View source for Talk:Biosynthesis of DMT in modified organisms using genetic engineering

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[[File:Fc906056bb7f24546fd8af645ad4c08d.jpg|thumb]]

The use of [[Genetic engineering]] to create the [[Biosynthesis]] of [[DMT]] in modified organisms

==Introduction==
As of June 2017 nobody has ever made a modification in a organism to produce DMT. 

This article contemplate the research that is going on in this subject. 

===Simplified steps to achieve the goal ===
# Learn about the genes involved  Biosynthesis of DMT in a specific organism that produce it like humans or Mimosa hostilis.
# Choose a organism that do not create DMT naturally like E.Coli or a specific plant.
# Write the plasmid digitally 
# Send the plasmid info (digital) to any place that sell plasmid and Buy the sequence that you want 
# Introduce the plasmid into the organisms (at home or pay a laboratoy) 
# Grow organisms that are producing DMT
# Harvest dmt using some Tek

==The theory - Research on how to do the gene modification==


===Biosynthesis of DMT in humans===
Dimethyltryptamine is an [[indole alkaloid]] derived from the [[shikimate]] pathway. 

the [[biosynthesis]] of DMT is relatively simple. 
[[Image:DMT biosynthetic pathway.png|thumb|center|Biosynthetic pathway for ''N'',''N''-dimethyltryptamine]]
[[L-tryptophan]] that we cosume is transformed in DMT in 4 step listed below. 

Humans cannot produce [[L-tryptophan]], so we need to eat it.
Tryptophan is a routine constituent of most protein-based foods or dietary proteins.  It is particularly plentiful in  [[milk]], [[yogurt]], [[cottage cheese]], [[red meat]], [[egg (food)|eggs]], etc.
 
[[File:L-Tryptophan - L-Tryptophan.svg|thumb|center|L-Tryptophan]]
It begins with its [[decarboxylation]] (a chemical reaction that removes a carboxyl group and releases carbon dioxide (CO2)) of [[L-tryptophan]] by an [[aromatic amino acid decarboxylase]] (AADC) [[enzymes|enzyme]] 
[[File:DOPA decarboxylase dimer 1JS3.png|thumb||center|Aromatic L-amino acid decarboxylase (AADC) ]]
(step 1). The resulting decarboxylated tryptophan [[Analog (chemistry)|analog]] is [[tryptamine]]. 

[[File:Tryptamine structure.svg|thumb|center|tryptamine ]]
Tryptamine then undergoes a [[transmethylation]] 

(step 2): the enzyme [[tryptamine-N-methyltransferase|indolethylamine-N-methyltransferase]] (INMT) [[Catalysis|catalyzes]] the transfer of a [[methyl group]] from [[Cofactor (biochemistry)|cofactor]] [[S-adenosyl-methionine]] (SAM), via [[nucleophilic]] attack, to tryptamine.
[[File:INMT.jpeg|thumb|center|Amine N-methyltransferase (INMT) ]]
This reaction transforms SAM into [[S-adenosylhomocysteine]] (SAH), and gives the intermediate product [[N-methyltryptamine|''N''-methyltryptamine]] (NMT).
[[File:Methyltryptamine.png|thumb|center|N-Methyltryptamine (NMT)  has been found in the bark, shoots and leaves of several plant genera, including Virola, Acacia, Mimosa]]
<ref name="pmid13685339">{{cite journal |author=Axelrod J. |title=Enzymatic formation of psychotomimetic metabolites from normally occurring compounds |journal=Science |volume=134 |issue= 3475|page=343 |date=August 1961 |pmid=13685339 |doi=10.1126/science.134.3475.343 |url=}}</ref><ref name="pmid779022">{{cite journal |author1=Rosengarten H. |author2=Friedhoff A.J. |title=A review of recent studies of the biosynthesis and excretion of hallucinogens formed by methylation of neurotransmitters or related substances |journal=Schizophrenia Bulletin |volume=2 |issue=1 |pages=90–105 |year=1976 |pmid=779022 |doi=10.1093/schbul/2.1.90 |url=http://schizophreniabulletin.oxfordjournals.org/content/2/1/90.full.pdf |format= PDF}}</ref> NMT is in turn transmethylated by the same process 

