Option 6 情報請求 (Material documentation, GLP …...4 情報提供書類の設定 Material Documentationの内容：配列の設定、クローニング方法、オリゴデザイン、試薬と酵素（メーカー、カタログ番号）、GeneArtの担

1

The world leader in serving science

GeneArtTM人工遺伝子合成ご注文マニュアル

2019年2月版

Option 6_情報請求(Material documentation, GLP-source documentation)

2

目次

◼情報提供書類の追加・・・・・・・・・・・・・・・・・・・・・・・・・・・・・・・・・・・・・・・・・P3

◼情報提供書類の設定・・・・・・・・・・・・・・・・・・・・・・・・・・・・・・・・・・・・・・・・・P4

◼情報提供書類の注意事項・・・・・・・・・・・・・・・・・・・・・・・・・・・・・・・・・・・・・P5

◼情報提供書類の例・・・・・・・・・・・・・・・・・・・・・・・・・・・・・・・・・・・・・・・・・・・P6

◼Material Documentation gene synthesis：人工遺伝子合成のMaterial Documentation ・・・・・・・・・・・・・・・・・・・・・・・P7

◼Material Documentation gene delivery：サブクローニングのMaterial Documentation ・・・・・・・・・・・・・・・・・・・・・・・P13

◼GLP-source Documentation gene synthesis：人工遺伝子合成のGLP-source Documentation ・・・・・・・・・・・・・・・・・・・・・P17

◼GLP-source Documentation gene delivery：サブクローニングのGLP-source Documentation ・・・・・・・・・・・・・・・・・・・・P26

3

情報提供書類の追加

▪ 情報提供書類をご希望の場合、「Additional Services」タブから「Extended Documentation」アイコ

ンを、書類を希望するアイコンの上にドラッグ＆ドロップします。

▪ クリックして設定を開始して下さい。

4

情報提供書類の設定

Material Documentationの内容：配列の設定、クローニング方法、オリゴデザイン、試薬と酵素（メーカー、カタログ番号）、GeneArtの担当者、配送業者、トラッキングNo.

GLP-source Documentationの内容：Material Documentationに作業リスト/SOPリスト、 GeneArtの担当者の文書を追加

弊社で10年間、書類の保存も希望する場合、「Data Storage for,,,,」を選択して下さい。

▪ ご希望の情報提供書類を選択し、「Save」をクリックして下さい。

5

情報提供書類の注意事項

▪ 各種情報提供書類は人工遺伝子合成等、該当試験のデータ等を収集する必要があるため、試験開始と同時に申し込む必要があります。試験開始後に別途注文することはできません。

6

情報提供書類の例

▪ 次ページ以降に書類の例を４つ掲載してあります。

Material Documentation gene synthesis：

人工遺伝子合成のMaterial Documentation

Material Documentation gene delivery：

サブクローニングのMaterial Documentation

GLP-source Documentation gene synthesis：

人工遺伝子合成のGLP-source Documentation

GLP-source Documentation gene delivery：

サブクローニングのGLP-source Documentation

7

Material Documentation gene synthesis：

人工遺伝子合成のMaterial Documentation

———————————————————————————————————— M a t e r i a l D o c u m e n t a t i o n g e n e s y n t h e s i s————————————————————————————————————

GA_number

27.12.2007

30.01.2008

name Dr. Marcus Graf

BU Genesynthesis

address Josef-Engert-Str. 11

D-93053 Regensburg

fon (0)+49-941 942 7618

fax (0)+49-941 942 7608

name analyst_name

source sequence_source

optimized for traget_organism

date 17.12.2007

name ordering_person_name

date 27.12.2007

number no_subfragments

name

name releasing_person_name

date 17.01.2008

provider shipping_provider_name

tracking number #########

date 16.04.2008

Gene analysed by

gene_name

Order number:

Gene name

Customer

Date (start)

subframents_name

customer_name

subfragments

Quality assurance doc by

Estimated delivery date

Production manager

Oligo and subfragment design

shipping

CONFIDENTIAL Page 1 of 5 created: 29.04.2008


2. Production of gene_name

No. Name Used for provider

1 synthesis

2 synthesis

3 synthesis

4 synthesis

5 synthesis

6 synthesis

7 synthesis

8 synthesis

9 synthesis

10 synthesis

11 post processing

12 post processing

13 post processing

14 post processing

Oligonucleotides were produced on a Cerbarus Oligo Synthesizer (Geneart AG) using following chemicals:

nucleotides of each respective subfragment subframents_name on separate menu plates. The oligonucleotides of the different subfragments were pooled seperately (subfragment A, B, etc. ) and processed further on.

