Team:NCTU Formosa/Notebook/test6

9/4

Cultivation of

Pcons+RBS+AseI PCR product-1,2,3,4,7,8 Ar

XhoI+PSB1K3 PCR product-1,2 Kr

PSB1A3 Ar

Cultivation plate test

Ar plate and Kr plate

Digestion of

[PBAN-HindIII] Kr*2 (EP)

[PBAN-NheI]Kr*2 (EP)

[PBAN-KpnI-HF]Kr*2 (EP)

[23101+B0034+PBAN (HindIII)]Ar(ES)

[23101+B0034+PBAN (NheI)]Ar(ES)

PCR of

[J23101+B0034+KpnI-HF] Ar*3

Ligation of

[J23101+B0034+KpnI-HF+PSB1C3]

Electrophoresis of

[PBAN-HindIII] Kr*2 (EP)

[PBAN-NheI]Kr*2 (EP)

[PBAN-KpnI-HF]Kr*2 (EP)

[23101+B0034+PBAN (HindIII)]Ar(ES)

[23101+B0034+PBAN (NheI)]Ar(ES)

[23101+B0034+PBAN (HindIII)]Ar(EP)*2

[23101+B0034+PBAN (NheI)]Ar(EP)

[23101+B0034+PBAN (KpnI-HF)]Ar(EP)*2

[23101+B0034+PBAN (KpnI-HF)]Ar PCR*3

9/5

Electrophoresis of

Well 1 Pcons+RBS+AseI--1 (EP) Ar

Well 2 Pcons+RBS+AseI--2 (EP) Ar

Well 3 Pcons+RBS+AseI--3 (EP) Ar

Well 4 Pcons+RBS+AseI--4 (EP) Ar

Well 5 Pcons+RBS+AseI--7 (EP) Ar （DNA sequencing）

Well 6 Pcons+RBS+AseI--8 (EP) Ar

Well 7 Pcons+RBS+NcoI--2 (EP) Cr （DNA sequencing）

Well 8 Pcons+RBS+Ndel--2 (EP) Cr （DNA sequencing）

Well 9 1 Kb marker

Well 10 Pcons+RBS+Ndel--13 (EP) Cr

Well 11 SacI--3 (XP) Kr

Well 12 SacI--4.1 (XP) Kr

Well 13 SacI--4.2 (XP) Kr

Well 14 XhoI--7 (EP) Kr （DNA sequencing）

Well 15 XhoI--8 (EP) Kr（DNA sequencing）

Well 16 SalI--3 (XP) Kr

Mini-prep of

Pcons+RBS+KpnI-HF-7

Pcons+RBS+KpnI-HF-8

Pcons+RBS+KpnI-HF-9

Digestion of

Pcons+RBS+KpnI-HF-7 (EP)

Pcons+RBS+KpnI-HF-8 (EP)

Pcons+RBS+KpnI-HF-9 (EP)

Cultivation of

Pcons+RBS+KpnI-HF Ar PCR product-7,8,9 Ar

Ligation of

[J23101+B0034+HindIII+B0034+PSB1K3] Kr

[J23101+B0034+HindIII+J61048+PSB1K3] Kr

[J23101+B0034+NheI+B0034+PSB1K3] Kr

[J23101+B0034+NheI+J61048+PSB1K3] Kr

Electrophoresis of digested product of

[Pcons+RBS+KpnI-HF]Ar (EP)*3

9/9

Cultivation of

Pcons+RBS+SacI PCR product-16 Ar

Pcons+RBS+SaII PCR product-17 Ar

RBS+BFP+Ter PCR product-16,17,29,30 Ar

Digestion of

XhoI+PSB1K3 (EP)

9/10

Mini-prep of

RBS+BFP+Ter-16 Ar

RBS+BFP+Ter-17 Ar

RBS+BFP+Ter-29 Ar

RBS+BFP+Ter-30 Ar

Pcons+RBS+Sac-16 Ar

Pcons+RBS+Sal-17 Ar

Digestion of

RBS+BFP+Ter-16 Ar(EP)