(step 3) to form the end product ''N'',''N''-dimethyltryptamine. Tryptamine transmethylation is regulated by two products of the reaction: SAH,<ref name="pmid6792104">{{cite journal |author1=Barker S.A. |author2=Monti J.A. |author3=Christian S.T. |title=''N, N''-dimethyltryptamine: an endogenous hallucinogen |journal=International Review of Neurobiology |volume=22 |issue= |pages=83–110 |year=1981 |pmid=6792104 |doi=10.1016/S0074-7742(08)60291-3 |series=International Review of Neurobiology |isbn=978-0-12-366822-6}}</ref><ref name="pmid4756800">{{cite journal |author1=Lin R.L. |author2=Narasimhachari N. |author3=Himwich H.E. |title=Inhibition of indolethylamine-N-methyltransferase by S-adenosylhomocysteine |journal=Biochemical and Biophysical Research Communications |volume=54 |issue=2 |pages=751–9 |date=September 1973 |pmid=4756800 |doi=10.1016/0006-291X(73)91487-3 |url=}}</ref><ref name="pmid9852119">{{cite journal |author1=Thompson M.A. |author2=Weinshilboum R.M. |title=Rabbit lung indolethylamine ''N''-methyltransferase. cDNA and gene cloning and characterization |journal=Journal of Biological Chemistry |volume=273 |issue=51 |pages=34502–10 |date=December 1998 |pmid=9852119 |doi=10.1074/jbc.273.51.34502 |url=http://www.jbc.org/content/273/51/34502.long |accessdate=2010-11-09}}</ref> and DMT<ref name="pmid6792104"/><ref name="pmid9852119"/> were shown ''ex vivo'' to be among the most potent inhibitors of rabbit INMT activity.

This transmethylation mechanism has been repeatedly and consistently proven by [[Isotope labeling|radiolabeling]] of SAM methyl group with [[carbon-14]] (<sup>14</sup>C-CH<sub>3</sub>)SAM).<ref name="pmid13685339"/><ref name="pmid6792104"/><ref name="pmid9852119"/><ref name="pmid14361">{{cite journal |author1=Mandel L.R. |author2=Prasad R. |author3=Lopez-Ramos B. |author4=Walker R.W. |title=The biosynthesis of dimethyltryptamine in vivo |journal=Research Communications in Chemical Pathology and Pharmacology |volume=16 |issue=1 |pages=47–58 |date=January 1977 |pmid=14361}}</ref><ref name="pmid10552930">{{cite journal |author1=Thompson M.A. |author2=Moon E. |author3=Kim U.J. |author4=Xu J. |author5=Siciliano M.J. |author6=Weinshilboum R.M. |title=Human indolethylamine ''N''-methyltransferase: cDNA cloning and expression, gene cloning, and chromosomal localization |journal=Genomics |volume=61 |issue=3 |pages=285–97 |date=November 1999 |pmid=10552930 |doi=10.1006/geno.1999.5960 |url=http://crfdl.org:1111/xmlui/bitstream/handle/123456789/307/Thompson99humanINMT.pdf?sequence=1 |format=PDF}}</ref>

===DMT and the human DNA===
Dmt in humans is created with genes located in Chromosome 7.

the genes are listed below

====[[Amine N-methyltransferase]]====
[[File:INMT.jpeg|thumb|Amine N-methyltransferase (INMT)]]