Oligonucleotides were produced and post-processed on dates summarized in table 1. A Genesis RSP100 (TECAN, Switzerland) was used to adjust oligonucleotide concentration (measured with GENios, TECAN ) to 25µM (diluted in Tris-buffer) and to pool all oligo-

2.1 Oligonucleotide Production

1. Project design

Oligonucleotide and subfragment design was performed using GeneOptimizerTM software (GENEART, Regensburg, Germany) by ordering_person_name. Briefly, the gene_name gene was divided into no_subfragments almost equally sized subfragments subframents_name. The design took into special accout to elaborate a distict cloning strategy to recreate the full length gene by the fusion of the subfragments.

1.1 Gene design

1.2 Cloning strategy and oligo design

The in silico gene expression analysis and multi-parameter gene optimization of gene_name was performed using GeneOptimizerTM software (GENEART, Regensburg, Germany) by analyst_name.

catalogue #



post- menurun-ID Oligo name synthesis processing plates delivered103808 GA-no Ac1-258 2.1.2008 3.1.2008 3.1.2008 4.1.2008103807 GA-no AL1 2.1.2008 3.1.2008 3.1.2008 4.1.2008103808 GA-no AL10 2.1.2008 3.1.2008 3.1.2008 4.1.2008103808 GA-no AL11 2.1.2008 3.1.2008 3.1.2008 4.1.2008103808 GA-no AL12 2.1.2008 3.1.2008 3.1.2008 4.1.2008103808 GA-no AL13 2.1.2008 3.1.2008 3.1.2008 4.1.2008103808 GA-no AL14 2.1.2008 3.1.2008 3.1.2008 4.1.2008103808 GA-no AL15 2.1.2008 3.1.2008 3.1.2008 4.1.2008103807 GA-no AL2 2.1.2008 3.1.2008 3.1.2008 4.1.2008103807 GA-no AL3 2.1.2008 3.1.2008 3.1.2008 4.1.2008103807 GA-no AL4 2.1.2008 3.1.2008 3.1.2008 4.1.2008103807 GA-no AL5 2.1.2008 3.1.2008 3.1.2008 4.1.2008103807 GA-no AL6 2.1.2008 3.1.2008 3.1.2008 4.1.2008103807 GA-no AL7 2.1.2008 3.1.2008 3.1.2008 4.1.2008103807 GA-no AL8 2.1.2008 3.1.2008 3.1.2008 4.1.2008103808 GA-no AL9 2.1.2008 3.1.2008 3.1.2008 4.1.2008103807 GA-no Am1 2.1.2008 3.1.2008 3.1.2008 4.1.2008103807 GA-no Am10 2.1.2008 3.1.2008 3.1.2008 4.1.2008103807 GA-no Am11 2.1.2008 3.1.2008 3.1.2008 4.1.2008103807 GA-no Am12 2.1.2008 3.1.2008 3.1.2008 4.1.2008103807 GA-no Am13 2.1.2008 3.1.2008 3.1.2008 4.1.2008103807 GA-no Am14 2.1.2008 3.1.2008 3.1.2008 4.1.2008103807 GA-no Am2 2.1.2008 3.1.2008 3.1.2008 4.1.2008103807 GA-no Am3 2.1.2008 3.1.2008 3.1.2008 4.1.2008103807 GA-no Am4 2.1.2008 3.1.2008 3.1.2008 4.1.2008103807 GA-no Am5 2.1.2008 3.1.2008 3.1.2008 4.1.2008103807 GA-no Am6 2.1.2008 3.1.2008 3.1.2008 4.1.2008103807 GA-no Am7 2.1.2008 3.1.2008 3.1.2008 4.1.2008103807 GA-no Am8 2.1.2008 3.1.2008 3.1.2008 4.1.2008103807 GA-no Am9 2.1.2008 3.1.2008 3.1.2008 4.1.2008103808 GA-no Apb 2.1.2008 3.1.2008 3.1.2008 4.1.2008103808 GA-no Apf 2.1.2008 3.1.2008 3.1.2008 4.1.2008103808 GA-no As1-569 2.1.2008 3.1.2008 3.1.2008 4.1.2008103808 GA-no Bc1-974 2.1.2008 3.1.2008 3.1.2008 4.1.2008103808 GA-no Bc2-1058 2.1.2008 3.1.2008 3.1.2008 4.1.2008103808 GA-no BL1 2.1.2008 3.1.2008 3.1.2008 4.1.2008103808 GA-no BL10 2.1.2008 3.1.2008 3.1.2008 4.1.2008103808 GA-no BL11 2.1.2008 3.1.2008 3.1.2008 4.1.2008103808 GA-no BL12 2.1.2008 3.1.2008 3.1.2008 4.1.2008103808 GA-no BL13 2.1.2008 3.1.2008 3.1.2008 4.1.2008103808 GA-no BL14 2.1.2008 3.1.2008 3.1.2008 4.1.2008103808 GA-no BL15 2.1.2008 3.1.2008 3.1.2008 4.1.2008103808 GA-no BL2 2.1.2008 3.1.2008 3.1.2008 4.1.2008103808 GA-no BL3 2.1.2008 3.1.2008 3.1.2008 4.1.2008103808 GA-no BL4 2.1.2008 3.1.2008 3.1.2008 4.1.2008

