a frozen stock close by our lab on campus. Fortunately the codon usage of the wild-type HR for H. salinarum was very similar that of C. Cresentus, therefore codon optimization was not required. Specific primers were then designed to amplify the HR gene out of the HS genome. The primers contain restriction sites for ligation into our plasmids. (see tags section)

 

The second pump used in this project was a newly discovered light driven sodium pump KR2, from a lab in South Korea. The organism which contains the KR2 gene, Krokinobacter Eikastus, proved quite difficult to obtain. Additionally, the codon usage for K. eikastus was very different from C. cresentus. By entering the codon usage table for C. cresentus into an excel sheet with the KR2 DNA sequence we were able to identify the rare codons in the gene.

 

Using this information G-blocks were designed to be (codon optimized for C. Cresentus and assembled into the PSB1C3 standard plasmid backbone. A Gibson reaction was then performed with the G-Blocks and the plasmid backbone to create our submission to the registry.

 

Cloning of a Newly discovered Na pumping Halorhodopsin from marine flavobacterium Krokinobacter eikastus

 

 

Sequences:

 

DNA Sequence KR2 Na pumping halorhodopsin-

ATGACACAAGAACTAGGGAATGCCAATTTCGAAAATTTCATTGGAGCTACAG
AAGGATTTTCTGAAATTGCTTATCAATTTACATCACATATCCTTACGTTAGGGTACGCAGTGATGCTTGCAGGATT
ACTATACTTTATCCTTACCATCAAAAATGTAGATAAAAAATTCCAAATGTCGAACATATTATCAGCTGTGGTAATG
GTATCGGCATTTTTGCTATTATATGCGCAGGCACAAAACTGGACATCCAGTTTTACCTTTAATGAAGAAGTAGGAA
GATATTTTTTAGATCCGAGTGGTGATCTATTTAATAACGGATATCGCTATCTTAACTGGCTCATCGATGTACCTAT
GCTTCTCTTTCAAATTCTATTTGTAGTAAGTTTAACTACTTCAAAATTTAGCTCTGTACGTAACCAATTCTGGTTT
TCTGGGGCAATGATGATTATTACTGGGTACATTGGACAGTTTTATGAGGTAAGTAACTTGACTGCCTTTTTAGTAT
GGGGAGCTATTTCATCTGCTTTTTTCTTCCATATTTTATGGGTTATGAAGAAGGTAATTAATGAAGGAAAAGAGGG
GATTTCCCCAGCAGGACAAAAAATACTTTCTAATATCTGGATCTTATTTTTAATATCATGGACTTTATATCCAGGA
GCTTACTTAATGCCATACCTTACTGGAGTAGACGGATTTTTATATAGTGAAGATGGCGTGATGGCTAGACAACTAG
TATACACTATTGCAGATGTAAGTTCTAAAGTTATCTATGGTGTATTATTAGGTAACCTAGCAATTACATTAAGTAA
AAACAAAGAGTTGGTTGAAGCAAATAGCTAA

 

Protein Sequence KR2 Na pumping halorhodopsin-

MTQELGNANFENFIGATEGFSEIAYQFTSHILTLGYAVMLAGLLYFILTIKN
VDKKFQMSNILSAVVMVSAFLLLYAQAQNWTSSFTFNEEVGRYFLDPSGDLFNNGYRYLNWLIDVPMLLFQILFVV
SLTTSKFSSVRNQFWFSGAMMIITGYIGQFYEVSNLTAFLVWGAISSAFFFHILWVMKKVINEGKEGISPAGQKIL
SNIWILFLISWTLYPGAYLMPYLTGVDGFLYSEDGVMARQLVYTIADVSSKVIYGVLLGNLAITLSKNKELVEANS*

 

Length: 280

Molecular weight: 31.524 kDa

Isoelectric point: 4.96

 

The codon Usage table for Caulobacter crescentus-

 

