The thirty three sequences below were designed with the R2oDNA designer tool to be biologically inert.

>s1 ❶
AATCCAATCAGCGTAAGGTGTAGACTTTCT
>s2 ❷
ATCGTATCTGCTGCGTAAATAGTAGTCAAC
>s3 ❸
TAAAATCTCGTAAAGGAACTGTCTGCTCTG
>s4 ❹
AACTGTAAAATCAGGTATCTCGTAGTCCGT
>s5 ❺
GAAAAGCGTTTACCTCGGAACTCTATTGTA
>6
TTTACCTGTGGATTGATAGGAATACACCCA
>7
TTGAGATAGCAATAAATCCTGGTTCCGACT
>8
TAAACACAGTAACGGATTGCTTTCGTGCTA
>9
CTCGTGAATCTGGGAAATCCTTATCGTAAA
>10
AGTTATTACGAGTATTCAACCTGACGGCTA
>11
TGGTAACTGCGAACCTACTGTAAGATTCTA
>12
GTAGGTATTGAACCCGAAAATCGTATCTTC
>13
ATAATCTATCTGTAGGACCTGGTAGTCACA
>14
AAAGTATTCAGCGACTATTGGCTTCTACGA
>15
GTAGTTTCTTACAATAACGCCGATACAGGT
>16
AGGTAAAATCGGTCCCTTATCTACTGTGAA
>17
AAGATTGGTATCCCTCACGACTGTAGTAAT
>18
GTCGTAAATACAATCTCGGAATAGGCTCTT
>19
TGTAAAGTGGTAATCTTCAATCCAGAGCCT
>20
AATCTCTTCCTGGTGACACTGAATAAGGTA
>21
TGATAGCGAGTTGCCTCCAATAAAATCGTT
>22
CTTTATTTCGTAGACGCTATCAGAGACAGA
>23
ATAAGTGTCTCCCGTTGATTCTGAAAAGCA
>24
GTGAGTAGACTTATTGTTTACCCAACCGAA
>25
GAGAGTATTTACAACGCAGTCTTCGTCAAT
>26
AGGTGCTCTTATTATCGGACAGTCAATCAA
>27
GAAACTTATCGTTCAATAGGGTTCCCGTAA
>28
TCAGAAGTCTCCAATAAGGTTGTGATACTC
>29
AGAGGTTTTATTACAACAGGTCCACTTCAG
>30
ATCTGCGGTATCTCAGTAATAAGAATCGCT
>31
TAGGGCACTCTACAATAATCTTCTGGTAAG
>32
ATTATTGGGTGAACCACACGAGTAACCTTT
>33
TGGGTTCAACAATAAACTTTCGCAGGTATC

The text below was generated from the software tool:

ProjectName	
JobID r2o_6389398447409610491
Email bjorn_johansson@bio.uminho.pt
Seqnum 6
Format NNNNNNNNNNNNNNNNNNNNNNNNNNNNNN
ReverseMode false
Evalue 1
Genomes saccharomyces_cerevisiae_genome eschericichia_coli_k12_dh10b eschericichia_coli_k12_mg1655 eschericichia_coli_k12_w3110 bacillus_subtilis_168 igem_all_parts_082013 
gc 0.4
MinInterEnergy -9.0
MinIntraEnergy -4.0
FoldTemp 50.0
MaxSWScore 90.0
MaxSubMatch 10
ForbSeq TTGACA	E.coli sig70 -35 site
ForbSeq TATAAT	E.coli sig70 -10 site
ForbSeq TTGNNNNNNNNNNNNNNNNNNNNTATNNT	E.coli sig70 promoter weak consensus
ForbSeq TGGCACGNNNNTTGC	E.coli sig54 promoter consensus
ForbSeq TCNCCCTTGAANNNNNNNNNNNNNNNNNCCCCATTTA	E.coli sig32 promoter consensus
ForbSeq GAACTNNNNNNNNNNNNNNNNGTCNNA	E.coli sig24 promoter consensus
ForbSeq AAAGA	RBS
ForbSeq AGGAGG	Shine-Dalgarno sequence or 2xArg bad codon
ForbSeq ATG	translation initiation sites
ForbSeq TTATNCACA	DnaA binding sites
ForbSeq TGTGANNNNNNTCACANT	CAP binding sites
ForbSeq CTAG	IS5 insertion site
ForbSeq CTAA	IS5 insertion site
ForbSeq CAAA	IS5 insertion site
ForbSeq CAAG	IS5 insertion site
ForbSeq NGCTNAGCN	IS10 insertion site
ForbSeq GGGNNNNNCCC	IS231 insertion site
ForbSeq G{3,}[ATGC]{1,7}G{3,}[ATGC]{1,7}G{3,}[ATGC]{1,7}G{3,}	G-quadruplex
ForbSeq GGGG	G-quadruplex
ForbSeq GAATTC	EcoRI
ForbSeq ACTAGT	SpeI
ForbSeq TCTAGA	XbaI
ForbSeq CTGCAG	PstI
ForbSeq GGTCTC	BsaI
ForbSeq AAAAA	run
ForbSeq TTTTT	run
ForbSeq GGGGG	run
ForbSeq CCCCC	run
ForbSeq ATATAT	run
ForbSeq ACACAC	run
ForbSeq AGAGAG	run
ForbSeq TATATA	run
ForbSeq TCTCTC	run
ForbSeq TGTGTG	run
ForbSeq CACACA	run
ForbSeq CTCTCT	run
ForbSeq CGCGCG	run
ForbSeq GAGAGA	run
ForbSeq GTGTGT	run
ForbSeq GCGCGC	run
ClusterID 4206134 


sequencesStep1File.fa: 60	#after initial sequence generation
sequencesStep1File_pf_self.fa: 60	#after folding and self-annealing energy filter
sequencesStep2File.fa: 43	#after BLAST elimination
sequencesFinalFile.fa: 33	#final orthogonal sequence count after network elimination