RBS+BFP+Ter-17 Ar(EP)

RBS+BFP+Ter-29 Ar(EP)

RBS+BFP+Ter-30 Ar(EP)

Pcons+RBS+SacI-16 Ar(EP)

Pcons+RBS+SalI-17 Ar(EP)

Electrophoresis of

Well1：Marker

Well2：Pcons+RBS+SacI-16 dig(EP)

Well3：Pcons+RBS+SalI-17 dig(EP)

Well4：RBS+BFP+Ter-16 dig(EP)

Well5：RBS+BFP+Ter-17 dig(EP)

Well6：RBS+BFP+Ter-29 dig(EP)

Well7：RBS+BFP+Ter-30 dig(EP)

Well8：XhoI-7 dig(XP) Kr

9/11

Digestion of

Pcons+RBS+PBAN(ES)(8 kind except of XhoI)

RBS+BFP+Ter(XP)

Ligation of

Pcons+RBS+XhoI-7 Ar

Cultivation of

Pcons+RBS+AseI-7 Ar

Pcons+RBS+SacI-16 Ar

Pcons+RBS+SalI-17 Ar

Pcons+RBS+NcoI-2 Ar

Pcons+RBS+NdeI-2 Ar

RBS+BFP+Ter-2 Ar

RBS+BFP+Ter-2 Ar

SalI-3 Kr

Ligation of

Pcons+RBS+XhoI-7 Ar

DNA sequencing of

Pcons+RBS+SaeI Ar 8ul VF2

Pcons+RBS+SalI Ar 8ul VF2

RBS+BFP+Ter-29 Ar 8ul VF2 + 8ul VR

RBS+BFP+Ter-30 Ar 5ul VF2 + 5ul VR

Digestion of

Pcons+RBS+HindIII dig(EP)

Pcons+RBS+NheI dig(EP)

Pcons+RBS+NcoI dig(EP)-->9/11 mini

Pcons+RBS+NdeI dig(EP)

Pcons+RBS+SacI dig(EP)

Pcons+RBS+SalI dig(EP)

Pcons+RBS+AseI dig(EP)

Pcons+RBS+KpnI dig(EP)