 EntrezGene = 21743
 UniProt = P40936
*located in [[Chromosome 7 (human)]] https://www.ncbi.nlm.nih.gov/genome/gdv/browser/?context=gene&acc=11185
{{enzyme
| Name = amine N-methyltransferase
| EC_number = 2.1.1.49
| CAS_number = 51377-47-0
| IUBMB_EC_number = 2/1/1/49
| GO_code = 0030748
| image = INMT.jpeg 
| width = 
| caption = indolethylamine N-methyltransferase (with slight variation on CPK coloration) – See PDB {{PDB2|2A14}} 
}}

In [[enzymology]], an [[Amine N-methyltransferase]] ({{EC number|2.1.1.49}}) is an enzyme that is ubiquitously present in non-neural tissues and that [[catalysis|catalyzes]] the N-[[methylation]] of [[tryptamine]] and structurally related compounds.<ref name="MeshName">{{MeshName|tryptamine+N-methyltransferase}}</ref>

The chemical reaction taking place is:
*[[S-adenosyl-L-methionine]] + an [[amine]] <math>\rightleftharpoons</math> [[S-adenosyl-L-homocysteine]] + a [[methylated]] amine

Thus, the two [[substrate (biochemistry)|substrates]] of this enzyme are [[S-Adenosyl methionine|S-adenosyl methionine]] and [[amine]], whereas its two [[product (chemistry)|products]] are [[S-adenosylhomocysteine]] and [[methylated amine]]. In the case of tryptamine and [[serotonin]] these then become the dimethylated indolethylamines [[dimethyltryptamine]] (DMT) and [[bufotenine]].<ref>{{cite journal |last= J. |first= Kärkkäinen |author2=T. Forsström|author3= J. Tornaeus|author4= K. Wähälä|author5= P. Kiuru|author6= A. Honkanen|author7= U. -H. Stenman|author8= U. Turpeinen|author9= A. Hesso |date=April 2005 |title= Potentially hallucinogenic 5-hydroxytryptamine receptor ligands bufotenine and dimethyltryptamine in blood and tissues |journal= Scandinavian Journal of Clinical and Laboratory Investigation |volume= 65 |issue= 3 |pages= 189–199 |id= |url= http://www.informaworld.com/smpp/content~content=a713735722~db=all~order=page |accessdate= October 15, 2008 |quote= |doi= 10.1080/00365510510013604 |pmid= 16095048  }}</ref>

This enzyme belongs to the family of [[transferase]]s, specifically those transferring one-carbon group methyltransferases.  The [[List of enzymes|systematic name]] of this enzyme class is '''S-adenosyl-L-methionine:amine N-methyltransferase'''. Other names in common use include '''nicotine N-methyltransferase''', '''tryptamine N-methyltransferase''', '''indolethylamine N-methyltransferase''', and '''arylamine N-methyltransferase'''.  This enzyme participates in [[tryptophan metabolism]].

A wide range of primary, secondary and tertiary amines can act as [[Electron acceptor|acceptors]], including tryptamine, [[aniline]], [[nicotine]] and a variety of drugs and other [[xenobiotic]]s.<ref name="MeshName"/>


N-methylation of endogenous and xenobiotic compounds is a major method by which they are degraded. This gene encodes an enzyme that N-methylates indoles such as tryptamine. Alternative splicing results in multiple transcript variants. Read-through transcription also exists between this gene and the downstream FAM188B (family with sequence similarity 188, member B) gene. [provided by RefSeq, Nov 2010]

