Metagenome simulation from a concatenated FASTA file

Question

I am trying to simulate metagenome sequencing. So I will start with a file with a lot of concatenated genomes. From there, I would like to randomly extract 10,000 sequences of length 200bps.

I got help subtracting a random 200 sequence from a fasta file like this:

>JQ086372.1 Enterobacteria phage HK446, complete genome 
ATGGCAGATAAACGAATCCGTTCCGACAGTTCGGCGGCAGCGGTTCAGGCCATGAAAAATGCAGCTGTGGCTTCTCCCGCTTCAGTCTGCCAATATGGATGTGAAACTCGTCGGAAAAAAAGTGGTTTATCGCTATCAACGCAACAGCGAATACGCCGACTTTTCGCAGAAAGAGATTTTTCACCTTAAAGGCTTCGGATTCACCGGGCTTGTAGGCCTGTCAATTTTTTCCACTTATTTAAATGTTTGTAAATTAAGTAAATAACGTTTTTAATCTAAAAATTCCACAATTTCCACAATAACTACTACTACTAAATTTTTTTAATTATTTTATTTATTTATATTAGAAAAGAGAACAATATAAGTTATTGAATTTAATTTACGAAAAAAGTATATTGTGTATGTGTTAAAAACTTCCAAATTCCGATGGAAATTATTTGTAATCAAAATGAGTTAAACAATGCTATACAACTAGTAAGTAAAGCAGTTGCTTCACGACCAACTCATCCAATTCTTGCAAATATACTTTTAACAGCTGACCAAGGAACTAATAAAATTAGTTTGACAGGATTTGATCTTAATTTAGGAATTCAAACTTCTTTTGATGGAACTGTTAAAAATAGCGGTGCTATTACGATACCTTCAAAACTTTTATCTGAAATAGTAAACAAATTAC...(this still goes for a lot more A or C or G or T)

using this:

tail -n +2 file.fasta | tr -d '\n' > newfile
n=$(stat -c "%s" newfile)
r=$(shuf -i1-"$((n-200+1))" -n1)
newfile tail -c+"$r" | head -c200

And as an output I got: GTTAAGGGCGCCGTTCTGCTTGTCTTTGCTGACATGTTTGAACACCGGACGGCACAAAGCGAAGTGCAGCTTTATGAGAATGCAGCCGCCGAACGCATGATGTTCATTCATCGCAACTGGCGCGGTAAATCTGAACCTGAGGAGGGCTCCTGATGGAACCTGGACGATTCAGGCACAGGGTAAAAATTCTCACCTTCACG

But now I want to extract instead of 1 fragment of 200 characters; 10,0000 random fragments of 200 characters using the strategy above. For this, I am looking to exclude all those lines that begin with ">" (p.e. >JQ086372.1 Enterobacteria phage HK446, complete genome).The input file looks like this:

>JQ086372.1 Enterobacteria phage HK446, complete genome 
ATGGCAGATAAACGAATCCGTTCCGACAGTTCGGCGGCAGCGGTTCAGGCCATGAAAAATGCAGCTGTGGCTTCTCCCGCTTCAGTCTGCCAATATGGATGTGAAACTCGTCGGAAAAAAAGTGGTTTATCGCTATCAACGCAACAGCGAATACGCCGACTTTTCGCAGAAAGAGATTTTTCACCTTAAAGGCTTCGGATTCACCGGGCTTGTAGGCCTGTCAATTTTTTCCACTTATTTAAATGTTTGTAAATTAAGTAAATAACGTTTTTAATCTAAAAATTCCACAATTTCCACAATAACTACTACTACTAAATTTTTTTAATTATTTTATTTATTTATATTAGAAAAGAGAACAATATAAGTTATTGAATTTAATTTACGAAAAAAGTATATTGTGTATGTGTTAAAAACTTCCAAATTCCGATGGAAATTATTTGTAATCAAAATGAGTTAAACAATGCTATACAACTAGTAAGTAAAGCAGTTGCTTCACGACCAACTCATCCAATTCTTGCAAATATACTTTTAACAGCTGACCAAGGAACTAATAAAATTAGTTTGACAGGATTTGATCTTAATTTAGGAATTCAAACTTCTTTTGATGGAACTGTTAAAAATAGCGGTGCTATTACGATACCTTCAAAACTTTTATCTGAAATAGTAAACAAATTAC...(this still goes for a lot more A or C or G or T)

