Revision 62022-03-22 - IsaacGifford

META TOPICPARENT	name="ProtocolList"

Acinetobacter baylyi ADP1 Overview

ADP1 Resources

Genome Sequences
- ADP1 type strain (GenBank Format)
  In general, use this genome as a reference when referring to coordinates of genes and features in the ADP1 genome. We added the locations of IS1236 elements to the linked sequence. Otherwise it is the same as the one on NCBI.
  - Original sequence of the ADP1 type strain at NCBI (GenBank: NC_005966.1)
  - BLAST a sequence against this genome at NCBI
- ADP1 Barrick Lab strain (GFF3 Format)
  Incorporates a small number of mutations found in the ADP1 strain that the Barrick lab originally obtained from Valérie de Crécy-Lagard. Use for analyzing NGS sequencing results.
- ADP1-ISx (GFF3 Format)
  Strain created in the Barrick lab with all IS1236 elements deleted.

Other Useful Sequences
- pBTK622
  Plasmid containing the tdk-kanR cassette used for genome editing.

Functional Genomics

Changed:

<
<

- ADP1 pathways and gene functions (KEGG)
- ADP1 pathways and gene functions in (BioCyc)

>
>

- ADP1 pathways and gene functions in KEGG
- ADP1 pathways and gene functions in BioCyc

Working with ADP1

Culture conditions

Changed:

<
<

- LB is typically used when growing ADP1 in rich media and Minimal Succinate (ABMS) media is used when growing ADP1 in minimal media (recipes for both medias can be found here).
  - Media can be supplemented with antibiotics for selection of strains transformed with resistance markers as described below.

>
>

- LB is typically used when growing ADP1 in rich media.
- Minimal Succinate (ABMS) media is used when growing ADP1 in minimal media.

Added:

>
>

- Media can be supplemented with antibiotics for selection of strains transformed with resistance markers as described below.

- ADP1 grows at both 30°C and 37°C. The Barrick Lab typically incubates ADP1 cultures at 30°C with shaking at 200 rpm.

Storage
- Cultures of ADP1 can be stored for a few days at 4°C.
- Glycerol or DMSO can be used as cryoprotectants when freezing cultures stocks (using the protocol here)

Genomic DNA extraction: The Barrick Lab uses a PureLink Genomic DNA Mini Kit for genomic DNA extractions. 30 minutes is sufficient incubation time at 55°C for lysing cultures of ADP1.

Protocols: the following protocols are commonly used when working with ADP1
- PCR Amplifying DNA.
- Golden Transformation Creating a construct for integration into the ADP1 genome with Golden Gate ligation and transforming it into ADP1. Constructs can be used for gene deletion, inserting DNA sequences, or introducing mutations.
- Growth curves Calculating growth rates using a plate reader.
- Competence assay Calculating transformation rates and confirming competence of ADP1 mutants.

Antibiotic Resistance

ADP1 is intrinsically resistant to beta-lactam antibiotics. Do not use plasmids/constructs that have beta-lactamase (bla) as the selectable marker gene for ampicillin (or carbenicillin) resistance. Most other common antibiotic selection markers can be used.

We most commonly use kanamycin and spectinomycin resistance markers. The rate of spontaneous mutations to KanR is undetectable. SpecR mutants can arise at a low frequency (~10⁻⁸ to 10⁻⁹) due to chromosomal mutations, so only use this marker for transformations that have a significantly higher frequency than this.

Antibiotic	MIC	Standard Conc.	Status	Notes
Ampicillin (or carbenicillin)	128 µg/ml	100 µg/ml	Resistant
Kanamycin	12.5-6.25 µg/ml (3.125µg/mL doesn't inhibit)	50 µg/ml	Sensitive	MIC info from Barrick Lab results (G.S.)
Spectinomycin	?	60 µg/ml	Sensitive	Spontaneous resistance
Streptomycin	6.25-12.5 µg/ml (escapes observed at 6.25µg/mL)	25 µg/ml	Sensitive	Barrick Lab results (G.S.)
Chloramphenicol	4.0 µg/ml	20 µg/ml	Sensitive	Spontaneous resistance
Gentamycin	1.3 µg/ml	5 µg/ml	Sensitive
Tetracycline	1.0 µg/ml	10 µg/ml	Sensitive
Rifampicin	0.6 µg/ml	100 µg/ml	Sensitive	Spontaneous resistance
Nalidixic acid (mutagen/causes DNA double strand breaks)	1.9 µg/ml (no growth at 24 hr, but growth at 48 hr) (1 µg/ml doesn't inhibit)	3.75 µg/ml	Sensitive	Barrick Lab results (G.S.)