Enzyme encoded into DNA

 atggaggggaaagtctatatcgggggggagttctatgagaaagagttcacacccaaattc tatctgacaacatattatagcttccatagcgggcccgtcgccgagcaagagatcgtcaaa 
 ttcagcctgcaaaatctgtatcaaacattcagcacagggggggtcggggggttcgtcctg atcttcatcgggagcgggcccacaatctatcaactgctgagcgcctgcgaggtcttcaga 
 gagatcatcgtcacattctatacaccccaaaatctgcaagagctgcaaaaatggctgaaa aaagagcccggggcctatttctggagcagcatcgtccaacatgcctgcgagctggagggg 
 ttcagaagcagatggcaagagaaagaggccaaactgagaagaacagtcacaagagtcctg agatgcttcgtcacaaaaacaccccccctggggagcgcccaagtccccctggccttctgc 
 gtcctgacattcctggccatggagtgcgcctgccccttcatcttcacatatagagccgcc ctgagaagactggccgggctgctgaaacccggggggcatctggtcacactggtcacactg 
 agattccaacattatatggtcgggcccaaaaaattcagcggggtctatctggagaaagag 
 gtcgtcgagaaagccatccaattcgccgggtgccaagtcctgaaatgcaattgcgtcagc  
 ctgagctatagcgaggcctattgcagccatttcgggctgtgcttcgtcgtcgccagaaaa 
 gggcccagc

DNA above is two steps away from the translation done by the ribosome. 
DNA is transcribed into RNA by DNA transcriptase, RNA is translated into proteins by ribosomes. 
But we are interested in the DNA that we can put into the organism, so that is why I worked my way backwards to DNA.

====[[Aromatic L-amino acid decarboxylase]]====
[[File:DOPA decarboxylase dimer 1JS3.png|thumb|Aromatic L-amino acid decarboxylase (AADC or AAAD)]]
http://www.uniprot.org/uniprot/O88533
 
{{enzyme
| Name = aromatic-L-amino-acid decarboxylase
| EC_number = 4.1.1.28
| CAS_number = 9042-64-2 
| IUBMB_EC_number = 4/1/1/28
| GO_code = 0004058
| image = DOPA decarboxylase dimer 1JS3.png
| width = 
| caption = Ribbon diagram of a DOPA decarboxylase [[protein dimer|dimer]].<ref name="pmid11685243">{{PDB|1JS3}}; {{cite journal | vauthors = Burkhard P, Dominici P, Borri-Voltattorni C, Jansonius JN, Malashkevich VN | title = Structural insight into Parkinson's disease treatment from drug-inhibited DOPA decarboxylase | journal = Nature Structural Biology | volume = 8 | issue = 11 | pages = 963–7 | date = Nov 2001 | pmid = 11685243 | doi = 10.1038/nsb1101-963 }}</ref>
}}
{{infobox protein
| Name = DOPA decarboxylase (aromatic <small>L</small>-amino acid decarboxylase)
| caption = 
| image = 
| width = 250
| HGNCid = 2719
| Symbol = DDC
| AltSymbols =
| EntrezGene = 1644
| OMIM = 107930
| RefSeq = NM_000790
| UniProt = P20711
| PDB =
| ECnumber = 4.1.1.28
| Chromosome = 7
| Arm = p
| Band = 11
| LocusSupplementaryData =
}}
[[Aromatic L-amino acid decarboxylase]] ('''AADC''' or '''AAAD'''), also known as '''DOPA decarboxylase''', '''tryptophan decarboxylase''', and '''5-hydroxytryptophan decarboxylase''', is a [[lyase]] [[enzyme]] ({{EC number|4.1.1.28}}).

=====Reactions=====
AADC catalyzes several different [[decarboxylation]] reactions:<ref>{{cite web|title=AADC|url=http://www.hmdb.ca/proteins/HMDBP00278|website=Human Metabolome database|accessdate=17 February 2015}}</ref>
* [[L-DOPA]] to [[dopamine]] - a [[neurotransmitter]]
* [[5-HTP]] to [[serotonin]] (5-HT) - a neurotransmitter
* [[L-histidine]] to [[histamine]] - a neurotransmitter
* [[phenylalanine]] to [[phenethylamine]] - a [[trace amine]] [[neuromodulator]]
* [[L-tyrosine]] to [[tyramine]] - a trace amine neuromodulator
* [[tryptophan]] to [[tryptamine]] - a trace amine neuromodulator
The enzyme uses [[pyridoxal phosphate]], the active form of [[vitamin B6|vitamin B<sub>6</sub>]], as a [[cofactor (biochemistry)|cofactor]].
{{Phenylalanine biosynthesis|align=left}}