Table 1: Oligo production and assembly



103808 GA-no BL5 2.1.2008 3.1.2008 3.1.2008 4.1.2008103808 GA-no BL6 2.1.2008 3.1.2008 3.1.2008 4.1.2008103808 GA-no BL7 2.1.2008 3.1.2008 3.1.2008 4.1.2008103808 GA-no BL8 2.1.2008 3.1.2008 3.1.2008 4.1.2008103808 GA-no BL9 2.1.2008 3.1.2008 3.1.2008 4.1.2008103808 GA-no Bm1 2.1.2008 3.1.2008 3.1.2008 4.1.2008103808 GA-no Bm10 2.1.2008 3.1.2008 3.1.2008 4.1.2008103808 GA-no Bm11 2.1.2008 3.1.2008 3.1.2008 4.1.2008103808 GA-no Bm12 2.1.2008 3.1.2008 3.1.2008 4.1.2008103808 GA-no Bm13 2.1.2008 3.1.2008 3.1.2008 4.1.2008103808 GA-no Bm14 2.1.2008 3.1.2008 3.1.2008 4.1.2008103808 GA-no Bm2 2.1.2008 3.1.2008 3.1.2008 4.1.2008103808 GA-no Bm3 2.1.2008 3.1.2008 3.1.2008 4.1.2008103808 GA-no Bm4 2.1.2008 3.1.2008 3.1.2008 4.1.2008103808 GA-no Bm5 2.1.2008 3.1.2008 3.1.2008 4.1.2008103808 GA-no Bm6 2.1.2008 3.1.2008 3.1.2008 4.1.2008103808 GA-no Bm7 2.1.2008 3.1.2008 3.1.2008 4.1.2008103808 GA-no Bm8 2.1.2008 3.1.2008 3.1.2008 4.1.2008103808 GA-no Bm9 2.1.2008 3.1.2008 3.1.2008 4.1.2008103808 GA-no Bpb 2.1.2008 3.1.2008 3.1.2008 4.1.2008103808 GA-no Bpf 2.1.2008 3.1.2008 3.1.2008 4.1.2008103808 GA-no Bs1-1038 2.1.2008 3.1.2008 3.1.2008 4.1.2008103519 GA-no CL1 27.12.2007 28.12.2007 28.12.2007 29.12.2007103519 GA-no CL2 27.12.2007 28.12.2007 28.12.2007 29.12.2007103519 GA-no CL3 27.12.2007 28.12.2007 28.12.2007 29.12.2007103519 GA-no CL4 27.12.2007 28.12.2007 28.12.2007 29.12.2007103519 GA-no CL5 27.12.2007 28.12.2007 28.12.2007 29.12.2007103519 GA-no CL6 27.12.2007 28.12.2007 28.12.2007 29.12.2007103519 GA-no CL7 27.12.2007 28.12.2007 28.12.2007 29.12.2007103519 GA-no Cm1 27.12.2007 28.12.2007 28.12.2007 29.12.2007103519 GA-no Cm2 27.12.2007 28.12.2007 28.12.2007 29.12.2007103519 GA-no Cm3 27.12.2007 28.12.2007 28.12.2007 29.12.2007103519 GA-no Cm4 27.12.2007 28.12.2007 28.12.2007 29.12.2007103519 GA-no Cm5 27.12.2007 28.12.2007 28.12.2007 29.12.2007103519 GA-no Cm6 27.12.2007 28.12.2007 28.12.2007 29.12.2007103519 GA-no Cm7 27.12.2007 28.12.2007 28.12.2007 29.12.2007