Ligation of

Pcons+RBS+BFP+Ter-30 Kr

9/12

Transformation of

Pcons+RBS+NheI+RBS+BFP+Ter Kr

Pcons+RBS+NdeI+RBS+BFP+Ter Kr

Pcons+RBS+HindIII+RBS+BFP+Ter Kr

Pcons+RBS+KpnI-HF+RBS+BFP+Ter Kr

Pcons+RBS+AseI+RBS+BFP+Ter Kr

Pcons+RBS+SacI+RBS+BFP+Ter Kr

Pcons+RBS+NcoI+RBS+BFP+Ter Kr

Pcons+RBS+SalI+RBS+BFP+Ter Kr

Pcons+RBS+BFP+Ter Kr

Pcons+RBS+XhoI Ar

Electrophoresis of

Well1：Marker

Well2：Pcons+RBS+NdeI dig(EP) Cr

Well3：Pcons+RBS+NcoI dig(EP) Cr

Well4：Pcons+RBS+SacI dig(EP) Ar

Well5：Pcons+RBS+SalI dig(EP) Ar

Well6：Pcons+RBS+AseI dig(EP) Ar

Well7：Pcons+RBS+HindIII dig(EP) Ar

Well8：Pcons+RBS+NheI dig(EP) Ar

Well9：Pcons+RBS+KpnI dig(EP) Ar

9/13

Ligation of

Pcons+RBS+AseI Kr

Pcons+RBS+NdeI Kr

Pcons+RBS+SalI Kr

Pcons+RBS+SacI Kr

PCR of

Pcons+RBS+HindIII+RBS+BFP+Ter Kr x8

Pcons+RBS+NheI+RBS+BFP+Ter Kr x8

Pcons+RBS+NdeI+RBS+BFP+Ter Kr x8

Pcons+RBS+NcoI+RBS+BFP+Ter Kr x8

Pcons+RBS+KpnI-HF+RBS+BFP+Ter Kr x8

Pcons+RBS+AseI+RBS+BFP+Ter Kr x8

Pcons+RBS+Xhol Kr x8

Pcons+RBS+SalI+RBS+BFP+Ter Kr x16

Pcons+RBS+SacI+RBS+BFP+Ter Kr x16

Electrophoresis of

Gel 1

Pcons+RBS+SalI+RBS+BFP+Ter Kr x16

Gel 2

Pcons+RBS+SacI+RBS+BFP+Ter Kr x16

Gel 3

Pcons+RBS+HindIII+RBS+BFP+Ter Kr x8

Pcons+RBS+NheI+RBS+BFP+Ter Kr x8

Gel 4

Pcons+RBS+NdeI+RBS+BFP+Ter Kr x8

Pcons+RBS+NcoI+RBS+BFP+Ter Kr x8

Gel 5

Pcons+RBS+KpnI-HF+RBS+BFP+Ter Kr x8

Pcons+RBS+AseI+RBS+BFP+Ter Kr x8

Gel 6

Pcons+RBS+Xhol Kr x8

psb1K3 dig(EP)

9/16

Transformation of

psb3K3 *2

psb3C3

Digestion of

psb3K3 dig(EP)

psb1C3 dig(EP)

Cultivation of

Pcons+RBS+PBAN+RBS+BFP+Ter x8

9/17

Mini-prep of

PBAN：XhoI

SalI

SacI

KpnI

Ndel

AseI

NcoI

NheI

HindIII (fail)

Pcons+RBS+PBAN+RBS+BFP+Ter：AseI-4

Ndel-8

HindIII-4 (fail)

SacI-9(fail)

KpnI-1

NcoI-2

NheI-1

Digestion of

PBAN：XhoI

SalI

SacI

KpnI

Ndel

AseI

NcoI

AseI

Pcons+RBS+PBAN+RBS+BFP+Ter：AseI-4

Ndel-8

KpnI-1

NcoI-2

NheI-1

Electrophoresis of

Pcons+RBS+PBAN+RBS+BFP+Ter (Ndel)

Pcons+RBS+PBAN+RBS+BFP+Ter (NcoI)

Pcons+RBS+PBAN+RBS+BFP+Ter (SacI)

Pcons+RBS+PBAN+RBS+BFP+Ter (HindIII)

Pcons+RBS+PBAN+RBS+BFP+Ter (SalI)

Pcons+RBS+PBAN+RBS+BFP+Ter (NheI)

Pcons+RBS+PBAN+RBS+BFP+Ter (KpnI)

Pcons+RBS+PBAN+RBS+BFP+Ter (AseI)

PBAN AseI

PBAN Ndel

PBAN KpnI

PBAN SacI

PBAN SalI

PBAN XhoI

Pcons+RBS+XhoI-1 dig(EP)

Pcons+RBS+Xhol-1 dig(ES)

psb1C3 dig(ES)

Cultivation of

Pcons+RBS+PBAN+RBS+BFP+Ter (SacI)-4.12

Pcons+RBS+PBAN+RBS+BFP+Ter (HindIII)-6

Pcons+RBS+PBAN+RBS+BFP+Ter (SalI)-9

Digestion of

B0034 Kr dig(ES)

Pcons+RBS+AseI+RBS+BFP+Ter Kr dig(EP)

Pcons+RBS+NdeI+RBS+BFP+Ter Kr dig(EP)

NcoI Kr dig(EP)

Ndel Kr dig(EP)

SalI Kr dig(EP)

9/18

Electrophoresis of

Well1：Marker

Well2：B0034 Kr dig(ES)