[[File:Serotonin biosynthesis.svg|thumb|right|150px|Human serotonin biosynthesis pathway]]
{{clear left}}

*Below is the DNA sequence that codes for the protein above

 atgttcagcagagagttcagaagaagagggaaagagatggtcttctatatcgccttctat
 ctgttcgggatcgaggggagacccgtctatcccttcgtcgagcccgggtatctgagaccc
 ctgatccccgccacagccccccaagagcccgagacatatgagttcatcatcaaattcatc
 gagaaaatcatcatgcccggggtcacacattggcatagcccctatttcttcgcctatttc
 cccacagccagcagctatcccgccatgctggccttcatgctgtgcggggccatcgggtgc
 atcgggttcagctgggccgccagccccgcctgcacagagctggagacagtcatgatgttc
 tggctggggaaaatgctggagctgcccgaggccttcctggccgggagagccggggagggg
 gggggggtcatccaagggagcgccagcgaggccacactggtcgccctgctggccgccaga
 acaaaagtcatcagacaactgcaagccgccagccccgagttcacacaagccgccatcatg
 gagaaactggtcgcctatacaagcttccaagcccatagcagcgtcgagagagccgggctg
 atcggggggatcaaactgaaagccgtccccagcttcgggaatttcagcatgagagccagc
 gccctgagagaggccctggagagattcaaagccgccgggctgatccccttcttcgtcgtc
 gccacactggggacaacaagctgctgcagcttcttcaatctgctggaggtcgggcccatc
 tgcaatcaagagggggtctggctgcatatcttcgccgcctatgccgggagcgccttcatc
 tgccccgagttcagatatctgctgaatggggtcgagttcgccttcagcttcaatttcaat
 ccccataaatggctgctggtcaatttcttctgcagcgccatgtgggtcaaaagaagaaca
 ttcctgacaggggccttcaatatgttccccgtctatctgaaacatagccatcaattcagc
 gggttcatcacattctatagacattggcaaatccccctggggagaagattcagaagcctg
 aaaatgtggttcgtcttcagaatgtatggggtcaaagggctgcaagcctatatcagaaaa
 catgtcgagctgagccatgagttcgagagcctggtcagacaattccccagattcgagatc
 tgcacagaggtcatcctggggctggtctgcttcagactgaaagggagcaatgagctgaat
 gagacactgctgcaaagaatcaatagcgccaaaaaaatccatctggtcccctgcagactg
 agattcaaattcgtcctgagattcgccgtctgcgccagaacagtcgagagcgcccatgtc
 caactggcctgggagcatatcagcttcctggccagcagcgtcctgagagccgagaaagag

===Construct a digital plasmid===

*Genome and metabolic pathways of E. coli (IMPORTANT!): http://ecocyc.org/ and http://biocyc.org/

[[File:PBR322.svg|thumb|right|150px|plasmid]]
The basic idea is to extend the metabolic pathway of e coli by inserting new genes into it.
The genes code for proteins that make it happen (exactly like described in https://en.wikipedia.org/wiki/N,N-Dimethyltryptamine#Biosynthesis). It is possible to buy synthetic genes on plasmids from the internet for a few hundred dollars. Plasmids are circular strings of DNA that, when bought from synthesizers also often encode for resistance against some form of antibiotics, such as amplifilin. The idea is that after you have inserted the plasmid into a colony of E. coli, you add the antibiotic specific to the plasmid in order to kill off all individual organisms that did not absorb the plasmid. Insertion of genes with the help of a plasmid vector is very well described and can be done in a kitchen. There are youtube tutorials. For example, https://www.youtube.com/watch?v=slY4qrnZIM8.
 
L-Tryptophan, one of the 22 essential amino-acids (which exist in all living bodies), is transformed into DMT through interaction with two enzymes: AAAD (Aromatic L-amino acid decarboxylase) and INMT (Amine N-methyltransferase).
 
E. coli lacks both these enzymes.
 
These enzymes consume co-enzymes that are available in the e.coli.
 
(You can look up the available chemicals in e coli and their relationship with its genome by using ecocyc.org or biocyc.org.)
 
If E.coli contained AAAD and INMT, they would interact with other chemicals present in the cell and produce new chemicals. Going through the list of all chemicals that can interact with the two enzymes, I found that AAAD would produce the phenetylamine and tyramine as a by-product. This is important, as ingesting tyramine in combination with a mono-amine oxidase inhibitor (MAOI) could cause hypertensive crisis. This could be lethal to a psychedelic user ingesting the product.


I took the code for the enzymes from uniprot.org and compiled them into the DNA. For procaryotes this means that one simply translated each amino-acid into codons (DNA-basepair triplets) that code for them.
 
The other parts it took from the biobricks project: An annual competition held by universities to teach students to genetically modify organisms into doing various mostly useful things. (http://parts.igem.org/Catalog?title=Catalog) The biobricks project is also quite a useful place to start off ones own studies, its very pedagogical.
 
The central dogma of genetics is that DNA is transcribed into RNA, that is then translated into sequences of amino-acids. These sequences are proteins, and enzymes are proteins.
 
====BBa_I14033 ====
http://parts.igem.org/Part:BBa_I14033

http://beta.labgeni.us/registries/parts_registry/?part=BBa_I14033

promoter

>BBa_I14033 Part-only sequence. Constitutive Promoter, Medium Transcription (38 bp)
 ggcacgtaagaggttccaactttcaccataatgaaaca

The promoter recruits transcriptional machinery, that attach to the DNA and transcribes it into mRNA.

The transcription strength is perhaps one of the most important knobs one could experiment with. Too high transcription would result in too much of the organisms metabolic system being kidnapped for the manufacture of enzymes, which would slow growth. A too low transcription would result in an organism that does not produce enough amounts of DMT.

====BBa_B0029====
http://parts.igem.org/Part:BBa_B0029

Ribosome Binding Site (RBS)

>BBa_B0029 Part-only sequence (15 bp)
 ttcacacaggaaacc

 
The RBS binds the mRNA to ribosomes, that translates the mRNA into amino acids.
 
The rest of the gene below, with the exception of the terminator, is code that will be translated into enzymes. Both enzymes start with methodine, which is also a start-codon. This means that no "cutting and pasting" is needed. Between the code that translates to the enzymes, there are stop codons and a spacer that does not initiate translation.

*********[[Amine N-methyltransferase]] 

====spacer 1====
Makes the ribosome stop translation. The ribosome continues its travel down the single-helix RNA though, so it will encounter the next enzyme-encoding sequence as well.

> spacer
 actgtattccta

Does nothing at all, just to separate the proteins in space, and give the ribosome a chance to properly emit the INMT before beginning on the AAAD.

*******[[Aromatic L-amino acid decarboxylase]]

 Stop codon tagtag

====BBa_B0012 ====
Terminates translation and makes the ribosome emit the protein.
 
>BBa_B0012 Part-only sequence (41 bp)
 tcacactggctcaccttcgggtgggcctttctgcgtttata


Terminator that forms a physical loop out of the single-helix RNA equivalent. The loop is formed by the palindrome contained within the sequence. Read more about it at biobricks wiki under the topic terminators.