Table 2: enzymes, reagents and kits

No. Name Used for provider1 Truescript polymerase

2 RE1 restriction enzyme





3 PNK Polynucleotide kinase

4 Pfu Ligase Ligase

5 T4 DNA Ligase Ligase

6 XL10-Gold E. coli

7 Taq Mastermix polymerase

8 Pepton made from soybean media surrogate

9 Hefeextrakt media surrogate

10 Glucose media surrogate

11 NaCl sodium chloride

12 Midi-Prep DNA-purification kit

13 Mini-Prep DNA-purification kit

14 Gelpurification DNA-purification kit

15 Gelpurification Vector-DNA-purific.

16 96 PCR purification (cPCR) DNA-purification kit

17 96 PCR purification (SPCR) DNA-purification kit

18 Qiaquick 96-Plate DNA-purification kit

19 Big-dye 3.1 SEQ-Kit sequencing kit

19 X-term SEQ-Kit sequencing kit

20 Sephadex G-50 Fine sequence purification

21 96 SEQ purification sequence purification

No BSA was added to enzyme reactions.

3. Qualitiy control and shipping

Regensburg, 29.04.2008Dr. Thomas Zeller

(Leader QM / QC)

The quality assurance documentation was created using GeneOptimizerTM software (GENEART, Regensburg, Germany) by releasing_person_name. The DNA sample including documentation was shipped as indicated using shipping_provider_name (Tracking number: #########).

catalogue #

2.2 Gene synthesis

The subfragments subframents_name were assembled, from the respective oligonucleotides, cloned and sequenced. 100% correct subfragments subframents_name were fused resulting in the full length gene. All used reagents and materials were summarized in table 2. The work was performed according to GENEART's standard operating procedures (DIN EN ISO 9001:2008 certified).


8

Material Documentation gene delivery：

サブクローニングのMaterial Documentation

———————————————————————————————————— M a t e r i a l D o c u m e n t a t i o n g e n e d e l i v e r y————————————————————————————————————

GA number

15.05.2007

31.05.2007

name Ruth Wittmann

BU GeneDelivery


D-93053 Regensburg

fon (0)+49-941 942 7665

fax (0)+49-941 942 7608

name analyst_name

date 15.05.2007

dependent on precursor_number

5 ' site RE1

3 ' site RE2

target vector customer_vector_name

name Dr. Thomas Zeller

date 01.06.2007



date 25.06.2007

Date (start)

customer_name


gene_name

Order number:

Gene name

Customer


Production manager

Subcloning analysed by

shipping





No. Name Used for provider1 CIP dephosphorylation



3 Agarose AGE

4 Pfu Ligase Ligase


6 DH10B E. coli

7 Taq Mastermix PCR

8 LB-Medium media surrogate



13 Ampicillin antibiotic (selection)

14 Kanamycin antibiotic (selection)

15 Spectinimycin antibiotic (selection)



2.1 Oligonucleotide Production (not applicable)

Oligonucleotide and subfragment design for the designated sequnece was performed using GeneOptimizerTM software (GENEART, Regensburg, Germany). The design of gene_name (precursor_number) took into special accout to elaborate a distict cloning strategy to recreate the full length gene and allowing a subsequent cloning step for directional cloning into the custorer vector customer_vector_name via RE1 and RE2 (GA number).

1.1 Gene design


The in silico gene expression analysis and multi-parameter gene optimization of gene_name (precursor_number) was performed using GeneOptimizerTM software (GENEART, Regensburg, Germany).

1. Project design

2.2 Subcloning and plasmid preparation

The synthetic full length gene gene_name was transfered from GENEART standard vector into customer_vector_name by RE1 / RE2. A clone containing a correct sized plasmid was inocculated for midi scale production. The final construct was verified by sequencing. The sequence congruence within the used cloning sites was 100%.All used reagents and materials were summarized in table 2. The work was performed according to GENEART's standard operating procedures (DIN EN ISO 9001:2008 certified).

catalogue #






20 Sephadex G-50 Fine seq. purification

21 96 SEQ purification seq. purification



Regensburg, 29.04.2008

The quality assurance documentation was created using GeneOptimizerTM software (GENEART, Regensburg, Germany) by Dr. Thomas Zeller. The DNA sample including documentation was shipped as indicated using shipping_provider_name (Tracking number: #########).