Well3：Pcons+RBS+AseI+RBS+BFP+Ter Kr dig(EP)

Well4：Pcons+RBS+NdeI+RBS+BFP+Ter Kr dig(EP)

Well5：NcoI Kr dig(EP)

Well6：Ndel Kr dig(EP)

Well7：SalI Kr dig(EP)

Electrophoresis of

Well1：Marker

Well2：B0034 Kr dig(ES)

Well3：Pcons+RBS+AseI+RBS+BFP+Ter Kr dig(EP)

Well4：Pcons+RBS+NdeI+RBS+BFP+Ter Kr dig(EP)

Well5：NcoI Kr dig(EP)

Well6：Ndel Kr dig(EP)

Well7：SalI Kr dig(EP)

Mini-prep of

Pcons+RBS+HindIII+RBS+BFP+Ter-6

Pcons+RBS+SalI+RBS+BFP+Ter-9

Pcons+RBS+SacI+RBS+BFP+Ter-4

Pcons+RBS+SacI+RBS+BFP+Ter-11

Ligation of

Pcons+RBS+AseI+RBS+BFP+Ter Cr

Pcons+RBS+NdeI+RBS+BFP+Ter Cr

NheI Cr

NcoI Cr

9/20

Transformation of

AseI mini Cr

Electrophoresis of

Well1：Marker

Well2：HindIII-2 Kr dig(EP)

Well3：SacI Kr dig(EP)

Well4：Pcons+RBS+HindIII-6+RBS+BFP+Ter Kr dig(EP)

Well5：Pcons+RBS+SacI-11+RBS+BFP+Ter Kr dig(EP)

Well6：Pcons+RBS+SacI-9+RBS+BFP+Ter Kr dig(EP)

Well7：RBS+BFP+Ter Ar dig(XP)

Well8：Pcons+RBS+NheI+RBS+BFP+Ter Kr dig(EP)