>NC_010391.1 Salmonella phage Fels-1, complete genome 
TGATTGCATAACCACTTAACATCTTGTTTTATCTAAATAAAATTAAGCATGTTATCTTTTTGGGGCACTCCTGGGGCAGTAGATGCCAGTTGTTGATTCAGTATATCTACTTGTGACTGGTTATTATCCCGATTTTTTTAGTTTTAAGGTGTTGACATAGCCATCCATGCTCCATATACTGTATAGACCATCTGAGCGTTTGAGTGCCCCATTTGTGAAGCAATAAAGTTCGGGTTCGCGCCAGCGGCAAGCGCCCAGCATGCACCGATTTTTTTAGTTTTAAGGTGTTGACATTAGGTATGTCGGGACTGGTATGCTTTCCTGTGTCGCAGCCCGGCGCGTCTCAATGCAGATTCCCATATCCTGTTCAGGGAAGGTGCAGCATAATGTGCTTTGGATTTACCTGAGCGATCGGTAATTTGCGGATTGAAGACAAAGGTAATTTTTTCCACTTATTTAAATGTTTGTAAATTAAGTAAATAACGTTTTTAATCTAAAAATTCCACAATTTCCACAATAACTACTACTACTAAATTTTTTTAATTATTTTATTTATTTATATTAGAAAAGAGAACAATATAAGTTATTGAATTTAATTTACGAAAAAAGTATATTGTGTATGTGTTAAAAACTTCCAAATTCCGATGGAAATTATTTGTAATCAAAATGAGTTAAACAATGCTATACAACTAGTAAGTAAAGCAGTTGCTTCACGACCAACTCATCCAATTCTTGCAAATATACTTTTAACAGCTGACCAAGGAACTAATAAAATTAGTTTGACAGGATTTGATCTTAATTTAGGAATTCAAACTTCTTTTGATGGAACTGTTAAAAATAGCGGTGCTATTACGATACCTTCAAAACTTTTATCTGAAATAGTAAACAAATTAC...(this still goes for a lot more A or C or G or T)

>NC_007577.1 Prochlorococcus marinus str. MIT 9312, complete genome 
AATTTTTTCCACTTATTTAAATGTTTGTAAATTAAGTAAATAACGTTTTTAATCTAAAAATTCCACAATTTCCACAATAACTACTACTACTAAATTTTTTTAATTATTTTATTTATTTATATTAGAAAAGAGAACAATATAAGTTATTGAATTTAATTTACGAAAAAAGTATATTGTGTATGTGTTAAAAACTTCCAAATTCCGATGGAAATTATTTGTAATCAAAATGAGTTAAACAATGCTATACAACTAGTAAGTAAAGCAGTTGCTTCACGACCAACTCATCCAATTCTTGCAAATATACTTTTAACAGCTGACCAAGGAACTAATAAAATTAGTTTGACAGGATTTGATCTTAATTTAGGAATTCAAACTTCTTTTGATGGAACTGTTAAAAATAGCGGTGCTATTACGATACCTTCAAAACTTTTATCTGAAATAGTAAACAAATTACAATTTTTTCCACTTATTTAAATGTTTGTAAATTAAGTAAATAACGTTTTTAATCTAAAAATTCCACAATTTCCACAATAACTACTACTACTAAATTTTTTTAATTATTTTATTTATTTATATTAGAAAAGAGAACAATATAAGTTATTGAATTTAATTTACGAAAAAAGTATATTGTGTATGTGTTAAAAACTTCCAAATTCCGATGGAAATTATTTGTAATCAAAATGAGTTAAACAATGCTATACAACTAGTAAGTAAAGCAGTTGCTTCACGACCAACTCATCCAATTCTTGCAAATATACTTTTAACAGCTGACCAAGGAACTAATAAAATTAGTTTGACAGGATTTGATCTTAATTTAGGAATTCAAACTTCTTTTGATGGAACTGTTAAAAATAGCGGTGCTATTACGATACCTTCAAAACTTTTATCTGAAATAGTAAACAAATTAC

So for my new output, I will end up with a file with 10,000 subsets of 200 characters that would look like this:

>1
ATGGCAGATAAACGAATCCGTTCCGACAGTTCGGCGGCAGCGGTTCAGGCCATGAAAAATGCAGCTGTGGCTTCTCCCGCTTCAGTCTGCCAATATGGATGTGAAACTCGTCGGAAAAAAAGTGGTTTATCGCTATCAACGCAACAGCGAATACGCCGACTTTTTCAGTCTGCCAATATGCGTTCCGACAGCAGATAAAC

>2
TGATTGCATAACCACTTAACATCTTGTTTTATCTAAATAAAATTAAGCATGTTATCTTTTTGGGGCACTCCTGGGGCAGTAGATGCCAGTTGTTGATTCAGTATATCTACTTGTGACTGGTTATTATCCCGATTTTTTTAGTTTTAAGGTGTTGACATAGCCATCCATGCTCCATATACTGTATAGACCATCTGCAGATA

>3
AATTTTTTCCACTTATTTAAATGTTTGTAAATTAAGTAAATAACGTTTTTAATCTAAAAATTCCACAATTTCCACAATAACTACTACTACTAAATTTTTTTAATTATTTTATTTATTTATATTAGAAAAGAGAACAATATAAGTTATTGAATTTAATTTACGAAAAAAGTATATTGTGTATGTGTTAAAAACTTCCAAAT

Makes sense? Any help?

Thanks in advance

Answer 1

Given that you want to simulate short-read sequencing from reference genomes, there are read simulator programs that already exist. For example, see this answer.