Dr. Thomas Zeller(Leader QM / QC)


9

GLP-source Documentation gene synthesis：

人工遺伝子合成のGLP-source Documentation

———————————————————————————————————— G L P S o u r c e D o c u m e n t a t i o n g e n e s y n t h e s i s————————————————————————————————————

GA_number

27.12.2007

30.01.2008

name Dr. Marcus Graf

BU Genesynthesis


D-93053 Regensburg

fon (0)+49-941 942 7618

fax (0)+49-941 942 7608

name analyst_name

source sequence_source

optimized for traget_organism

date 17.12.2007

name ordering_person_name

date 27.12.2007

number no_subfragments

name

name releasing_person_name

date 17.01.2008



date 16.04.2008


subfragments


Order number:

Gene name

Customer

Date (start)

Production manager

Gene analysed by

gene_name

Oligo and subfragment design

shipping

subframents_name

customer_name




No. Name Used for provider catalogue # lot #

1 synthesis

2 synthesis

3 synthesis

4 synthesis

5 synthesis

6 synthesis

7 synthesis

8 synthesis

9 synthesis

10 synthesis

11 post processing

12 post processing

13 post processing

14 post processing

The in silico gene expression analysis and multi-parameter gene optimization of gene_name was performed using GeneOptimizerTM software (GENEART, Regensburg, Germany) by analyst_name.

1. Project design

Oligonucleotide and subfragment design was performed using GeneOptimizerTM software (GENEART, Regensburg, Germany) by ordering_person_name. Briefly, the gene_name gene was divided into no_subfragments almost equally sized subfragments subframents_name. The design took into special accout to elaborate a distict cloning strategy to recreate the full length gene by the fusion of the subfragments.

1.1 Gene design


Oligonucleotides were produced and post-processed on dates summarized in table 1. A Genesis RSP100 (TECAN, Switzerland) was used to adjust oligonucleotide concentration (measured with GENios, TECAN ) to 25µM (diluted in Tris-buffer) and to pool all oligo-

2.1 Oligonucleotide ProductionOligonucleotides were produced on a Cerbarus Oligo Synthesizer (Geneart AG) using following chemicals:

nucleotides of each respective subfragment subframents_name on separate menu plates. The oligonucleotides of the different subfragments were pooled seperately (subfragment A, B, etc. ) and processed further on.



