Hybrid Protein

Brian Naughton | Sat 16 August 2014 | biotech | DNA vaccine

Starting with a protein sequence, there are several ways to produce a DNA sequence with codons that are optimized for expression. I created a DNA sequence for my hybrid flagellin using Life Tech GeneOptimizer. I selected Mus musculus as the target organism and I also selected the option to add a Kozak sequence. Here's what gets spit out.

GCCACCATGGCCCAAGTGATCAACACCAACAGCCTGAGCCTGATCACCCAGAACAACATCAACAAGAACCAGAGCGCCCTGAGCAGCAGCATCGAGAGAC
TGAGCAGCGGCCTGAGAATCAACAGCGCCAAGGACGATGCCGCTGGCCAGGCTATCGCCAACAGATTCACCAGCAACATCAAGGGCCTGACCCAGGCCGC
CAGAAACGCCAACGATGGCATCAGCGTGGCCCAGACAACAGAGGGCGCACTGAGCGAGATCAACAACAACCTGCAGAGAGTGCGCGAGCTGACCGTGCAG
GCCACCACAGGCACAAACAGCGAGAGCGACCTGTCCAGCATCCAGGACGAGATCAAGAGCAGACTGGATGAGATCGACAGAGTGTCCGGCCAGACCCAGT
TCAACGGCGTGAACGTGCTGGCCAAGAACGGCAGCATGAAGATCCAAGTGGGCGCCAACGACAACCAGACCATCACCATCGACCTGAAGCAGATCGACGC
CAAGACCCTGGGCCTGGACGGCTTCTCTGTGAAGGCCCTGATCACAGCCAGCGGCGACATCAGCCTGACCTTCAAACAGGTGGACGGCGTGAACGACGTG
ACCCTGGAAAGCGTGAAGGTGTCCAGCTCTGCCGGCACAGGCATCGGCGTGCTGGCTGAAGTGATTAACAAGAACAGCAACAGGACCGGCGTGAAGGCCT
ACGCCAGCGTGATCACCACAAGCGACGTGGCCGTGCAGAGCGGCAGCCTGTCTAACCTGACCCTGAACGGCATCCACCTGGGCAATATCGCCGATATCAA
GAAGAACGACAGCGACGGCAGACTGGTGGCCGCCATCAACGCCGTGACAAGCGAGACAGGCGTGGAAGCCTACACCGACCAGAAGGGCAGACTGAACCTG
AGATCCATCGACGGCAGGGGCATCGAAATCAAGACCGACAGCGTGTCCAACGGCCCCTCCGCCCTGACAATGGTCAACGGCGGACAGGACCTGACCAAGG
GCAGCACCAACTACGGCAGGCTGTCCCTGACAAGACTGGACGCCAAGTCCATCAACGTGGTGTCCGCCAGCGACTCCCAGCACCTGGGCTTTACAGCTAT
CGGCTTCGGCGAGAGCCAGGTGGCCGAGACTACCGTGAATCTGAGGGACGTGACCGGCAACTTCAACGCTAACGTGAAGTCCGCCTCCGGCGCTAACTAC
AACGCTGTGATCGCCTCTGGCAACCAGAGCCTGGGAAGCGGCGTGACCACACTGAGAGCTGTGGCCAACGGCAAGACCACCGACCCCCTGAAGGCTCTGG
ACGACGCTATCGCCTCCGTGGACAAGTTCAGATCCAGCCTGGGCGCTGTGCAGAACAGACTGGACAGCGCCGTGACCAACCTGAACAACACCACCACCAA
CCTGTCCGAGGCCCAGAGCAGAATCCAGGATGCCGACTACGCCACCGAGGTGTCCAACATGAGCAAGGCCCAGATCATCCAGCAGGCCGGCAACTCTGTG
CTGGCTAAGGCCAACCAGGTGCCCCAGCAGGTGCTGAGTCTGCTGCAGGGA

When I align my sequence with the original BioBrick created for the Slovenian iGem project, the two DNA sequences are only about 75% concordant (the protein sequences were identical). I expected some differences but not this many. My best guess is that the Slovenian BioBrick is not intended for use in mouse, but I really don't know what is going on. It would be interesting to test the difference in expression between the two sequences in a mouse.