====Final====
The whole sequence looks like this:

<code>
ggcacgtaagaggttccaactttcaccataatgaaacattcacacagga
aaccatggaggggaaagtctatatcgggggggagttctatgagaaagag
ttcacacccaaattctatctgacaacatattatagcttccatagcgggc
ccgtcgccgagcaagagatcgtcaaattcagcctgcaaaatctgtatca
aacattcagcacagggggggtcggggggttcgtcctgatcttcatcggg
agcgggcccacaatctatcaactgctgagcgcctgcgaggtcttcagag
agatcatcgtcacattctatacaccccaaaatctgcaagagctgcaaaa
atggctgaaaaaagagcccggggcctatttctggagcagcatcgtccaa
catgcctgcgagctggaggggttcagaagcagatggcaagagaaagagg
ccaaactgagaagaacagtcacaagagtcctgagatgcttcgtcacaaa
aacaccccccctggggagcgcccaagtccccctggccttctgcgtcctg
acattcctggccatggagtgcgcctgccccttcatcttcacatatagag
ccgccctgagaagactggccgggctgctgaaacccggggggcatctggt
cacactggtcacactgagattccaacattatatggtcgggcccaaaaaa
ttcagcggggtctatctggagaaagaggtcgtcgagaaagccatccaat
tcgccgggtgccaagtcctgaaatgcaattgcgtcagcctgagctatag
cgaggcctattgcagccatttcgggctgtgcttcgtcgtcgccagaaaa
gggcccagctagtagactgtattcctaatgttcagcagagagttcagaa
gaagagggaaagagatggtcttctatatcgccttctatctgttcgggat
cgaggggagacccgtctatcccttcgtcgagcccgggtatctgagaccc
ctgatccccgccacagccccccaagagcccgagacatatgagttcatca
tcaaattcatcgagaaaatcatcatgcccggggtcacacattggcatag
cccctatttcttcgcctatttccccacagccagcagctatcccgccatg
ctggccttcatgctgtgcggggccatcgggtgcatcgggttcagctggg
ccgccagccccgcctgcacagagctggagacagtcatgatgttctggct
ggggaaaatgctggagctgcccgaggccttcctggccgggagagccggg
gagggggggggggtcatccaagggagcgccagcgaggccacactggtcg
ccctgctggccgccagaacaaaagtcatcagacaactgcaagccgccag
ccccgagttcacacaagccgccatcatggagaaactggtcgcctataca
agcttccaagcccatagcagcgtcgagagagccgggctgatcgggggga
tcaaactgaaagccgtccccagcttcgggaatttcagcatgagagccag
cgccctgagagaggccctggagagattcaaagccgccgggctgatcccc
ttcttcgtcgtcgccacactggggacaacaagctgctgcagcttcttca
atctgctggaggtcgggcccatctgcaatcaagagggggtctggctgca
tatcttcgccgcctatgccgggagcgccttcatctgccccgagttcaga
tatctgctgaatggggtcgagttcgccttcagcttcaatttcaatcccc
ataaatggctgctggtcaatttcttctgcagcgccatgtgggtcaaaag
aagaacattcctgacaggggccttcaatatgttccccgtctatctgaaa
catagccatcaattcagcgggttcatcacattctatagacattggcaaa
tccccctggggagaagattcagaagcctgaaaatgtggttcgtcttcag
aatgtatggggtcaaagggctgcaagcctatatcagaaaacatgtcgag
ctgagccatgagttcgagagcctggtcagacaattccccagattcgaga
tctgcacagaggtcatcctggggctggtctgcttcagactgaaagggag
caatgagctgaatgagacactgctgcaaagaatcaatagcgccaaaaaa
atccatctggtcccctgcagactgagattcaaattcgtcctgagattcg
ccgtctgcgccagaacagtcgagagcgcccatgtccaactggcctggga
gcatatcagcttcctggccagcagcgtcctgagagccgagaaagagtag
tagtcacactggctcaccttcgggtgggcctttctgcgtttata
</code>

==Appling the theory to actually modify a organism==
===Buy the gene  on a plasmid from some online shop===
with the knowledge of the genes needed to produce DMT you have to send it to a Online shops selling synthetic DNA and buy it on a plasmid.