post- menurun-ID Oligo name synthesis processing plates delivered103808 GA-no Ac1-258 2.1.2008 3.1.2008 3.1.2008 4.1.2008103807 GA-no AL1 2.1.2008 3.1.2008 3.1.2008 4.1.2008103808 GA-no AL10 2.1.2008 3.1.2008 3.1.2008 4.1.2008103808 GA-no AL11 2.1.2008 3.1.2008 3.1.2008 4.1.2008103808 GA-no AL12 2.1.2008 3.1.2008 3.1.2008 4.1.2008103808 GA-no AL13 2.1.2008 3.1.2008 3.1.2008 4.1.2008103808 GA-no AL14 2.1.2008 3.1.2008 3.1.2008 4.1.2008103808 GA-no AL15 2.1.2008 3.1.2008 3.1.2008 4.1.2008103807 GA-no AL2 2.1.2008 3.1.2008 3.1.2008 4.1.2008103807 GA-no AL3 2.1.2008 3.1.2008 3.1.2008 4.1.2008103807 GA-no AL4 2.1.2008 3.1.2008 3.1.2008 4.1.2008103807 GA-no AL5 2.1.2008 3.1.2008 3.1.2008 4.1.2008103807 GA-no AL6 2.1.2008 3.1.2008 3.1.2008 4.1.2008103807 GA-no AL7 2.1.2008 3.1.2008 3.1.2008 4.1.2008103807 GA-no AL8 2.1.2008 3.1.2008 3.1.2008 4.1.2008103808 GA-no AL9 2.1.2008 3.1.2008 3.1.2008 4.1.2008103807 GA-no Am1 2.1.2008 3.1.2008 3.1.2008 4.1.2008103807 GA-no Am10 2.1.2008 3.1.2008 3.1.2008 4.1.2008103807 GA-no Am11 2.1.2008 3.1.2008 3.1.2008 4.1.2008103807 GA-no Am12 2.1.2008 3.1.2008 3.1.2008 4.1.2008103807 GA-no Am13 2.1.2008 3.1.2008 3.1.2008 4.1.2008103807 GA-no Am14 2.1.2008 3.1.2008 3.1.2008 4.1.2008103807 GA-no Am2 2.1.2008 3.1.2008 3.1.2008 4.1.2008103807 GA-no Am3 2.1.2008 3.1.2008 3.1.2008 4.1.2008103807 GA-no Am4 2.1.2008 3.1.2008 3.1.2008 4.1.2008103807 GA-no Am5 2.1.2008 3.1.2008 3.1.2008 4.1.2008103807 GA-no Am6 2.1.2008 3.1.2008 3.1.2008 4.1.2008103807 GA-no Am7 2.1.2008 3.1.2008 3.1.2008 4.1.2008103807 GA-no Am8 2.1.2008 3.1.2008 3.1.2008 4.1.2008103807 GA-no Am9 2.1.2008 3.1.2008 3.1.2008 4.1.2008103808 GA-no Apb 2.1.2008 3.1.2008 3.1.2008 4.1.2008103808 GA-no Apf 2.1.2008 3.1.2008 3.1.2008 4.1.2008103808 GA-no As1-569 2.1.2008 3.1.2008 3.1.2008 4.1.2008103808 GA-no Bc1-974 2.1.2008 3.1.2008 3.1.2008 4.1.2008103808 GA-no Bc2-1058 2.1.2008 3.1.2008 3.1.2008 4.1.2008103808 GA-no BL1 2.1.2008 3.1.2008 3.1.2008 4.1.2008103808 GA-no BL10 2.1.2008 3.1.2008 3.1.2008 4.1.2008103808 GA-no BL11 2.1.2008 3.1.2008 3.1.2008 4.1.2008103808 GA-no BL12 2.1.2008 3.1.2008 3.1.2008 4.1.2008103808 GA-no BL13 2.1.2008 3.1.2008 3.1.2008 4.1.2008103808 GA-no BL14 2.1.2008 3.1.2008 3.1.2008 4.1.2008103808 GA-no BL15 2.1.2008 3.1.2008 3.1.2008 4.1.2008103808 GA-no BL2 2.1.2008 3.1.2008 3.1.2008 4.1.2008103808 GA-no BL3 2.1.2008 3.1.2008 3.1.2008 4.1.2008103808 GA-no BL4 2.1.2008 3.1.2008 3.1.2008 4.1.2008

Table 1: Oligo production and assembly



103808 GA-no BL5 2.1.2008 3.1.2008 3.1.2008 4.1.2008103808 GA-no BL6 2.1.2008 3.1.2008 3.1.2008 4.1.2008103808 GA-no BL7 2.1.2008 3.1.2008 3.1.2008 4.1.2008103808 GA-no BL8 2.1.2008 3.1.2008 3.1.2008 4.1.2008103808 GA-no BL9 2.1.2008 3.1.2008 3.1.2008 4.1.2008103808 GA-no Bm1 2.1.2008 3.1.2008 3.1.2008 4.1.2008103808 GA-no Bm10 2.1.2008 3.1.2008 3.1.2008 4.1.2008103808 GA-no Bm11 2.1.2008 3.1.2008 3.1.2008 4.1.2008103808 GA-no Bm12 2.1.2008 3.1.2008 3.1.2008 4.1.2008103808 GA-no Bm13 2.1.2008 3.1.2008 3.1.2008 4.1.2008103808 GA-no Bm14 2.1.2008 3.1.2008 3.1.2008 4.1.2008103808 GA-no Bm2 2.1.2008 3.1.2008 3.1.2008 4.1.2008103808 GA-no Bm3 2.1.2008 3.1.2008 3.1.2008 4.1.2008103808 GA-no Bm4 2.1.2008 3.1.2008 3.1.2008 4.1.2008103808 GA-no Bm5 2.1.2008 3.1.2008 3.1.2008 4.1.2008103808 GA-no Bm6 2.1.2008 3.1.2008 3.1.2008 4.1.2008103808 GA-no Bm7 2.1.2008 3.1.2008 3.1.2008 4.1.2008103808 GA-no Bm8 2.1.2008 3.1.2008 3.1.2008 4.1.2008103808 GA-no Bm9 2.1.2008 3.1.2008 3.1.2008 4.1.2008103808 GA-no Bpb 2.1.2008 3.1.2008 3.1.2008 4.1.2008103808 GA-no Bpf 2.1.2008 3.1.2008 3.1.2008 4.1.2008103808 GA-no Bs1-1038 2.1.2008 3.1.2008 3.1.2008 4.1.2008103519 GA-no CL1 27.12.2007 28.12.2007 28.12.2007 29.12.2007103519 GA-no CL2 27.12.2007 28.12.2007 28.12.2007 29.12.2007103519 GA-no CL3 27.12.2007 28.12.2007 28.12.2007 29.12.2007103519 GA-no CL4 27.12.2007 28.12.2007 28.12.2007 29.12.2007103519 GA-no CL5 27.12.2007 28.12.2007 28.12.2007 29.12.2007103519 GA-no CL6 27.12.2007 28.12.2007 28.12.2007 29.12.2007103519 GA-no CL7 27.12.2007 28.12.2007 28.12.2007 29.12.2007103519 GA-no Cm1 27.12.2007 28.12.2007 28.12.2007 29.12.2007103519 GA-no Cm2 27.12.2007 28.12.2007 28.12.2007 29.12.2007103519 GA-no Cm3 27.12.2007 28.12.2007 28.12.2007 29.12.2007103519 GA-no Cm4 27.12.2007 28.12.2007 28.12.2007 29.12.2007103519 GA-no Cm5 27.12.2007 28.12.2007 28.12.2007 29.12.2007103519 GA-no Cm6 27.12.2007 28.12.2007 28.12.2007 29.12.2007103519 GA-no Cm7 27.12.2007 28.12.2007 28.12.2007 29.12.2007