9/21

Electrophoresis of PCR products of

Gel 1

Well1：Marker

Well2：3H1

Well3：3H2

Well4：3H3

Well5：3H4

Well6：3H5

Well7：3H6

Well8：Marker

Well9：6K1

Well10：6K2

Well11：6K3

Well12：6K4

Well13：6K5

Well14：6K6

Well15：3Sl1

Well16：3Sl2

Well17：3Sl3

Gel 2

Well1：Marker

Well2：6Sl4

Well3：6Sl5

Well4：3Nh1

Well5：3Nh2

Well6：3Nh3

Well7：3Nh4

Well8：Marker

Well9：3Nh5

Well10：3Nh6

Well11：6X1

Well12：6X2

Well13：6X3

Well14：6X4

Well15：6X5

Well16：6X6

Well17：1K1

Gel 3

Well1：Marker

Well2：1K2

Well3：1K3

Well4：1K4

Well5：1K5

Well6：1K6

Well7：1X1

Well8：Marker

Well9：1X2

Well10：1X3

Well11：1X4

Well12：1X5

Well13：1X6

Well14：3K1

Well15：3K2

Well16：3K3

Well17：3K4

Gel 4

Well1：Marker

Well2：3K5

Well3：3K6

Well4：3Sc1

Well5：3Sc2

Well6：3Sc3

Well7：3Sc4

Well8：Marker

Well9：3Sc5

Well10：3Sc6

Well11：1Sc1

Well12：1Sc2

Well13：1Sc3

Well14：1Sc4

Well15：1Sc5

Well16：1Sc6

Well17：1Nc1

Gel 5

Well1：Marker

Well2：1Nc2

Well3：1Nc3

Well4：1Nc4

Well5：1Nc5

Well6：1Nc6

Well7：1A1

Well8：Marker

Well9：1A2

Well10：1A3

Well11：1A4

Well12：1A5

Well13：1A6

Well14：1Nd1

Well15：1Nd2

Well16：1Nd3

Well17：1Nd4

Gel 6

Well1：Marker

Well2：1Nd5

Well3：1Nd6

Well4：3X1

Well5：3X2

Well6：3X3

Well7：3X4

Well8：Marker

Well9：3X5

Well10：3X6

Well11：3A1

Well12：3A2

Well13：3A3

Well14：3A4

Well15：3A5

Well16：3A6

Cultivation of

KpnI-1

NdeI-1

XhoI-1

SacI-1

AseI-1

Pcons+RBS+HindIII-1

Pcons+RBS+KpnI-4

Pcons+RBS+NheI-4

Pcons+RBS+SalI-5

Pcons+RBS+AseI-5

Pcons+RBS+XhoI-1

Pcons+RBS+SacI-1

Pcons+RBS+KpnI-1+RBS+BFP+Ter

Pcons+RBS+XhoI-6+RBS+BFP+Ter

Digestion of

SalI Kr dig(EP)

Ligation of

NdeI dig(EP)+C backbone

9/22

Transformation of

NheI Cr

Pcons+RBS+NheI+RBS+BFP+Ter Cr

Pcons+RBS+NcoI+RBS+BFP+Ter Cr

Pcons+RBS+Ndel+RBS+BFP+Ter Cr

Pcons+RBS+AseI+RBS+BFP+Ter Cr

Digestion of

HindIII(EP)

Cultivation of

SacI-3 Kr

NcoI-7 Cr

NcoI-8 Cr

Pcons+RBS+SacI-2

Pcons+RBS+XhoI-5+RBS+BFP+Ter

pbs3C5

Ligation of

Pcons+RBS+NcoI+RBS+BFP+Ter Cr

Pcons+RBS+HindIII+RBS+BFP+Ter Cr

Pcons+RBS+SacI+RBS+BFP+Ter Cr

Pcons+RBS+SalI+RBS+BFP+Ter Cr

9/23

Electrophoresis of

Well1：Marker

Well2：KpnI-1 Cr dig(EP)

Well3：NdeI-1 Cr dig(EP)

Well4：XhoI-1 Cr dig(EP)

Well5：SacI-1 Cr dig(EP)

Well6：AseI-1 Cr dig(EP)

Well7：Pcons+RBS+HindIII-1 dig(EP)

Well8：Pcons+RBS+KpnI-4 dig(EP)

Well9：Pcons+RBS+NheI-4 dig(EP)

Well10：Pcons+RBS+AseI-5 dig(EP)

Well11：Pcons+RBS+XhoI-1 dig(EP)

Well12：Pcons+RBS+SacI-1 dig(EP)

Well13：Pcons+RBS+KpnI-7+RBS+BFP+Ter dig(EP)

Well14：SalI Kr dig(EP) 9/21

Well15：SalI Kr dig(EP) 9/19

DNA sequencing of

KpnI-1 Cr dig(EP)

NdeI-1 Cr dig(EP)

XhoI-1 Cr dig(EP)

SacI-1 Cr dig(EP)

AseI-1 Cr dig(EP)

Pcons+RBS+HindIII-1 dig(EP)

Pcons+RBS+KpnI-4 dig(EP)

Pcons+RBS+NheI-4 dig(EP)(fail)

Pcons+RBS+AseI-5 dig(EP)

Pcons+RBS+XhoI-1 dig(EP)

Pcons+RBS+SacI-1 dig(EP)

Pcons+RBS+KpnI-7+RBS+BFP+Ter dig(EP)

