Template:BerkiGEM2007 AustinConstructionFiles

From 2007.igem.org

1128 RBS Libraries:
pBca1101-I716051 (built): Kan cassette RBS pool
pBca1101-I716052(built): Cmr cassette RBS pool

Unmutated basic parts:
9145-I716053(built): alpha HbA in 9145(biobricks 2.0 format)
9145-I716054(built): beta HbA in 9145(biobricks 2.0 format)
9145-I716055(built): AHSP in 9145(biobricks 2.0 format)

Cassette Construction Using 1128 RBS Library:
9145-I716056(built): -kan-RBS library-alpha HbA-
9145-I716057(built): -CmR-RBS library-beta HbA-
9145-I716060(built): beta-alpha cassette (with RBS libraries)

9145-I716065(built): 2X mutant -Kan-rbs-alpha Hba (L29F and V96W)
9145-I716066(built): 2X mutant -Cmr-beta Hba (Asn108Lys and Lys82Asp)
9145-I716067(built): 2X mutants -rbs library-beta-rbs library-alpha- (with RBS libraries)

9145-I716061(built): pTRC-rbs-beta-rbs-alpha
pTrc-I716069(built): 2X mutant -pTRC-rbs-2x beta-rbs-2x alpha L29F/V96W and Asn108Lys/Lys82Asp

HbA Subunit Mutant Variants:
9145-I716058(built): 1X mutant HbA alpha (L29F) AKA: I716053m29
9145-I716059(built): 1X mutant HbA beta (Asn108Lys "Presbyterian") AKA: I716054m108
9145-I716062(built): 2X mutant HbA alpha (L29F/V96W) AKA: I716053m29m96
9145-I716063(built): 2X mutant HbA beta (Asn108Lys/Lys82Asp "Providence/Presbyterian") AKA: I716054m108m82
9145-I716072(built): 1X mutant beta Hba (Lys82Asp "Providence")
9145-I716073(built): unmutated Di-alpha fusion, biobricked
9145-I716068(built): 2X mutant Di-Alpha fusion, biobricked

No Scar Rbs-Part RBS library
1103-I716064: -pCon-(no scar site)-rbs-beta-rbs-alpha

9145-I716070(built): alpha HbA-TT
9145-I716077(built): Unmutated Di-Alpha fusion - TT
9145-I716078(built): 2X mutant alpha - TT
9145-I716079(built): 2X mutant Di-alpha fusion -TT

9145-I716080(built): -rbs-(no scar)-beta
9145-I716081(built): -rbs-(no scar)-1X mutant beta (providence Lys82Asp)
9145--I716082(built): -rbs-(no scar)-2X mutant beta (providence/Presbyterian Lys82Asp/Asn108Lys)

9145-I716071(built): -rbs lib-(NO SCAR)-beta-rbs lib-(no scar)-alpha-TT
9145-I716074(built): -rbs lib-(NO SCAR)-beta 1X mutant, providence (Lys82Asp) - rbs lib-(no scar) - unmutated di-alpha-TT- (Best Yield)
9145-I716075(built): -rbs lib-(NO SCAR)-beta 2X mutant, providence (Lys82Asp) and Presbyterian (Asn108Lys)-rbs lib-(no scar) - unmutated di alpha - TT- (Best all around)
9145-I716076(built): -rbs lib-(NO SCAR)-1X mutant beta, presbyterian (Asn108Lys)-rbs lib-(no scar)-2X mutant alpha (L29F and V96W) - TT (Second best all around)

9145-I716083(built): pT7105-rbs lib-(NO SCAR)-beta-rbs lib-(no scar)-alpha-TT
9145-I716084(built): pT7105-rbs lib-(NO SCAR)-beta 1X mutant, providence (Lys82Asp) - rbs lib-(no scar) - unmutated di-alpha-TT- (Best Yield)
9145-I716085(built): pT7105-rbs lib-(NO SCAR)-beta 2X mutant, providence (Lys82Asp) and Presbyterian (Asn108Lys)-rbs lib-(no scar) - unmutated di alpha - TT- (Best all around)
9145-I716086(built): pT7105-rbs lib-(NO SCAR)-1X mutant beta, presbyterian (Asn108Lys)-rbs lib-(no scar)-2X mutant alpha (L29F and V96W) - TT (Second best all around)

petDUET stuffs beta insertion

9145-I716087(built): (unmodified beta)insertion
9145-I716088(built): (Beta providence) insertion
9145-I716089(built): (Beta providence / Presbyterian) insertion
9145-I716090(built): (Beta presbyterian) insertion

petDUET stuffs alpha and beta insertion

9145-I716091(built): unmodified beta + unmodified alpha: "Unmodified" in petDUET
9145-I716092: Beta providence + unmodified dialpha fusion: "Best Yield" in petDUET
9145-I716093(built): Beta providence / Presbyterian + unmodified dialpha fusion : "Best All Around" in petDUET
9145-I716094(built): Beta presbyterian + 2X mutant alpha L29F/V96W: "Second Best All Around" in petDUET