CLUSTAL O(121) multiple sequence alignment

LifeTech              GCCACCATGGCCCAAGTGATCAACACCAACAGCCTGAGCCTGATCACCCAGAACAACATC
Slovenia              ------atggcacaagtcattaataccaacagcctctcgctgatcactcaaaataatatc
                            ***** ***** ** ** ***********    ******** ** ** ** ***

LifeTech              AACAAGAACCAGAGCGCCCTGAGCAGCAGCATCGAGAGACTGAGCAGCGGCCTGAGAATC
Slovenia              aacaagaaccagtctgcgctgtcgagttctatcgagcgtctgtcttctggcttgcgtatt
                      ************   ** ***   **    ****** * ***      *** ** * **

LifeTech              AACAGCGCCAAGGACGATGCCGCTGGCCAGGCTATCGCCAACAGATTCACCAGCAACATC
Slovenia              aacagcgcgaaggatgacgcagcgggtcaggcgattgctaaccgtttcacctctaacatt
                      ******** ***** ** ** ** ** ***** ** ** *** * ******   *****

LifeTech              AAGGGCCTGACCCAGGCCGCCAGAAACGCCAACGATGGCATCAGCGTGGCCCAGACAACA
Slovenia              aaaggcctgactcaggcggcccgtaacgccaacgacggtatctccgttgcgcagaccacc
                      ** ******** ***** *** * *********** ** ***  *** ** ***** **

LifeTech              GAGGGCGCACTGAGCGAGATCAACAACAACCTGCAGAGAGTGCGCGAGCTGACCGTGCAG
Slovenia              gaaggcgcgctgtccgaaatcaacaacaacttacagcgtgtgcgtgaactgacggtacag
                      ** ***** ***  *** ************ * *** * ***** ** ***** ** ***

LifeTech              GCCACCACAGGCACAAACAGCGAGAGCGACCTGTCCAGCATCCAGGACGAGATCAAGAGC
Slovenia              gccactaccggtactaactctgagtctgatctgtcttctatccaggacgaaattaaatcc
                      ***** ** ** ** ***   ***   ** *****    *********** ** **   *

LifeTech              AGACTGGATGAGATCGACAGAGTGTCCGGCCAGACCCAGTTCAACGGCGTGAACGTGCTG
Slovenia              cgtctggatgaaattgaccgcgtatctggtcagacccagttcaacggcgtgaacgtgctg
                      * ******** ** *** * ** ** ** ******************************

LifeTech              GCCAAGAACGGCAGCATGAAGATCCAAGTGGGCGCCAACGACAACCAGACCATCACCATC
Slovenia              gcaaaaaatggctccatgaaaatccaggttggcgcaaatgataaccagactatcactatc
                      ** ** ** ***  ****** ***** ** ***** ** ** ******** ***** ***

LifeTech              GACCTGAAGCAGATCGACGCCAAGACCCTGGGCCTGGACGGCTTCTCTGTGAAGGCCCTG
Slovenia              gatctgaagcagattgatgctaaaactcttggccttgatggttttagcgttaaagcgtta
                      ** *********** ** ** ** ** ** ***** ** ** **    ** ** **  *

LifeTech              ATCACAGCCAGCGGCGACATCAGCCTGACCTTCAAACAGGTGGACGGCGTGAACGACGTG
Slovenia              atcacggcttctggggatattagcttgacttttaaacaagtggatggcgtgaatgatgta
                      ***** **    ** ** ** *** **** ** ***** ***** ******** ** **