Online shops selling synthetic DNA:

* http://www.plasmid.com/
* http://www.genscript.com/gene_synthesis.html
* http://www.genewiz.com/public/gene-synthesis.aspx
*https://www.youtube.com/watch?v=lNttxYdGHs4

*Plasmids and Recombinant DNA Technology https://www.youtube.com/watch?v=2YxTcBimxlw

===Introduce the plasmid into a population of chosen organisms===
with the plasmid in hands you have to introduce it on a organism like E.coli
===Grow organisms assimilate the plasmid===

kill off organisms that did not assimilate the plasmid using a pencilin

===Dry the biomass and harvest it using straight to base, or similiar===
using some of Tek Extractions

==More information==
===Foruns discussions===
There has been a lot of reseach on online forums on how to create a organism that produces dmt using genetic engineering. Here is a list of most important threads on this subject.
 
====DMT-Nexus====
* DMT-Nexus - 7/13/2008 what is the deal with genetic engineering?     https://www.dmt-nexus.me/forum/default.aspx?g=posts&t=1758
* DMT-Nexus - 12/10/2008 DIY Genetic Engineering & DMT production https://www.dmt-nexus.me/forum/default.aspx?g=posts&m=63238
* DMT-Nexus - 6/3/2008  a note on genetic engineering  https://www.dmt-nexus.me/forum/default.aspx?g=posts&m=12907
* DMT-Nexus - 12/10/2008 - DMT and genetic engeneering  https://www.dmt-nexus.me/forum/default.aspx?g=posts&t=2892
* DMT-Nexus - 5/18/2011 - Genetically engineered DMT : https://www.dmt-nexus.me/forum/default.aspx?g=posts&t=21752
* DMT-Nexus - 8/10/2013 - Biosynthesis of DMT by yeast?  https://www.dmt-nexus.me/forum/default.aspx?g=posts&t=47315
* DMT-Nexus - 7/22/2015 - Biosynthesis of DMT with modified e.coli : https://www.dmt-nexus.me/forum/default.aspx?g=posts&t=66341
* DMT-Nexus - 12/14/2015 - GMO/Synthehuasca: Would You Take It?  https://www.dmt-nexus.me/forum/default.aspx?g=posts&t=68577
* DMT-Nexus - 4/5/2016 - Gene Modification for entheogen sustainability. https://www.dmt-nexus.me/forum/default.aspx?g=posts&t=70324
* DMT-Nexus - 5/19/2011 - Biosynthesis of Novel Tryptamines Aspirations. https://www.dmt-nexus.me/forum/default.aspx?g=posts&t=21776

====reddit====
*reddit 2014 - Genetic Engineering and DMT production  https://www.reddit.com/r/genetics/comments/1dhyk8/genetic_engineering_and_dmt_production/
*reddit 2016 - Biosynthesis of DMT with modified e.coli https://www.reddit.com/r/DMT/comments/3e36ck/biosynthesis_of_dmt_with_modified_ecoli/

====shroomery====
*shroomery - 05/26/05 - HOW TO MAKE ANY ORGANISM MAKE DMT/PSILOS https://www.shroomery.org/forums/showflat.php/Number/4227329#4227329
*shroomery - 12/27/12 - Genetically engineering psychedelic plants? https://www.shroomery.org/forums/showflat.php/Number/17455243#17455243
*shroomery - 02/21/06 - Psychedelics produced from GM yeasts - what's your opinion? https://www.shroomery.org/forums/showflat.php/Number/5323478

===Related projects===
*Harvard scientists to make LSD factory from microbes : https://www.theguardian.com/science/blog/2011/jun/21/scientists-make-lsd-from-microbes
*Production of THC by genetically modified bacteria : http://www.cannabis-med.org/english/bulletin/ww_en_db_cannabis_artikel.php?id=329#1
*Nature.com Engineered yeast paves way for home-brew heroin : http://www.nature.com/news/engineered-yeast-paves-way-for-home-brew-heroin-1.17566

[[Category:Science]]