Amino Acid

Codon

Number/ Genome 

#/1000

Fraction

Ala

GCG

57020

46.06

0.334895631

Ala

GCA

3967

3.2

0.023299386

Ala

GCT

10175

8.22

0.059760839

Ala

GCC

99100

80.04

0.582044144

Arg

AGG

2701

2.18

0.029839039

Arg

AGA

1059

0.86

0.011699201

Arg

CGG

19055

15.39

0.210508291

Arg

CGA

4608

3.72

0.05090644

Arg

CGT

10018

8.09

0.110672897

Arg

CGC

53078

42.87

0.586374131

Asn

AAT

5773

4.66

0.196547733

Asn

AAC

23599

19.06

0.803452267

Asp

GAT

16529

13.35

0.231087561

Asp

GAC

54998

44.42

0.768912439

Cys

TGT

1309

1.06

0.142732526

Cys

TGC

7862

6.35

0.857267474

End

TGA

1799

1.45

0.472550565

End

TAG

1247

1.01

0.327554505

End

TAA

761

0.61

0.19989493

Gln

CAG

34155

27.59

0.859829318

Gln

CAA

5568

4.5

0.140170682

Glu

GAG

47647

38.48

0.713289121

Glu

GAA

19152

15.47

0.286710879

Gly

GGG

13983

11.29

0.126510929

Gly

GGA

5358

4.33

0.048476404

Gly

GGT

11281

9.11

0.102064635

Gly

GGC

79906

64.54

0.722948031

His

CAT

6487

5.24

0.293343583

His

CAC

15627

12.62

0.706656417

Ile

ATA

790

0.64

0.014615555

Ile

ATT

3549

2.87

0.065658995

Ile

ATC

49713

40.15

0.91972545

Leu

TTG

8478

6.85

0.068407378

Leu

TTA

427

0.34

0.003445382

Leu

CTG

84529

68.27

0.68204851

Leu

CTA

1742

1.41

0.014055868

Leu

CTT

7698

6.22

0.062113706

Leu

CTC

21060

17.01

0.169929156

Lys

AAG

40138

32.42

0.924284991

Lys

AAA

3288

2.66

0.075715009

Met

ATG

26760

21.61

1

Phe

TTT

5212

4.21

0.119489213

Phe

TTC

38407

31.02

0.880510787

Pro

CCG

37265

30.1

0.552000474

Pro

CCA

2760

2.23

0.040883438

Pro

CCT

4338

3.5

0.064258099

Pro

CCC

23146

18.7

0.342857989

Ser

AGT

1687

1.36

0.026704447

Ser

AGC

20348

16.44

0.322099631

Ser

TCG

26206

21.17

0.41482912

Ser

TCA

2127

1.72

0.033669447

Ser

TCT

2144

1.73

0.03393855

Ser

TCC

10661

8.61

0.168758805

Thr

ACG

20611

16.65

0.318075896

Thr

ACA

1775

1.43

0.027392398

Thr

ACT

1768

1.43

0.027284372

Thr

ACC

40645

32.83

0.627247334

Trp

TGG

17312

13.98

1

Tyr

TAT

11882

9.6

0.459918715

Tyr

TAC

13953

11.27

0.540081285

Val

GTG

37323

30.15

0.397755611

Val

GTA

1517

1.23

0.016166848

Val

GTT

6714

5.42

0.07155189

Val

GTC

48280

39

0.514525652

 

 

Codon optimized sequence for KR2 Na pumping halorhodopsin

 

ATGACGCAAGAACTGGGGAATGCCAATTTCGAAAATTTCATCGGTGCGACCG
AAGGCTTTAGCGAAATCGCCTATCAATTTACGTCGCATATCCTCACGCTGGGGTACGCGGTGATGCTCGCCGGGCT
GCTCTACTTTATCCTGACCATCAAGAATGTCGATAAGAAGTTCCAAATGTCGAACATCCTCAGCGCGGTGGTCATG
GTGTCGGCCTTTCTGCTCCTGTATGCGCAGGCGCAAAACTGGACCTCCTCGTTTACCTTTAATGAAGAAGTCGGTC
GTTATTTTCTCGATCCGAGCGGTGATCTGTTTAATAACGGCTATCGCTATCTCAACTGGCTCATCGATGTGCCCAT
GCTGCTCTTTCAAATCCTGTTTGTCGTGTCGCTCACGACCAGCAAGTTTAGCTCGGTCCGTAACCAATTCTGGTTT
AGCGGGGCCATGATGATCATCACGGGGTACATCGGGCAGTTTTATGAGGTGTCGAACCTGACCGCCTTTCTCGTCT
GGGGTGCGATCAGCTCGGCCTTTTTCTTCCATATCCTGTGGGTGATGAAGAAGGTCATCAATGAAGGCAAGGAGGG
GATCTCCCCGGCGGGGCAAAAGATCCTCAGCAATATCTGGATCCTGTTTCTCATCTCGTGGACGCTGTATCCCGGT
GCCTACCTCATGCCGTACCTGACCGGCGTGGACGGGTTTCTCTATAGCGAAGATGGCGTGATGGCGCGCCAACTGG
TCTACACGATCGCCGATGTGTCGAGCAAGGTCATCTATGGTGTGCTCCTGGGTAACCTCGCGATCACCCTGTCGAA
GAACAAGGAGCTCGTCGAAGCCAATAGCTAA