Transformation of

Pcons+RBS+HindIII+RBS+BFP+Ter

Pcons+RBS+NcoI+RBS+BFP+Ter

Pcons+RBS+SalI+RBS+BFP+Ter

Pcons+RBS+SacI+RBS+BFP+Ter

9/24

Cultivation of

Pcons+RBS+NheI+RBS+BFP+Ter Cr

Pcons+RBS+AseI+RBS+BFP+Ter Cr

Pcons+RBS+NcoI+RBS+BFP+Ter Cr

Pcons+RBS+NdeI+RBS+BFP+Ter Cr

NheI Cr

psb3C5 Cr

Cultivation of

Pcons+RBS+NheI+RBS+BFP+Ter Cr

Pcons+RBS+AseI+RBS+BFP+Ter Cr

Pcons+RBS+NcoI+RBS+BFP+Ter Cr

Pcons+RBS+NdeI+RBS+BFP+Ter Cr

NheI Cr

psb3C5 Cr

Electrophoresis of

Well1：Marker

Well2：SalI Kr dig(EP)

Well3：Pcons+RBS+XhoI+RBS+BFP+Ter Cr dig(EP)

Well4：Pcons+RBS+SacI Cr dig(EP)

Well5：NcoI Cr dig(EP)

Digestion of

Pcons+RBS+XhoI+RBS+BFP+Ter Cr dig(EP)

Pcons+RBS+SacI-2 Cr dig(EP)

NcoI-8 Cr dig(EP)

SalI-3 Kr dig(EP)

Ligation of

HindIII+psb1C3

PCR of

SacI x8

SalI x5

HindIII x2

NheI x8

Mini-prep and digestion of

Pcons+RBS+AseI+RBS+BFP+Ter

Pcons+RBS+NheI+RBS+BFP+Ter

Pcons+RBS+NdeI+RBS+BFP+Ter

Pcons+RBS+NcoI+RBS+BFP+Ter

psb3C5

NheI

9/25

Electrophoresis and DNA sequencing of

Well1：Marker

Well2：NheI (fail)

Well3：Pcons+RBS+AseI+RBS+BFP+Ter

Well4：Pcons+RBS+NcoI+RBS+BFP+Ter

Well5：Pcons+RBS+NdeI+RBS+BFP+Ter (fail)

Transformation of

HindIII

SalI

Pcons+RBS+SacI+RBS+BFP+Ter

Pcons+RBS+SalI+RBS+BFP+Ter

Pcons+RBS+HindIII+RBS+BFP+Ter

PCR of

NcoI x5(9.10.11.12.13)

Pcons+RBS+SacI x5(7.8.9.10.11)

Cultivation of

NheI

Pcons+RBS+SacI-2(9/22)

Pcons+RBS+NcoI-8(9/22)