run-ID Oligo name sequence103808 GA-no Ac1-258 NNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNN

103807 GA-no AL1 NNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNN















103807 GA-no Am1 NNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNN














103808 GA-no Apb NNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNN

103808 GA-no Apf NNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNN

103808 GA-no As1-569 NNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNN

103808 GA-no Bc1-974 NNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNN

103808 GA-no Bc2-1058 NNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNN

103808 GA-no BL1 NNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNN

















103808 GA-no Bm1 NNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNN














103808 GA-no Bpb NNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNN

103808 GA-no Bpf NNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNN

103808 GA-no Bs1-1038 NNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNN

103519 GA-no CL1 NNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNN







103519 GA-no Cm1 NNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNN










No. Name Used for provider catalogue # lot#1 Truescript polymerase






3 PNK Polynucleotide kinase

4 Pfu Ligase Ligase


6 XL10-Gold E. coli

7 Taq Mastermix polymerase

8 Pepton made from soybean media surrogate

9 Hefeextrakt media surrogate

10 Glucose media surrogate

11 NaCl sodium chloride






17 96 PCR purification (SPCR) DNA-purification kit

18 Turbo Filter 96 Plates DNA-purification kit



20 Sephadex G-50 Fine sequence purification

21 96 SEQ purification sequence purification


Table 3a: work list for the production of 0723585 subfragments (A, B, ...)No. short description reagents SOP1 1

2 14 or 17

3 3

4 4

5 1

6 14

AA_GS_B4AA_GS_B5AA_GS_B6AA_GS_B8

2.2 Gene synthesis

The subfragments subframents_name were assembled, from the respective oligonucleotides, cloned and sequenced. 100% correct subfragments subframents_name were fused resulting in the full length gene. All used reagents and materials were summarized in table 2. The work was performed according to GENEART's standard operating procedures (DIN EN ISO 9001:2008 certified) as summarized in table 3a and b.

AA_GS_B78AA_GS_B8



7 2

8 14

9 5, 6

10 7

11 16

12 18

13 19, 20, 21

1415 13

16 2

17 14

18 18

19 19, 20, 21

20

Table 3b: work list for the production of 0723585 full lengh gene

21 5, 6

22 7

23 8, 9, 10, 11

24 12

25 18

26 19, 20, 21

2728


Regensburg, 29.04.2008

AA_GS_B52

The quality assurance documentation was created using GeneOptimizerTM software (GENEART, Regensburg, Germany) by releasing_person_name. The DNA sample including documentation was shipped as indicated using shipping_provider_name (Tracking number: #########).