LifeTech              ACCCTGGAAAGCGTGAAGGTGTCCAGCTCTGCCGGCACAGGCATCGGCGTGCTGGCTGAA
Slovenia              actttagagagcgtaaaagtttctagttcagcaggcacggggatcggtgtgttagcggaa
                      **  * ** ***** ** ** ** ** ** ** ***** ** ***** *** * ** ***

LifeTech              GTGATTAACAAGAACAGCAACAGGACCGGCGTGAAGGCCTACGCCAGCGTGATCACCACA
Slovenia              gtgattaacaaaaattctaaccgaacaggggttaaagcttatgcgagcgttatcaccacg
                      *********** **    *** * ** ** ** ** ** ** ** ***** ********

LifeTech              AGCGACGTGGCCGTGCAGAGCGGCAGCCTGTCTAACCTGACCCTGAACGGCATCCACCTG
Slovenia              agcgatgtggcggtccaatcaggaagtttgagtaatttaactttaaatgggatccatttg
                      ***** ***** ** **    ** **  **  ***  * **  * ** ** *****  **

LifeTech              GGCAATATCGCCGATATCAAGAAGAACGACAGCGACGGCAGACTGGTGGCCGCCATCAAC
Slovenia              ggtaatatcgcagatattaagaaaaatgactcagacggaaggttagtcgcagcgatcaat
                      ** ******** ***** ***** ** ***   ***** **  * ** ** ** *****

LifeTech              GCCGTGACAAGCGAGACAGGCGTGGAAGCCTACACCGACCAGAAGGGCAGACTGAACCTG
Slovenia              gcggttacttcagaaaccggcgtggaagcttatacggatcaaaaagggcgcttgaatttg
                      ** ** **    ** ** *********** ** ** ** ** ** **  *  ****  **

LifeTech              AGATCCATCGACGGCAGGGGCATCGAAATCAAGACCGACAGCGTGTCCAACGGCCCCTCC
Slovenia              cgcagtatagatggtcgtgggattgaaatcaaaaccgatagcgtcagtaatgggcctagt
                      *    ** ** **  * ** ** ******** ***** *****    ** ** **

LifeTech              GCCCTGACAATGGTCAACGGCGGACAGGACCTGACCAAGGGCAGCACCAACTACGGCAGG
Slovenia              gctttaacgatggtcaatggcggtcaggatttaacaaaaggttctactaactatgggagg
                      **  * ** ******** ***** *****  * ** ** **    ** ***** ** ***

LifeTech              CTGTCCCTGACAAGACTGGACGCCAAGTCCATCAACGTGGTGTCCGCCAGCGACTCCCAG
Slovenia              ctttctctcacacgcttagacgctaaaagcatcaatgtcgtttcggcttctgattcgcaa
                      ** ** ** *** *  * ***** **   ****** ** ** ** **    ** ** **

LifeTech              CACCTGGGCTTTACAGCTATCGGCTTCGGCGAGAGCCAGGTGGCCGAGACTACCGTGAAT
Slovenia              catttaggtttcacagcgattggttttggggaatctcaagtggcagaaaccacggtgaat
                      **  * ** ** ***** ** ** ** ** **    ** ***** ** ** ** ******

LifeTech              CTGAGGGACGTGACCGGCAACTTCAACGCTAACGTGAAGTCCGCCTCCGGCGCTAACTAC
Slovenia              ttgcgcgatgttactgggaattttaacgctaatgtcaaatcagccagtggcgcgaactat
                      ** * ** ** ** ** ** ** ******** ** ** ** ***   ***** *****

LifeTech              AACGCTGTGATCGCCTCTGGCAACCAGAGCCTGGGAAGCGGCGTGACCACACTGAGAGCT
Slovenia              aacgccgtgatcgctagcggtaaccaaagcttgggatctggggttacaaccttaagagct
                      ***** ********    ** ***** *** *****   ** ** ** **  * ******

LifeTech              GTGGCCAACGGCAAGACCACCGACCCCCTGAAGGCTCTGGACGACGCTATCGCCTCCGTG
Slovenia              gttgcaaatggtaaaaccacggatccgctgaaagcgctggacgatgctatcgcatctgta
                      ** ** ** ** ** ***** ** ** ***** ** ******** ******** ** **