Pcons+RBS+NheI-4

Pcons+RBS+NdeI+RBS+BFP+Ter x2

Pcons+RBS+NheI+RBS+BFP+Ter x2

9/26

Mini-prep and digestion(EP) of

NcoI

NheI

Pcons+RBS+SacI-2

Pcons+RBS+NheI-4

Pcons+RBS+NdeI-1+RBS+BFP+Ter

Pcons+RBS+NdeI-2+RBS+BFP+Ter

Electrophoresis of

Well1：Marker

Well2：Pcons+RBS+NdeI-1+RBS+BFP+Ter

Well3：Pcons+RBS+NdeI-2+RBS+BFP+Ter

Well4：NcoI

Well5：NheI

Well6：Pcons+RBS+NheI-4

Well7：Pcons+RBS+SacI-2

Cultivation of

psb3C5 x2

HindIII-3

HindIII-4

SalI-3

SalI-4

Pcons+RBS+HindIII-3+RBS+BFP+Ter

Pcons+RBS+HindIII-4+RBS+BFP+Ter

Pcons+RBS+SalI-3+RBS+BFP+Ter

Pcons+RBS+SalI-4+RBS+BFP+Ter

Pcons+RBS+SacI-3+RBS+BFP+Ter

Pcons+RBS+SacI-4+RBS+BFP+Ter

Pcons+RBS+NcoI-3+RBS+BFP+Ter

Pcons+RBS+NcoI-4+RBS+BFP+Ter

9/28

Electrophoresis of

Well1：HindIII-3

Well2：HindIII-4

Well3：SalI-3

Well4：SalI-4

Well5：Pcons+RBS+SacI-3+RBS+BFP+Ter

Well6：Marker

Well7：Pcons+RBS+NcoI-3+RBS+BFP+Ter

Well8：Pcons+RBS+NcoI-4+RBS+BFP+Ter

Well9：Pcons+RBS+SalI-3+RBS+BFP+Ter

Well10：Pcons+RBS+SalI-4+RBS+BFP+Ter

Well11：psb3C5

9/29

Electrophoresis of

Well1：Marker

Well2：Pcons+RBS+HindIII+RBS+BFP+Ter

Well3：Pcons+RBS+HindIII+RBS+BFP+Ter

Well4：Pcons+RBS+HindIII+RBS+BFP+Ter

Well5：Pcons+RBS+HindIII+RBS+BFP+Ter

Well6：Pcons+RBS+HindIII+RBS+BFP+Ter

Well7：Pcons+RBS+HindIII+RBS+BFP+Ter

Well8：Pcons+RBS+NcoI+RBS+BFP+Ter

Well9：Pcons+RBS+NcoI+RBS+BFP+Ter

Well10：Pcons+RBS+NcoI+RBS+BFP+Ter

Well11：Pcons+RBS+NcoI+RBS+BFP+Ter

Well12：Pcons+RBS+NcoI+RBS+BFP+Ter

Well13：Pcons+RBS+NcoI+RBS+BFP+Ter

Well14：Pcons+RBS+SalI+RBS+BFP+Ter

Well15：Pcons+RBS+SalI+RBS+BFP+Ter

Well16：Pcons+RBS+SalI+RBS+BFP+Ter

Well17：Pcons+RBS+SalI+RBS+BFP+Ter

Well18：Pcons+RBS+SalI+RBS+BFP+Ter

Well19：Pcons+RBS+SalI+RBS+BFP+Ter

Well20：psb3C5 dig

Well21：Marker

Ligation of

Pcons+RBS+KpnI + pbs3C5

Pcons+RBS+SacI + pbs3C5

Pcons+RBS+AseI + pbs3C5

Pcons+RBS+HindIII + pbs3C5

Pcons+RBS+NheI + pbs3C5

Pcons+RBS+SalI + pbs3C5

Pcons+RBS+NcoI + pbs3C5

Pcons+RBS+NdeI + pbs3C5

Pcons+RBS+XhoI+RBS+BFP+Ter + pbs1C3

Pcons+RBS+NdeI+RBS+BFP+Ter + pbs1C3

Pcons+RBS+SacI + pbs1C3

Pcons+RBS+NcoI + pbs1C3

DNA sequencing of

SalI-4

HindIII-4

Pcons+RBS+SalI-3+RBS+BFP+Ter

Pcons+RBS+SalI-2

Pcons+RBS+NheI-4

NheI(9/26)

Pcons+RBS+HindIII+RBS+BFP+Ter(9/30)

Pcons+RBS+SalI+RBS+BFP+Ter(9/30)

9/30

PCR and Electrophoresis of

Pcons+RBS+NdeI+RBS+BFP+Ter

NcoI

Pcons+RBS+NheI+RBS+BFP+Ter

Pcons+RBS+XhoI+RBS+BFP+Ter

Pcons+RBS+SacI+RBS+BFP+Ter

Cultivation of

Pcons+RBS+NheI-11+RBS+BFP+Ter

Pcons+RBS+NheI-13+RBS+BFP+Ter

Pcons+RBS+NdeI-3+RBS+BFP+Ter

Pcons+RBS+NdeI-10+RBS+BFP+Ter

Pcons+RBS+XhoI-3+RBS+BFP+Ter

Pcons+RBS+XhoI-5+RBS+BFP+Ter

Pcons+RBS+SacI-2+RBS+BFP+Ter

Pcons+RBS+SacI-11+RBS+BFP+Ter

NcoI-3

NcoI-12