AA_GS_B31

prepare corret DNA for shipping

sequence analysis of 19

sequencing set up of 26seq. purification 27

inocculate correct clone (size controlled) of 22Midi plasmid preparation of 25

sequencing set up of 15

AA_GS_B51AA_GS_B8AA_GS_B29CS_03/12,CS_04/16

AA_GS_B29AA_GS_B31AA_GS_B52AA_GS_B35/38

AA_GS_B26AA_GS_B27

AA_GS_B10AA_GS_B7AA_GS_B12/15

sequencing and seq. purification 18

AA_QC_06AA_QC_01

AA_GS_B12/15AA_GS_B26AA_GS_B33AA_GS_B36AA_GS_B29

sequence analysis of 28

Dr. Thomas Zeller(Leader QM / QC)

SOP: All standard operating procedures (SOP) are well documented according to ISO9001 specifications. However, details of each specified SOP are proprietary and can not be disclosed.


10

GLP-source Documentation gene delivery：

サブクローニングのGLP-source Documentation

———————————————————————————————————— G L P S o u r c e D o c u m e n t a t i o n g e n e d e l i v e r y————————————————————————————————————

GA number

15.05.2007

31.05.2007

name Ruth Wittmann

BU GeneDelivery


D-93053 Regensburg

fon (0)+49-941 942 7665

fax (0)+49-941 942 7608

name analyst_name

date 15.05.2007

dependent on precursor_number

5 ' site RE1

3 ' site RE2

target vector customer_vector_name

name Dr. Thomas Zeller

date 01.06.2007



date 25.06.2007

shipping


Production manager

Subcloning analysed by

gene_name

Order number:

Gene name

Customer

Date (start)

customer_name






No. Name Used for provider catalogue # lot#1 CIP dephosphorylation



3 Agarose AGE

4 Pfu Ligase Ligase


6 DH10B E. coli

7 Taq Mastermix PCR

8 LB-Medium media surrogate



13 Ampicillin antibiotic (selection)

14 Kanamycin antibiotic (selection)

15 Spectinimycin antibiotic (selection)


2.2 Subcloning and plasmid preparation

The synthetic full length gene gene_name was transfered from GENEART standard vector into customer_vector_name by RE1 / RE2. A clone containing a correct sized plasmid was inocculated for midi scale production. The final construct was verified by sequencing. The sequence congruence within the used cloning sites was 100%.

The in silico gene expression analysis and multi-parameter gene optimization of gene_name (precursor_number) was performed using GeneOptimizerTM software (GENEART, Regensburg, Germany).

1. Project design

Oligonucleotide and subfragment design for the designated sequnece was performed using GeneOptimizerTM software (GENEART, Regensburg, Germany). The design of gene_name (precursor_number) took into special accout to elaborate a distict cloning strategy to recreate the full length gene and allowing a subsequent cloning step for directional cloning into the custorer vector customer_vector_name via RE1 and RE2 (GA number).

1.1 Gene design


2.1 Oligonucleotide Production (not applicable)

All used reagents and materials were summarized in table 2. The work was performed according to GENEART's standard operating procedures (DIN EN ISO 9001:2008 certified) as summarized in table 3.







20 Sephadex G-50 Fine seq. purification

21 96 SEQ purification seq. purification


Table 3: work list for the cloning of 0704854

No. short description reagents SOP1 2

2 3, 16, 17

3 1, 4 or 5, 6, 8

4 3, 7

5 8, 13 or 14 or 15

6 10

7 2

8 18

9 19

10 20, 21

1112


Regensburg, 29.04.2008Dr. Thomas Zeller

(Leader QM / QC)

SOP: All standard operating procedures (SOP) are well documented according to ISO9001 specifications. However, details of each specified SOP are proprietary and can not be disclosed.

inocculate medium with correct cloneplasmid preparation ( midi scale) GD_A25

GD_A06, GD_A37GD_A05, GD_A32

GD_A20GD_A22

GD_A07, GD_A09

prepare corret DNA for shipping

sequencing

The quality assurance documentation was created using GeneOptimizerTM software (GENEART, Regensburg, Germany) by Dr. Thomas Zeller. The DNA sample including documentation was shipped as indicated using shipping_provider_name. (Tracking number: #########)

CS_03, CS_12CS_04, CS_16AA_QC_01AA_QC_06

GD_A37GD_B29


Documents

Option 6 情報請求 (Material documentation, GLP …...4 情報提供書類の設定 Material Documentationの内容： 配列の設定、クローニング方法、オリゴデザイン、試薬と酵素（メーカー、カタログ番号）、GeneArtの担

Option 6 情報請求 (Material documentation, GLP …...4 情報提供書類の設定 Material Documentationの内容：配列の設定、クローニング方法、オリゴデザイン、試薬と酵素（メーカー、カタログ番号）、GeneArtの担