LifeTech              GACAAGTTCAGATCCAGCCTGGGCGCTGTGCAGAACAGACTGGACAGCGCCGTGACCAAC
Slovenia              gacaaattccgttcttccctcggtgcggtgcaaaaccgtctggattccgcggttaccaac
                      ***** *** * **   *** ** ** ***** *** * *****   *** ** ******

LifeTech              CTGAACAACACCACCACCAACCTGTCCGAGGCCCAGAGCAGAATCCAGGATGCCGACTAC
Slovenia              ctgaacaacaccactaccaacctgtctgaagcgcagtcccgtattcaggacgccgactat
                      ************** *********** ** ** ***  * * ** ***** ********

LifeTech              GCCACCGAGGTGTCCAACATGAGCAAGGCCCAGATCATCCAGCAGGCCGGCAACTCTGTG
Slovenia              gcgaccgaagtgtccaatatgtcgaaagcgcagatcatccagcaggccggtaactccgtg
                      ** ***** ******** ***   ** ** ******************** ***** ***

LifeTech              CTGGCTAAGGCCAACCAGGTGCCCCAGCAGGTGCTGAGTCTGCTGCAGGGA---------
Slovenia              ttggcaaaagctaaccaggtaccgcagcaggttctgtctctgttacagggttactagagc
                      **** ** ** ******** ** ******** ***  **** * *****

LifeTech              -----------------------------
Slovenia              gaggagaccaccaccaccaccaccactag

There are fewer restriction sites in the sequence generated by GeneOptimizer. I don't think that's proof of anything, but seems generally positive. (The images below are from the NEBcutter online tool.)

Comparing Codon Optimizers

I compared three methods of codon optimization: JCAT, IDT and GeneOptimizer. In all three cases I select for Mus musculus as the target organism. There is good concordance between all three, with LifeTech and JCAT at over 90% condordance. Note the addiitonal leader sequence in the LifeTech version — that's the added Kozak sequence.

CLUSTAL O(1.2.1) multiple sequence alignment


IDT           ------ATGGCACAGGTCATTAATACTAATAGTCTGAGTCTGATCACTCAGAATAACATC
LifeTech      GCCACCATGGCCCAAGTGATCAACACCAACAGCCTGAGCCTGATCACCCAGAACAACATC
JCAT          ------ATGGCCCAGGTGATCAACACCAACAGCCTGAGCCTGATCACCCAGAACAACATC
                    *****.**.** ** ** ** ** ** ***** ******** ***** ******

IDT           AACAAAAACCAATCAGCCCTGTCAAGTTCCATAGAGAGGCTGAGTAGCGGACTGAGGATA
LifeTech      AACAAGAACCAGAGCGCCCTGAGCAGCAGCATCGAGAGACTGAGCAGCGGCCTGAGAATC
JCAT          AACAAGAACCAGAGCGCCCTGAGCAGCAGCATCGAGAGGCTGAGCAGCGGCCTGAGGATC
              *****.*****.: .******: .** : ***.*****.***** *****.*****.**.

IDT           AACTCAGCAAAAGATGATGCTGCTGGTCAGGCTATAGCCAACCGGTTCACGTCTAACATA
LifeTech      AACAGCGCCAAGGACGATGCCGCTGGCCAGGCTATCGCCAACAGATTCACCAGCAACATC
JCAT          AACAGCGCCAAGGACGACGCCGCCGGCCAGGCCATCGCCAACAGGTTCACCAGCAACATC
              ***: .**.**.** ** ** ** ** ***** **.******.*.***** :  *****.