(Basically Biobricked)Hemoglobin cassette + hemA/hemB:

9145-I716095 (built): "Unmodified" (I716091) + hemA/hemB cassette
9145-I716096 (built): "Best Yield" (I716092) + hemA/hemB cassette
9145-I716097 (built): "Best All Around" (I716093) + hemA/hemB cassette
9145-I716098: "Second Best All Around" (I716094) + hemA/hemB cassette

(Basically Biobricked)Hemoglobin cassette + Cytochrome B5/B5 reductase cassette + hemA/hemB cassette:

9145-I716099(built): "Unmodified" (I716091) + cytochrome b5/b5reductase cassette + hemA/hemB cassette
9145-I716100: "Best Yield" (I716092) + cytochrome b5/b5 reductase cassette + hemA/hemB cassette
9145-I716351(built): "Best All Around" (I716093) + cytochrome b5/b5 reductase cassette + hemA/hemB cassette
9145-I716352: "Second Best All Around" (I716094) + cytochrome b5/b5reductase cassette + hemA/hemB cassette


(Basically Biobricked)Hemoglobin cassette + cytochrome b5/b5 reductase + hemA/hemB + sodC/katG cassette:

9145-I716358: "Unmodified" (I716091) + cytochrome b5/b5reductase cassette + hemA/hemB cassette + sodC/KatG
9145-I716359: "Best Yield" (I716092) + cytochrome b5/b5 reductase cassette + hemA/hemB cassette + sodC/katG
9145-I716360: "Best All Around" (I716093) + cytochrome b5/b5 reductase cassette + hemA/hemB cassette + sodC/KatG
9145-I716361: "Second Best All Around" (I716094) + cytochrome b5/b5reductase cassette + hemA/hemB cassette + sodC/KatG


Detox constructs
9145-I716353: AdhE in petDUET

Biobricked petDUET Hemoglobin Cassettes (XhoI/EcoRI/BglII sites removed)
9145-I716354: "Unmodified" (I716091)
9145-I716355: "Best Yield" (I716092)
9145-I716356: "Best All Around" (I716093)
9145-I716357: "Second Best All Around" (I716094)

Different T7's with Hem Genes
9145-I716366: T7 Clone A with Hem genes A + B
9145-I716367: T7 Clone C with Hem genes A + B

Biobricked petDUET Hemoglobin Cassettes + sodC/katG (Line discontinued)
9145-I716362: Unmodified
9145-I716363: "Best Yield" (Mutant Line Discontinuted)
9145-I716364: Best All Around
9145-I716365: "Second Best All Around" (Mutant Line Discontinuted)

Biobricked petDUET Hemoglobin + SodC/KatG + Cytochrome b5/b5 reductase (Line discontinued)
9145-I716368: Unmodified cassette with antioxidants + cytochromes
9145-I716369: Best all around cassette with antioxidants + cytochromes

Biobricked petDUET Hemoglobin + sodC/katG + Cytochromes + AHSP(Line discontinued)
9145-I716370: Unmodified cassette
9145-I716371: Best all around cassette

Biobricked petDUET Hemoglobin + sodC/katG + Cytochromes + AHSP + metAP(Line discontinued)
9145-I716372: Unmodified cassette
9145-I716373: Best all around cassette

Biobricked petDUET Hemoglobin Cassettes + heme
9145-I716374: Unmodified cassette
9145-I716375: Best all around cassette

Biobricked petDUET Hemoglobin Cassettes + heme + Cytochromes
9145-I716376: Unmodified cassette
9145-I716377: Best all around cassette

Biobricked petDUET Hemoglobin Cassettes + heme + AHSP
9145-I716378: Unmodified cassette
9145-I716379: Best all around cassette

Biobricked petDUET Hemoglobin Cassettes + heme + sodC/katG
9145-I716380: Unmodified cassette
9145-I716381: Best all around cassette

Biobricked petDUET Hemoglobin + heme + cytochromes + sodC/katG
9145-I716383: Unmodified
9145-I716384: Best all around

Biobricked petDUET Hemoglobin + heme + cytochromes + sodC/katG + AHSP
9145-I716385: Unmodified
9145-I716386: Best all around

Biobricked petDUET Hemoglobin + heme + cytochromes + sodC/katG + AHSP + metAP
9145-I716386: Unmodified Final Construct
9145-I716387: Best all around Final Construct

Biobricked petDUET Hemoglobin + heme + sodC/katG + AHSP + metAP
9145-I716388: Unmodified
9145-I716389: Best all around

HemeABCD in pBACb
9145-I716382: pBACb with hemABCD

HemeABCD-TT in 9145
9145-I716390: HemABCD-TT in 9145


Unused Numbers


9145-I716391: