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A route planner for DNA assembly

Home Page: https://dnaweaver.genomefoundry.org

License: MIT License

Python 97.06% HTML 1.92% CSS 1.02%

cloning-strategy dna-assembly planning synthetic-biology

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sandyg05 tschinz juliozohar andrewshvv jlerman44 alexsongrj asimovbio barionleg xiebaoping

dnaweaver's Issues

File Not Found Error during assembly_report execution

Hello! I discovered DNA Weaver quite recently and started diving into the documentation to learn how to use it.

The documentation and provided examples are quite useful, however I've found myself trapped into a situation that is not contemplated within the frequently asked questions or other support topics.

I think it could be useful to include this, and other common issues within a "Common Issues" section or similar.

The problem I am finding is the following:

Whenever I try to execute an script from the "tests" or "examples" folder, the script stops indicating the same error:

` File "C:\Users\fmjor\Desktop\DnaWeaver-master\examples\scenarios\example_with_adapters\example_with_adapters.py", line 97, in
assembly_report = quote.to_assembly_plan_report()

File "C:\ProgramData\Anaconda3\lib\site-packages\dnaweaver\DnaQuote\ExportsMixin.py", line 163, in to_assembly_plan_report
self.compute_fragments_final_locations()

File "C:\ProgramData\Anaconda3\lib\site-packages\dnaweaver\DnaQuote\PostProcessingMixin.py", line 64, in compute_fragments_final_locations
results = blast_sequence(

File "C:\ProgramData\Anaconda3\lib\site-packages\dnaweaver\biotools\sequence_homologies.py", line 82, in blast_sequence
p = subprocess.Popen(

File "C:\ProgramData\Anaconda3\lib\site-packages\spyder_kernels\customize\spydercustomize.py", line 105, in init
super(SubprocessPopen, self).init(*args, **kwargs)

File "C:\ProgramData\Anaconda3\lib\subprocess.py", line 854, in init
self._execute_child(args, executable, preexec_fn, close_fds,

File "C:\ProgramData\Anaconda3\lib\subprocess.py", line 1307, in _execute_child
hp, ht, pid, tid = _winapi.CreateProcess(executable, args,

FileNotFoundError: [WinError 2] The system can not found the specified file`

I guess this message is indicating that the program can not find the required files for BLAST and other operations. But I wonder if it could be possible to fix it.

Thanks i advance!

Bug: Max length initialisation

I believe it should be sequence_length + 1 otherwise GoldenGatePartLibrary might be ignored if the whole sequence is from the library.

DnaWeaver/dnaweaver/DnaSupplier/builtin_suppliers/DnaAssemblyStation/SequenceDecomposer.py

Line 124 in 6a26b08

self.max_segment_length = sequence_length

Because here you make a comparison of this form:

DnaWeaver/dnaweaver/DnaSupplier/builtin_suppliers/DnaAssemblyStation/SequenceDecomposer.py

Line 173 in 6a26b08

ends_max = min(L + 1, start + self.max_segment_length)

dealing with circular DNA

Hey-- this is an awesome library, and thanks for developing and sharing it.

I've been playing around with it to think about how I might be able to use it, and I've been testing it by creating a blast db with a bunch of preexisting vectors. I'm testing by trying to build a new one with a small insert, and it looks like the strategies are going to work. However, the origin of those vectors seems to be causing what I consider to be suboptimal solutions. All solutions are creating a junction at the origin with two PCR products rather than making one product going across the origin.

Is this a known issue? I've also tried making a Golden Gate-compatible version of that vector and loading it into a GoldenGatePartsLibrary, but I still just get the suboptimal PCR product solutions. Do you think I might be doing something wrong?

Q: Any specific reason why PcrExtractionStation not using the PartsLibrary?

Hey, I am using the PcrExtractionStation atm, and have noticed that it uses the blast database, I wonder, are there any architectural reasons why it is not using the PartsLibrary instead?

Such a decision leads to the fact that the PCR matrix does not appear in the assembly_parts field of the report.

Just feedback, maybe someone will be against it, but the ideal behavior from my POV would be to:

Upload seq records with annotations in PartLibrary.
Specify the same parts library for PCRExtractionStation as well as GoldenGate DNAAssemblyStation.
When PCRExtraction is used, the assembly plan contains a matrix seq record and synthesized primers.

Basically, internally in the PartsLibrary blast database is created and used for a search, and if the record is found, the seq record is used instead as an output. In this case, the annotation will be preserved along the way. Also in this case the cost function will be split - PCR extraction cost will include the cost of actual extraction, and PartLibrary might include the cost of delivery if some external provider is used.

On a side note: I am forced to use the blast database because when sequences are uploaded directly in the PCRExraction station it produces invalid PCR extraction operations. I might upload such a case soon.

SBOL => Automation MoClo Assembly tool => Optimal assembly report

Context
I am trying to create/use an automated design tool that will take the SBOL file of the desired construct and create an optimal way of assembling it using the MoClo standard.

After playing some time with DNAWeawer I believe I am not able to achieve that with this library, though I am still not 100% sure.

Question
Q: Have you seen libraries like that?

Bug: When quote.assembly_plan is None

When quote.assembly plan is None and the source quote returns an error quote, the assembly plan of the second quote is None, and an exception happens.

DnaWeaver/dnaweaver/DnaQuote/PostProcessingMixin.py

Line 32 in 6a26b08

quote.assembly_plan.items(), key=lambda item: item[0]

[WARNING][2021-05-06 21:51:29,214][__init__.py:261 - <module>()] gel simulation will NOT be available. Missing modules: mpldatacursor, pint
Traceback (most recent call last):
  File "/Users/Andrey/PycharmProjects/cloning-facility-core/src/order_parser/backend/decomposition/expression_vector.py", line 34, in decompose_expression_vector
    golden_gate_parts = decompose_insert(insert=insert,
  File "/Users/Andrey/PycharmProjects/cloning-facility-core/src/order_parser/backend/decomposition/synthesis.py", line 92, in decompose_insert
    assembly_plan_report = quote.to_assembly_plan_report()
  File "/Users/Andrey/PycharmProjects/cloning-facility-core/venv/lib/python3.9/site-packages/dnaweaver/DnaQuote/ExportsMixin.py", line 163, in to_assembly_plan_report
    self.compute_fragments_final_locations()
  File "/Users/Andrey/PycharmProjects/cloning-facility-core/venv/lib/python3.9/site-packages/dnaweaver/DnaQuote/PostProcessingMixin.py", line 57, in compute_fragments_final_locations
    self.compute_full_assembly_plan()
  File "/Users/Andrey/PycharmProjects/cloning-facility-core/venv/lib/python3.9/site-packages/dnaweaver/DnaQuote/PostProcessingMixin.py", line 43, in compute_full_assembly_plan
    rec(self)
  File "/Users/Andrey/PycharmProjects/cloning-facility-core/venv/lib/python3.9/site-packages/dnaweaver/DnaQuote/PostProcessingMixin.py", line 29, in rec
    segments = {
  File "/Users/Andrey/PycharmProjects/cloning-facility-core/venv/lib/python3.9/site-packages/dnaweaver/DnaQuote/PostProcessingMixin.py", line 30, in <dictcomp>
    segment: rec(subquote)
  File "/Users/Andrey/PycharmProjects/cloning-facility-core/venv/lib/python3.9/site-packages/dnaweaver/DnaQuote/PostProcessingMixin.py", line 32, in rec
    quote.assembly_plan.items(), key=lambda item: item[0]
AttributeError: 'NoneType' object has no attribute 'items

Contact Zulko

Hey @Zulko, what is the best way to contact u if I have questions regarding the DnaWeaver functionality?

verify_constraints performance

I have noticed that the verify_constraints is the most called function. It takes about 50% of execution time, which slows down the program and development cycle for real use cases (10kb sequences).

What is the proper way to implement the caching mechanism?

Does the PartsLibrary work for circular sequences?

Context

I have a setup of:

Golden gate parts library supplier with expression vectors.
Gene synthesis supplier.
Golden Gate assembly station.

I have uploaded the backbone sequence to the GoldenGatePartsLibrary, expecting DnaWeawer to find it.

I am giving the library a final sequence of expression vector with inserted gene as input.

Expected behavior:

Gene being synthesized
Backbone being taken from the parts library

Actual behaviour:

Everything is synthesised and assembled with GoldenGate

Question:

Q: Does the PartsLibrary work for circular sequences?

Example of using PartsLibrary with Gibson assembly method

I want to make a PartsLibrary that consists of basic stock PCR products that will be assembled with a DnaAssemblyStation based on the GibsonAssemblyMethod and any missing parts should come from a CommercialDnaOffer.

I have tried to make my own PartsLibrary class which inherets from the PartsLibrary base class, but am not able to make it work.

Do you have any examples availible of using a PartsLibrary that does not consists of GoldenGate parts, which is all that i can find in the documentation?

What role cut_spread_radius plays in circular sequence decomposition?

I am debugging why the circular sequence decomposition does not work for me, and it seems cut_spread_radius kinda plays a role in it. The documentation is lacking information about it.

Could you please share a bit of information about it?

What to do when Gibson assembly Station is refused : Apologies to put a routine use question as an issue

Hi All I am a newbie to DNAWeaver and want to use it to optimize my multipart Gibson assembly for ~8Kb plasmids

My code is given below and the error I get is

From Gibson Assembly Station - refused: No solution found! One forced cut was also invalid at indices {8851} - No solution found! One forced cut was also invalid at indices {8851}

I have tried changing the Gibson assembly station parameters and don't get anything other than the message above. How do I troubleshoot this ? Apologies if I am not understanding some basic fact about DNAweaver. I can run the examples perfectly but cant get this or other sequences to give me a result.

Thanks

import dnaweaver as dw

twist_dnafragments_offer = dw.CommercialDnaOffer(
    name="TwistDNAFragments",
    sequence_constraints=[
        dw.SequenceLengthConstraint(min_length=300,max_length=1800)
    ],
    pricing=dw.PerBasepairPricing(0.07),
    lead_time=40
)

twist_dnafclonalgenes_offer = dw.CommercialDnaOffer(
    name="TwistDNAClonalGenes",
    sequence_constraints=[
        dw.SequenceLengthConstraint(min_length=300,max_length=5000)
    ],
    pricing=dw.PerBasepairPricing(0.09),
    lead_time=40
)

assembly_station = dw.DnaAssemblyStation(
    name="Gibson Assembly Station",
    assembly_method=dw.GibsonAssemblyMethod(
        overhang_selector=dw.TmSegmentSelector(min_tm=55, max_tm=65),
        min_segment_length=1000,
        max_segment_length=4000,
        duration=5
    ),
    supplier=[twist_dnafragments_offer,twist_dnafclonalgenes_offer],
    coarse_grain=20
)
# sequence = dw.random_dna_sequence(8851, seed=123)
sequence = """CAGTACTCGTGCATAACAATCGCGTAATCGGCGAAGGTTGGAATAGGCCGATCGGACGCCACGACCCCAC
TGCACATGCGGAAATCATGGCCCTTCGACAGGGAGGGCTTGTGATGCAGAATTATCGACTTATCGATGCG
ACGCTGTACGTCACGCTTGAACCTTGCGTAATGTGCGCGGGAGCTATGATTCACTCCCGCATTGGACGAG
TTGTATTCGGTGCCCGCGACGCCAAGACGGGTGCCGCAGGTTCACTGATGGACGTGCTGCATCACCCAGG
CATGAACCACCGGGTAGAAATCACAGAAGGCATATTGGCGGACGAATGTGCGGCGCTGTTGTCCGACTTT
TTTCGCATGCGGAGGCAGGAGATCAAGGCCCAGAAAAAAGCACAATCCTCTACTGACTCTGGTGGTTCTT
CTGGTGGTTCTAGCGGCAGCGAGACTCCCGGGACCTCAGAGTCCGCCACACCCGAAAGTTCTGGTGGTTC
TTCTGGTGGTTCTTCCGAAGTCGAGTTTTCCCATGAGTACTGGATGAGACACGCATTGACTCTCGCAAAG
AGGGCTCGAGATGAACGCGAGGTGCCCGTGGGGGCAGTACTCGTGCTCAACAATCGCGTAATCGGCGAAG
GTTGGAATAGGGCAATCGGACTCCACGACCCCACTGCACATGCGGAAATCATGGCCCTTCGACAGGGAGG
GCTTGTGATGCAGAATTATCGACTTATCGATGCGACGCTGTACGTCACGTTTGAACCTTGCGTAATGTGC
GCGGGAGCTATGATTCACTCCCGCATTGGACGAGTTGTATTCGGTGTTCGCAACGCCAAGACGGGTGCCG
CAGGTTCACTGATGGACGTGCTGCATTACCCAGGCATGAACCACCGGGTAGAAATCACAGAAGGCATATT
GGCGGACGAATGTGCGGCGCTGTTGTGTTACTTTTTTCGCATGCCCAGGCAGGTCTTTAACGCCCAGAAA
AAAGCACAATCCTCTACTGACTCTGGTGGTTCTTCTGGTGGTTCTAGCGGCAGCGAGACTCCCGGGACCT
CAGAGTCCGCCACACCCGAAAGTTCTGGTGGTTCTTCTGGTGGTTCTGATAAAAAGTATTCTATTGGTTT
AGCCATCGGCACTAATTCCGTTGGATGGGCTGTCATAACCGATGAATACAAAGTACCTTCAAAGAAATTT
AAGGTGTTGGGGAACACAGACCGTCATTCGATTAAAAAGAATCTTATCGGTGCCCTCCTATTCGATAGTG
GCGAAACGGCAGAGGCGACTCGCCTGAAACGAACCGCTCGGAGAAGGTATACACGTCGCAAGAACCGAAT
ATGTTACTTACAAGAAATTTTTAGCAATGAGATGGCCAAAGTTGACGATTCTTTCTTTCACCGTTTGGAA
GAGTCCTTCCTTGTCGAAGAGGACAAGAAACATGAACGGCACCCCATCTTTGGAAACATAGTAGATGAGG
TGGCATATCATGAAAAGTACCCAACGATTTATCACCTCAGAAAAAAGCTAGTTGACTCAACTGATAAAGC
GGACCTGAGGTTAATCTACTTGGCTCTTGCCCATATGATAAAGTTCCGTGGGCACTTTCTCATTGAGGGT
GATCTAAATCCGGACAACTCGGATGTCGACAAACTGTTCATCCAGTTAGTACAAACCTATAATCAGTTGT
TTGAAGAGAACCCTATAAATGCAAGTGGCGTGGATGCGAAGGCTATTCTTAGCGCCCGCCTCTCTAAATC
CCGACGGCTAGAAAACCTGATCGCACAATTACCCGGAGAGAAGAAAAATGGGTTGTTCGGTAACCTTATA
GCGCTCTCACTAGGCCTGACACCAAATTTTAAGTCGAACTTCGACTTAGCTGAAGATGCCAAATTGCAGC
TTAGTAAGGACACGTACGATGACGATCTCGACAATCTACTGGCACAAATTGGAGATCAGTATGCGGACTT
ATTTTTGGCTGCCAAAAACCTTAGCGATGCAATCCTCCTATCTGACATACTGAGAGTTAATACTGAGATT
ACCAAGGCGCCGTTATCCGCTTCAATGATCAAAAGGTACGATGAACATCACCAAGACTTGACACTTCTCA
AGGCCCTAGTCCGTCAGCAACTGCCTGAGAAATATAAGGAAATATTCTTTGATCAGTCGAAAAACGGGTA
CGCAGGTTATATTGACGGCGGAGCGAGTCAAGAGGAATTCTACAAGTTTATCAAACCCATATTAGAGAAG
ATGGATGGGACGGAAGAGTTGCTTGTAAAACTCAATCGCGAAGATCTACTGCGAAAGCAGCGGACTTTCG
ACAACGGTAGCATTCCACATCAAATCCACTTAGGCGAATTGCATGCTATACTTAGAAGGCAGGAGGATTT
TTATCCGTTCCTCAAAGACAATCGTGAAAAGATTGAGAAAATCCTAACCTTTCGCATACCTTACTATGTG
GGACCCCTGGCCCGAGGGAACTCTCGGTTCGCATGGATGACAAGAAAGTCCGAAGAAACGATTACTCCAT
GGAATTTTGAGGAAGTTGTCGATAAAGGTGCGTCAGCTCAATCGTTCATCGAGAGGATGACCAACTTTGA
CAAGAATTTACCGAACGAAAAAGTATTGCCTAAGCACAGTTTACTTTACGAGTATTTCACAGTGTACAAT
GAACTCACGAAAGTTAAGTATGTCACTGAGGGCATGCGTAAACCCGCCTTTCTAAGCGGAGAACAGAAGA
AAGCAATAGTAGATCTGTTATTCAAGACCAACCGCAAAGTGACAGTTAAGCAATTGAAAGAGGACTACTT
TAAGAAAATTGAATGCTTCGATTCTGTCGAGATCTCCGGGGTAGAAGATCGATTTAATGCGTCACTTGGT
ACGTATCATGACCTCCTAAAGATAATTAAAGATAAGGACTTCCTGGATAACGAAGAGAATGAAGATATCT
TAGAAGATATAGTGTTGACTCTTACCCTCTTTGAAGATCGGGAAATGATTGAGGAAAGACTAAAAACATA
CGCTCACCTGTTCGACGATAAGGTTATGAAACAGTTAAAGAGGCGTCGCTATACGGGCTGGGGACGATTG
TCGCGGAAACTTATCAACGGGATAAGAGACAAGCAAAGTGGTAAAACTATTCTCGATTTTCTAAAGAGCG
ACGGCTTCGCCAATAGGAACTTTATGCAGCTGATCCATGATGACTCTTTAACCTTCAAAGAGGATATACA
AAAGGCACAGGTTTCCGGACAAGGGGACTCATTGCACGAACATATTGCGAATCTTGCTGGTTCGCCAGCC
ATCAAAAAGGGCATACTCCAGACAGTCAAAGTAGTGGATGAGCTAGTTAAGGTCATGGGACGTCACAAAC
CGGAAAACATTGTAATCGAGATGGCACGCGAAAATCAAACGACTCAGAAGGGGCAAAAAAACAGTCGAGA
GCGGATGAAGAGAATAGAAGAGGGTATTAAAGAACTGGGCAGCCAGATCTTAAAGGAGCATCCTGTGGAA
AATACCCAATTGCAGAACGAGAAACTTTACCTCTATTACCTACAAAATGGAAGGGACATGTATGTTGATC
AGGAACTGGACATAAACCGTTTATCTGATTACGACGTCGATCACATTGTACCCCAATCCTTTTTGAAGGA
CGATTCAATCGACAATAAAGTGCTTACACGCTCGGATAAGAACCGAGGGAAAAGTGACAATGTTCCAAGC
GAGGAAGTCGTAAAGAAAATGAAGAACTATTGGCGGCAGCTCCTAAATGCGAAACTGATAACGCAAAGAA
AGTTCGATAACTTAACTAAAGCTGAGAGGGGTGGCTTGTCTGAACTTGACAAGGCCGGATTTATTAAACG
TCAGCTCGTGGAAACCCGCCAAATCACAAAGCATGTTGCACAGATACTAGATTCCCGAATGAATACGAAA
TACGACGAGAACGATAAGCTGATTCGGGAAGTCAAAGTAATCACTTTAAAGTCAAAATTGGTGTCGGACT
TCAGAAAGGATTTTCAATTCTATAAAGTTAGGGAGATAAATAACTACCACCATGCGCACGACGCTTATCT
TAATGCCGTCGTAGGGACCGCACTCATTAAGAAATACCCGAAGCTAGAAAGTGAGTTTGTGTATGGTGAT
TACAAAGTTTATGACGTCCGTAAGATGATCGCGAAAAGCGAACAGGAGATAGGCAAGGCTACAGCCAAAT
ACTTCTTTTATTCTAACATTATGAATTTCTTTAAGACGGAAATCACTCTGGCAAACGGAGAGATACGCAA
ACGACCTTTAATTGAAACCAATGGGGAGACAGGTGAAATCGTATGGGATAAGGGCCGGGACTTCGCGACG
GTGAGAAAAGTTTTGTCCATGCCCCAAGTCAACATAGTAAAGAAAACTGAGGTGCAGACCGGAGGGTTTT
CAAAGGAATCGATTCTTCCAAAAAGGAATAGTGATAAGCTCATCGCTCGTAAAAAGGACTGGGACCCGAA
AAAGTACGGTGGCTTCGATAGCCCTACAGTTGCCTATTCTGTCCTAGTAGTGGCAAAAGTTGAGAAGGGA
AAATCCAAGAAACTGAAGTCAGTCAAAGAATTATTGGGGATAACGATTATGGAGCGCTCGTCTTTTGAAA
AGAACCCCATCGACTTCCTTGAGGCGAAAGGTTACAAGGAAGTAAAAAAGGATCTCATAATTAAACTACC
AAAGTATAGTCTGTTTGAGTTAGAAAATGGCCGAAAACGGATGTTGGCTAGCGCCGGAGAGCTTCAAAAG
GGGAACGAACTCGCACTACCGTCTAAATACGTGAATTTCCTGTATTTAGCGTCCCATTACGAGAAGTTGA
AAGGTTCACCTGAAGATAACGAACAGAAGCAACTTTTTGTTGAGCAGCACAAACATTATCTCGACGAAAT
CATAGAGCAAATTTCGGAATTCAGTAAGAGAGTCATCCTAGCTGATGCCAATCTGGACAAAGTATTAAGC
GCATACAACAAGCACAGGGATAAACCCATACGTGAGCAGGCGGAAAATATTATCCATTTGTTTACTCTTA
CCAACCTCGGCGCTCCAGCCGCATTCAAGTATTTTGACACAACGATAGATCGCAAACGATACACTTCTAC
CAAGGAGGTGCTAGACGCGACACTGATTCACCAATCCATCACGGGATTATATGAAACTCGGATAGATTTG
TCACAGCTTGGGGGTGACTCTGGTGGTTCTCCCAAGAAGAAGAGGAAAGTCTAACCGGTCATCATCACCA
TCACCATTGAGTTTAAACCCGCTGATCAGCCTCGACTGTGCCTTCTAGTTGCCAGCCATCTGTTGTTTGC
CCCTCCCCCGTGCCTTCCTTGACCCTGGAAGGTGCCACTCCCACTGTCCTTTCCTAATAAAATGAGGAAA
TTGCATCGCATTGTCTGAGTAGGTGTCATTCTATTCTGGGGGGTGGGGTGGGGCAGGACAGCAAGGGGGA
GGATTGGGAAGACAATAGCAGGCATGCTGGGGATGCGGTGGGCTCTATGGCTTCTGAGGCGGAAAGAACC
AGCTGGGGCTCGATACCGTCGACCTCTAGCTAGAGCTTGGCGTAATCATGGTCATAGCTGTTTCCTGTGT
GAAATTGTTATCCGCTCACAATTCCACACAACATACGAGCCGGAAGCATAAAGTGTAAAGCCTAGGGTGC
CTAATGAGTGAGCTAACTCACATTAATTGCGTTGCGCTCACTGCCCGCTTTCCAGTCGGGAAACCTGTCG
TGCCAGCTGCATTAATGAATCGGCCAACGCGCGGGGAGAGGCGGTTTGCGTATTGGGCGCTCTTCCGCTT
CCTCGCTCACTGACTCGCTGCGCTCGGTCGTTCGGCTGCGGCGAGCGGTATCAGCTCACTCAAAGGCGGT
AATACGGTTATCCACAGAATCAGGGGATAACGCAGGAAAGAACATGTGAGCAAAAGGCCAGCAAAAGGCC
AGGAACCGTAAAAAGGCCGCGTTGCTGGCGTTTTTCCATAGGCTCCGCCCCCCTGACGAGCATCACAAAA
ATCGACGCTCAAGTCAGAGGTGGCGAAACCCGACAGGACTATAAAGATACCAGGCGTTTCCCCCTGGAAG
CTCCCTCGTGCGCTCTCCTGTTCCGACCCTGCCGCTTACCGGATACCTGTCCGCCTTTCTCCCTTCGGGA
AGCGTGGCGCTTTCTCATAGCTCACGCTGTAGGTATCTCAGTTCGGTGTAGGTCGTTCGCTCCAAGCTGG
GCTGTGTGCACGAACCCCCCGTTCAGCCCGACCGCTGCGCCTTATCCGGTAACTATCGTCTTGAGTCCAA
CCCGGTAAGACACGACTTATCGCCACTGGCAGCAGCCACTGGTAACAGGATTAGCAGAGCGAGGTATGTA
GGCGGTGCTACAGAGTTCTTGAAGTGGTGGCCTAACTACGGCTACACTAGAAGAACAGTATTTGGTATCT
GCGCTCTGCTGAAGCCAGTTACCTTCGGAAAAAGAGTTGGTAGCTCTTGATCCGGCAAACAAACCACCGC