IDT           AAAGGATTGACCCAGGCTGCCCGGAACGCTAATGATGGAATCAGCGTGGCCCAGACTACA
LifeTech      AAGGGCCTGACCCAGGCCGCCAGAAACGCCAACGATGGCATCAGCGTGGCCCAGACAACA
JCAT          AAGGGCCTGACCCAGGCCGCCAGGAACGCCAACGACGGCATCAGCGTGGCCCAGACCACC
              **.**. ********** ***.*.***** ** ** **.***************** **.

IDT           GAAGGCGCTTTGAGTGAAATCAACAATAACCTCCAAAGAGTGAGAGAATTGACTGTGCAG
LifeTech      GAGGGCGCACTGAGCGAGATCAACAACAACCTGCAGAGAGTGCGCGAGCTGACCGTGCAG
JCAT          GAGGGCGCCCTGAGCGAGATCAACAACAACCTGCAGAGGGTGAGGGAGCTGACCGTGCAG
              **.*****  **** **.******** ***** **.**.***.* **. **** ******

IDT           GCTACTACTGGAACAAATAGCGAGTCCGATCTGTCTAGCATCCAGGATGAAATTAAATCC
LifeTech      GCCACCACAGGCACAAACAGCGAGAGCGACCTGTCCAGCATCCAGGACGAGATCAAGAGC
JCAT          GCCACCACCGGCACCAACAGCGAGAGCGACCTGAGCAGCATCCAGGACGAGATCAAGAGC
              ** ** ** **.**.** ******: *** ***:  *********** **.** **.: *

IDT           AGACTGGATGAAATCGATCGCGTGAGCGGACAAACTCAGTTTAACGGAGTGAATGTTCTG
LifeTech      AGACTGGATGAGATCGACAGAGTGTCCGGCCAGACCCAGTTCAACGGCGTGAACGTGCTG
JCAT          AGGCTGGACGAGATCGACAGGGTGAGCGGCCAGACCCAGTTCAACGGCGTGAACGTGCTG
              **.***** **.***** .* ***: ***.**.** ***** *****.***** ** ***

IDT           GCTAAAAATGGCAGTATGAAGATCCAGGTCGGAGCCAACGACAACCAGACAATTACAATC
LifeTech      GCCAAGAACGGCAGCATGAAGATCCAAGTGGGCGCCAACGACAACCAGACCATCACCATC
JCAT          GCCAAGAACGGCAGCATGAAGATCCAGGTGGGCGCCAACGACAACCAGACCATCACCATC
              ** **.** ***** ***********.** **.*****************.** **.***

IDT           GACCTCAAGCAAATCGACGCAAAGACGCTGGGCCTGGATGGATTTTCTGTGAAGGCATTG
LifeTech      GACCTGAAGCAGATCGACGCCAAGACCCTGGGCCTGGACGGCTTCTCTGTGAAGGCCCTG
JCAT          GACCTGAAGCAGATCGACGCCAAGACCCTGGGCCTGGACGGCTTCAGCGTGAAGGCCCTG
              ***** *****.********.***** *********** **.** :  ********. **

IDT           ATAACCGCCTCCGGCGATATTTCTTTGACTTTCAAACAGGTCGACGGGGTCAACGACGTC
LifeTech      ATCACAGCCAGCGGCGACATCAGCCTGACCTTCAAACAGGTGGACGGCGTGAACGACGTG
JCAT          ATCACCGCCAGCGGCGACATCAGCCTGACCTTCAAGCAGGTGGACGGCGTGAACGACGTG
              **.**.***: ****** ** :   **** *****.***** ***** ** ********

IDT           ACCTTGGAAAGCGTAAAAGTGTCATCATCAGCAGGCACAGGGATTGGCGTGCTGGCTGAG
LifeTech      ACCCTGGAAAGCGTGAAGGTGTCCAGCTCTGCCGGCACAGGCATCGGCGTGCTGGCTGAA
JCAT          ACCCTGGAGAGCGTGAAGGTGAGCAGCAGCGCCGGCACCGGCATCGGCGTGCTGGCCGAG
              *** ****.*****.**.***: .: .:  **.*****.** ** *********** **.