 

Protein Sequence for codon optimized KR2 Na pumping halorhodopsin-

MTQELGNANFENFIGATEGFSEIAYQFTSHILTLGYAVMLAGLLYFILTIKN
VDKKFQMSNILSAVVMVSAFLLLYAQAQNWTSSFTFNEEVGRYFLDPSGDLFNNGYRYLNWLIDVPMLLFQILFVV
SLTTSKFSSVRNQFWFSGAMMIITGYIGQFYEVSNLTAFLVWGAISSAFFFHILWVMKKVINEGKEGISPAGQKIL
SNIWILFLISWTLYPGAYLMPYLTGVDGFLYSEDGVMARQLVYTIADVSSKVIYGVLLGNLAITLSKNKELVEANS*

 

Alignment of two protein sequences, one wild type and one codon optimized for Caulobacter (to confirm we have the same protein)

 

MTQELGNANFENFIGATEGFSEIAYQFTSHILTLGYAVMLAGLLYFILTIKN
VDKKFQMSNILSAVVMVSAFLLLYAQAQNWTSSFTFNEEVGRYFLDPSGDLFNNGYRYLNWLIDVPMLLFQILFVV
SLTTSKFSSVRNQFWFSGAMMIITGYIGQFYEVSNLTAFLVWGAISSAFFFHILWVMKKVINEGKEGISPAGQKIL
SNIWILFLISWTLYPGAYLMPYLTGVDGFLYSEDGVMARQLVYTIADVSSKVIYGVLLGNLAITLSKNKELVEANS
*

MTQELGNANFENFIGATEGFSEIAYQFTSHILTLGYAVMLAGLLYFILTIKN
VDKKFQMSNILSAVVMVSAFLLLYAQAQNWTSSFTFNEEVGRYFLDPSGDLFNNGYRYLNWLIDVPMLLFQILFVV
SLTTSKFSSVRNQFWFSGAMMIITGYIGQFYEVSNLTAFLVWGAISSAFFFHILWVMKKVINEGKEGISPAGQKIL
SNIWILFLISWTLYPGAYLMPYLTGVDGFLYSEDGVMARQLVYTIADVSSKVIYGVLLGNLAITLSKNKELVEANS
*

 

BioBrick pSB1C3 sequence with KR2 inserted- 2913 bp.

 