TGGTAGCGGTGGTTTTTTTGTTTGCAAGCAGCAGATTACGCGCAGAAAAAAAGGATCTCAAGAAGATCCT
TTGATCTTTTCTACGGGGTCTGACGCTCAGTGGAACGAAAACTCACGTTAAGGGATTTTGGTCATGAGAT
TATCAAAAAGGATCTTCACCTAGATCCTTTTAAATTAAAAATGAAGTTTTAAATCAATCTAAAGTATATA
TGAGTAAACTTGGTCTGACAGTTACCAATGCTTAATCAGTGAGGCACCTATCTCAGCGATCTGTCTATTT
CGTTCATCCATAGTTGCCTGACTCCCCGTCGTGTAGATAACTACGATACGGGAGGGCTTACCATCTGGCC
CCAGTGCTGCAATGATACCGCGAGACCCACGCTCACCGGCTCCAGATTTATCAGCAATAAACCAGCCAGC
CGGAAGGGCCGAGCGCAGAAGTGGTCCTGCAACTTTATCCGCCTCCATCCAGTCTATTAATTGTTGCCGG
GAAGCTAGAGTAAGTAGTTCGCCAGTTAATAGTTTGCGCAACGTTGTTGCCATTGCTACAGGCATCGTGG
TGTCACGCTCGTCGTTTGGTATGGCTTCATTCAGCTCCGGTTCCCAACGATCAAGGCGAGTTACATGATC
CCCCATGTTGTGCAAAAAAGCGGTTAGCTCCTTCGGTCCTCCGATCGTTGTCAGAAGTAAGTTGGCCGCA
GTGTTATCACTCATGGTTATGGCAGCACTGCATAATTCTCTTACTGTCATGCCATCCGTAAGATGCTTTT
CTGTGACTGGTGAGTACTCAACCAAGTCATTCTGAGAATAGTGTATGCGGCGACCGAGTTGCTCTTGCCC
GGCGTCAATACGGGATAATACCGCGCCACATAGCAGAACTTTAAAAGTGCTCATCATTGGAAAACGTTCT
TCGGGGCGAAAACTCTCAAGGATCTTACCGCTGTTGAGATCCAGTTCGATGTAACCCACTCGTGCACCCA
ACTGATCTTCAGCATCTTTTACTTTCACCAGCGTTTCTGGGTGAGCAAAAACAGGAAGGCAAAATGCCGC
AAAAAAGGGAATAAGGGCGACACGGAAATGTTGAATACTCATACTCTTCCTTTTTCAATATTATTGAAGC
ATTTATCAGGGTTATTGTCTCATGAGCGGATACATATTTGAATGTATTTAGAAAAATAAACAAATAGGGG
TTCCGCGCACATTTCCCCGAAAAGTGCCACCTGACGTCGACGGATCGGGAGATCGATCTCCCGATCCCCT
AGGGTCGACTCTCAGTACAATCTGCTCTGATGCCGCATAGTTAAGCCAGTATCTGCTCCCTGCTTGTGTG
TTGGAGGTCGCTGAGTAGTGCGCGAGCAAAATTTAAGCTACAACAAGGCAAGGCTTGACCGACAATTGCA
TGAAGAATCTGCTTAGGGTTAGGCGTTTTGCGCTGCTTCGCGATGTACGGGCCAGATATACGCGTTGACA
TTGATTATTGACTAGTTATTAATAGTAATCAATTACGGGGTCATTAGTTCATAGCCCATATATGGAGTTC
CGCGTTACATAACTTACGGTAAATGGCCCGCCTGGCTGACCGCCCAACGACCCCCGCCCATTGACGTCAA
TAATGACGTATGTTCCCATAGTAACGCCAATAGGGACTTTCCATTGACGTCAATGGGTGGAGTATTTACG
GTAAACTGCCCACTTGGCAGTACATCAAGTGTATCATATGCCAAGTACGCCCCCTATTGACGTCAATGAC
GGTAAATGGCCCGCCTGGCATTATGCCCAGTACATGACCTTATGGGACTTTCCTACTTGGCAGTACATCT
ACGTATTAGTCATCGCTATTACCATGGTGATGCGGTTTTGGCAGTACATCAATGGGCGTGGATAGCGGTT
TGACTCACGGGGATTTCCAAGTCTCCACCCCATTGACGTCAATGGGAGTTTGTTTTGGCACCAAAATCAA
CGGGACTTTCCAAAATGTCGTAACAACTCCGCCCCATTGACGCAAATGGGCGGTAGGCGTGTACGGTGGG
AGGTCTATATAAGCAGAGCTGGTTTAGTGAACCGTCAGATCCGCTAGAGATCCGCGGCCGCTAATACGAC
TCACTATAGGGAGAGCCGCCACCATGTCCGAAGTCGAGTTTTCCCATGAGTACTGGATGAGACACGCATT
GACTCTCGCAAAGAGGGCTTGGGATGAACGCGAGGTGCCCGTGGGGG"""
quote = assembly_station.get_quote(sequence, with_assembly_plan=True)

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