IDT           GTGATCAACAAGAATTCCAATAGGACCGGAGTGAAGGCCTATGCATCAGTGATAACTACC
LifeTech      GTGATTAACAAGAACAGCAACAGGACCGGCGTGAAGGCCTACGCCAGCGTGATCACCACA
JCAT          GTGATCAACAAGAACAGCAACAGGACCGGCGTGAAGGCCTACGCCAGCGTGATCACCACC
              ***** ******** : *** ********.*********** **.: .*****.** **.

IDT           TCCGATGTGGCTGTCCAGTCCGGGTCCCTGAGCAATCTGACCCTGAACGGCATTCACCTG
LifeTech      AGCGACGTGGCCGTGCAGAGCGGCAGCCTGTCTAACCTGACCCTGAACGGCATCCACCTG
JCAT          AGCGACGTGGCCGTGCAGAGCGGCAGCCTGAGCAACCTGACCCTGAACGGCATCCACCTG
              : *** ***** ** ***: *** : ****:  ** ***************** ******

IDT           GGCAACATCGCAGATATTAAAAAGAACGACTCTGACGGACGCCTTGTGGCAGCAATCAAT
LifeTech      GGCAATATCGCCGATATCAAGAAGAACGACAGCGACGGCAGACTGGTGGCCGCCATCAAC
JCAT          GGCAACATCGCCGACATCAAGAAGAACGACAGCGACGGCAGGCTGGTGGCCGCCATCAAC
              ***** *****.** ** **.*********:  *****..* ** *****.**.*****

IDT           GCTGTCACATCCGAGACTGGAGTGGAAGCCTACACTGATCAGAAAGGCCGACTTAACTTG
LifeTech      GCCGTGACAAGCGAGACAGGCGTGGAAGCCTACACCGACCAGAAGGGCAGACTGAACCTG
JCAT          GCCGTGACCAGCGAGACCGGCGTGGAGGCCTACACCGACCAGAAGGGCAGGCTGAACCTG
              ** ** **.: ****** **.*****.******** ** *****.***.*.** *** **

IDT           AGGTCTATCGACGGAAGAGGTATCGAAATCAAGACCGATAGTGTGTCCAATGGCCCCTCC
LifeTech      AGATCCATCGACGGCAGGGGCATCGAAATCAAGACCGACAGCGTGTCCAACGGCCCCTCC
JCAT          AGGAGCATCGACGGCAGGGGCATCGAGATCAAGACCGACAGCGTGAGCAACGGCCCCAGC
              **.:  ********.**.** *****.*********** ** ***: *** ******: *

IDT           GCACTTACTATGGTGAACGGGGGACAGGATCTCACCAAAGGGAGCACCAACTATGGCAGA
LifeTech      GCCCTGACAATGGTCAACGGCGGACAGGACCTGACCAAGGGCAGCACCAACTACGGCAGG
JCAT          GCCCTGACCATGGTGAACGGCGGCCAGGACCTGACCAAGGGCAGCACCAACTACGGCAGG
              **.** ** ***** ***** **.***** ** *****.** *********** *****.

IDT           CTGTCATTGACTAGACTGGATGCTAAGAGCATCAATGTTGTGTCTGCTTCTGACTCCCAA
LifeTech      CTGTCCCTGACAAGACTGGACGCCAAGTCCATCAACGTGGTGTCCGCCAGCGACTCCCAG
JCAT          CTGAGCCTGACCAGGCTGGACGCCAAGAGCATCAACGTGGTGAGCGCCAGCGACAGCCAG
              ***: . **** **.***** ** ***: ****** ** ***:  ** :  ***: ***.