TACTAGTAGCGGCCGCTGCAGTCCGGCAAAAAAGGGCAAGGTGTCACCACCC
TGCCCTTTTTCTTTAAAACCGAAAAGATTACTTCGCGTTATGCAGGCTTCCTCGCTCACTGACTCGCTGCGCTCGG
TCGTTCGGCTGCGGCGAGCGGTATCAGCTCACTCAAAGGCGGTAATACGGTTATCCACAGAATCAGGGGATAACGC
AGGAAAGAACATGTGAGCAAAAGGCCAGCAAAAGGCCAGGAACCGTAAAAAGGCCGCGTTGCTGGCGTTTTTCCAC
AGGCTCCGCCCCCCTGACGAGCATCACAAAAATCGACGCTCAAGTCAGAGGTGGCGAAACCCGACAGGACTATAAA
GATACCAGGCGTTTCCCCCTGGAAGCTCCCTCGTGCGCTCTCCTGTTCCGACCCTGCCGCTTACCGGATACCTGTC
CGCCTTTCTCCCTTCGGGAAGCGTGGCGCTTTCTCATAGCTCACGCTGTAGGTATCTCAGTTCGGTGTAGGTCGTT
CGCTCCAAGCTGGGCTGTGTGCACGAACCCCCCGTTCAGCCCGACCGCTGCGCCTTATCCGGTAACTATCGTCTTG
AGTCCAACCCGGTAAGACACGACTTATCGCCACTGGCAGCAGCCACTGGTAACAGGATTAGCAGAGCGAGGTATGT
AGGCGGTGCTACAGAGTTCTTGAAGTGGTGGCCTAACTACGGCTACACTAGAAGAACAGTATTTGGTATCTGCGCT
CTGCTGAAGCCAGTTACCTTCGGAAAAAGAGTTGGTAGCTCTTGATCCGGCAAACAAACCACCGCTGGTAGCGGTG
GTTTTTTTGTTTGCAAGCAGCAGATTACGCGCAGAAAAAAAGGATCTCAAGAAGATCCTTTGATCTTTTCTACGGG
GTCTGACGCTCAGTGGAACGAAAACTCACGTTAAGGGATTTTGGTCATGAGATTATCAAAAAGGATCTTCACCTAG
ATCCTTTTAAATTAAAAATGAAGTTTTAAATCAATCTAAAGTATATATGAGTAAACTTGGTCTGACAGCTCGAGGC
TTGGATTCTCACCAATAAAAAACGCCCGGCGGCAACCGAGCGTTCTGAACAAATCCAGATGGAGTTCTGAGGTCAT
TACTGGATCTATCAACAGGAGTCCAAGCGAGCTCGATATCAAATTACGCCCCGCCCTGCCACTCATCGCAGTACTG
TTGTAATTCATTAAGCATTCTGCCGACATGGAAGCCATCACAAACGGCATGATGAACCTGAATCGCCAGCGGCATC
AGCACCTTGTCGCCTTGCGTATAATATTTGCCCATGGTGAAAACGGGGGCGAAGAAGTTGTCCATATTGGCCACGT
TTAAATCAAAACTGGTGAAACTCACCCAGGGATTGGCTGAGACGAAAAACATATTCTCAATAAACCCTTTAGGGAA
ATAGGCCAGGTTTTCACCGTAACACGCCACATCTTGCGAATATATGTGTAGAAACTGCCGGAAATCGTCGTGGTAT
TCACTCCAGAGCGATGAAAACGTTTCAGTTTGCTCATGGAAAACGGTGTAACAAGGGTGAACACTATCCCATATCA
CCAGCTCACCGTCTTTCATTGCCATACGAAATTCCGGATGAGCATTCATCAGGCGGGCAAGAATGTGAATAAAGGC
CGGATAAAACTTGTGCTTATTTTTCTTTACGGTCTTTAAAAAGGCCGTAATATCCAGCTGAACGGTCTGGTTATAG
GTACATTGAGCAACTGACTGAAATGCCTCAAAATGTTCTTTACGATGCCATTGGGATATATCAACGGTGGTATATC
CAGTGATTTTTTTCTCCATTTTAGCTTCCTTAGCTCCTGAAAATCTCGATAACTCAAAAAATACGCCCGGTAGTGA
TCTTATTTCATTATGGTGAAAGTTGGAACCTCTTACGTGCCCGATCAACTCGAGTGCCACCTGACGTCTAAGAAAC
CATTATTATCATGACATTAACCTATAAAAATAGGCGTATCACGAGGCAGAATTTCAGATAAAAAAAATCCTTAGCT
TTCGCTAAGGATGATTTCTGGAATTCGCGGCCGCTTCTAGAG/ATGACGCAAGAACTGGGGAATGCCAATTTCGAA
AATTTCATCGGTGCGACCGAAGGCTTTAGCGAAATCGCCTATCAATTTACGTCGCATATCCTCACGCTGGGGTACG
CGGTGATGCTCGCCGGGCTGCTCTACTTTATCCTGACCATCAAGAATGTCGATAAGAAGTTCCAAATGTCGAACAT
CCTCAGCGCGGTGGTCATGGTGTCGGCCTTTCTGCTCCTGTATGCGCAGGCGCAAAACTGGACCTCCTCGTTTACC
TTTAATGAAGAAGTCGGTCGTTATTTTCTCGATCCGAGCGGTGATCTGTTTAATAACGGCTATCGCTATCTCAACT
GGCTCATCGATGTGCCCATGCTGCTCTTTCAAATCCTGTTTGTCGTGTCGCTCACGACCAGCAAGTTTAGCTCGGT
CCGTAACCAATTCTGGTTTAGCGGGGCCATGATGATCATCACGGGGTACATCGGGCAGTTTTATGAGGTGTCGAAC
CTGACCGCCTTTCTCGTCTGGGGTGCGATCAGCTCGGCCTTTTTCTTCCATATCCTGTGGGTGATGAAGAAGGTCA
TCAATGAAGGCAAGGAGGGGATCTCCCCGGCGGGGCAAAAGATCCTCAGCAATATCTGGATCCTGTTTCTCATCTC
GTGGACGCTGTATCCCGGTGCCTACCTCATGCCGTACCTGACCGGCGTGGACGGGTTTCTCTATAGCGAAGA
TGGCGTGATGGCGCGCCAACTGGTCTACACGATCGCCGATGTGTCGAGCAAGGTCATCTATGGTGTGCTCCTGGGT
AACCTCGCGATCACCCTGTCGAAGAACAAGGAGCTCGTCGAAGCCAATAGCTAA/

Key References
Inoue, Keiichi et al. A light-driven sodium ion pump in marine bacteria. Nature Communications DOI: 10.1038/ncomms2689 April 9, 2013.

Retrieved from "http://2013.igem.org/Team:UC-Santa_Cruz/Project/Pumps"