IDT           CATCTGGGCTTCACCGCCATTGGTTTCGGCGAGTCCCAGGTCGCCGAAACTACTGTAAAT
LifeTech      CACCTGGGCTTTACAGCTATCGGCTTCGGCGAGAGCCAGGTGGCCGAGACTACCGTGAAT
JCAT          CACCTGGGCTTCACCGCCATCGGCTTCGGCGAGAGCCAGGTGGCCGAGACCACCGTGAAC
              ** ******** **.** ** ** *********: ****** *****.** ** **.**

IDT           TTGCGAGATGTGACCGGAAATTTTAATGCCAACGTCAAATCAGCCAGTGGCGCAAACTAC
LifeTech      CTGAGGGACGTGACCGGCAACTTCAACGCTAACGTGAAGTCCGCCTCCGGCGCTAACTAC
JCAT          CTGAGGGACGTGACCGGCAACTTCAACGCCAACGTGAAGAGCGCCAGCGGCGCCAACTAC
              **.*.** ********.** ** ** ** ***** **.: .***:  ***** ******

IDT           AATGCTGTGATCGCTAGTGGAAACCAATCTCTTGGCAGTGGGGTAACTACTCTGAGGGCT
LifeTech      AACGCTGTGATCGCCTCTGGCAACCAGAGCCTGGGAAGCGGCGTGACCACACTGAGAGCT
JCAT          AACGCCGTGATCGCCAGCGGCAACCAGAGCCTGGGCAGCGGCGTGACCACCCTGAGGGCC
              ** ** ******** :  **.*****.:  ** **.** ** **.** ** *****.**

IDT           GTGGCCAACGGCAAGACCACAGATCCCCTGAAGGCCCTCGATGACGCAATAGCCTCTGTG
LifeTech      GTGGCCAACGGCAAGACCACCGACCCCCTGAAGGCTCTGGACGACGCTATCGCCTCCGTG
JCAT          GTGGCCAACGGCAAGACCACCGACCCCCTGAAGGCCCTGGACGACGCCATCGCCAGCGTG
              ********************.** *********** ** ** ***** **.***:  ***

IDT           GATAAATTTAGAAGTAGCCTTGGAGCCGTGCAGAATCGACTCGATAGCGCTGTTACAAAT
LifeTech      GACAAGTTCAGATCCAGCCTGGGCGCTGTGCAGAACAGACTGGACAGCGCCGTGACCAAC
JCAT          GACAAGTTCAGGAGCAGCCTGGGCGCCGTGCAGAACAGGCTGGACAGCGCCGTGACCAAC
              ** **.** **.:  ***** **.** ******** .*.** ** ***** ** **.**

IDT           CTGAATAACACCACCACAAATCTGTCTGAAGCACAGTCACGCATTCAGGATGCTGATTAT
LifeTech      CTGAACAACACCACCACCAACCTGTCCGAGGCCCAGAGCAGAATCCAGGATGCCGACTAC
JCAT          CTGAACAACACCACCACCAACCTGAGCGAGGCCCAGAGCAGGATCCAGGACGCCGACTAC
              ***** ***********.** ***:  **.**.***: ..* ** ***** ** ** **

IDT           GCAACAGAAGTTAGCAATATGTCCAAGGCCCAAATCATCCAACAGGCTGGGAACAGCGTC
LifeTech      GCCACCGAGGTGTCCAACATGAGCAAGGCCCAGATCATCCAGCAGGCCGGCAACTCTGTG
JCAT          GCCACCGAGGTGAGCAACATGAGCAAGGCCCAGATCATCCAGCAGGCCGGCAACAGCGTG
              **.**.**.** : *** ***: *********.********.***** ** ***:  **

IDT           CTTGCCAAAGCTAATCAGGTACCACAGCAGGTATTGTCCCTGCTTCAGGGG
LifeTech      CTGGCTAAGGCCAACCAGGTGCCCCAGCAGGTGCTGAGTCTGCTGCAGGGA
JCAT          CTGGCCAAGGCCAACCAGGTGCCCCAGCAGGTGCTGAGCCTGCTGCAGGGC
              ** ** **.** ** *****.**.********. **:  ***** *****

Hybrid Protein

Comparing Codon Optimizers

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