Source: Gomez MJ, Neyfakh AA. (2006) Genes involved in intrinsic antibiotic resistance of Acinetobacter baylyi. Antimicrob Agents Chemother 50:3562–3567. DOI: 10.1128/AAC.00579-06

Plasmids

Common high-copy number E. coli plasmids do not work in ADP1.

A synthetic plasmid (pBAV1K) and broad-host-range plasmid (RSF1010) have been used successfully. In our hands, pBAV1K is less reliable. It is also important to note that because plasmid DNA is linearized when taken up through the competence apparatus, that transformation of plasmids can be significantly less efficient than transformations used to incorporate DNA directly in the chromosome.

Origin	Copy Number	Example
ColE1 (pUC, pBR322, etc.) type plasmids	Do not replicate	NA
pBAV1K	Low-medium copy number	pBAV1K-T5-gfp
RSF1010	Low-medium copy number	pBTK403

Revision 52022-03-22 - JeffreyBarrick

META TOPICPARENT	name="ProtocolList"

Acinetobacter baylyi ADP1 Overview

ADP1 Resources

Genome Sequences
- ADP1 type strain (GenBank Format)
  In general, use this genome as a reference when referring to coordinates of genes and features in the ADP1 genome. We added the locations of IS1236 elements to the linked sequence. Otherwise it is the same as the one on NCBI.
  - Original sequence of the ADP1 type strain at NCBI (GenBank: NC_005966.1)
  - BLAST a sequence against this genome at NCBI
- ADP1 Barrick Lab strain (GFF3 Format)
  Incorporates a small number of mutations found in the ADP1 strain that the Barrick lab originally obtained from Valérie de Crécy-Lagard. Use for analyzing NGS sequencing results.
- ADP1-ISx (GFF3 Format)
  Strain created in the Barrick lab with all IS1236 elements deleted.

Other Useful Sequences
- pBTK622
  Plasmid containing the tdk-kanR cassette used for genome editing.

Functional Genomics
- ADP1 pathways and gene functions (KEGG)
- ADP1 pathways and gene functions in (BioCyc)

Working with ADP1

Culture conditions
- LB is typically used when growing ADP1 in rich media and Minimal Succinate (ABMS) media is used when growing ADP1 in minimal media (recipes for both medias can be found here).
  - Media can be supplemented with antibiotics for selection of strains transformed with resistance markers as described below.
- ADP1 grows at both 30°C and 37°C. The Barrick Lab typically incubates ADP1 cultures at 30°C with shaking at 200 rpm.

Storage
- Cultures of ADP1 can be stored for a few days at 4°C.
- Glycerol or DMSO can be used as cryoprotectants when freezing cultures stocks (using the protocol here)

Genomic DNA extraction: The Barrick Lab uses a PureLink Genomic DNA Mini Kit for genomic DNA extractions. 30 minutes is sufficient incubation time at 55°C for lysing cultures of ADP1.

Protocols: the following protocols are commonly used when working with ADP1
- PCR Amplifying DNA.
- Golden Transformation Creating a construct for integration into the ADP1 genome with Golden Gate ligation and transforming it into ADP1. Constructs can be used for gene deletion, inserting DNA sequences, or introducing mutations.
- Growth curves Calculating growth rates using a plate reader.
- Competence assay Calculating transformation rates and confirming competence of ADP1 mutants.

Antibiotic Resistance

ADP1 is intrinsically resistant to beta-lactam antibiotics. Do not use plasmids/constructs that have beta-lactamase (bla) as the selectable marker gene for ampicillin (or carbenicillin) resistance. Most other common antibiotic selection markers can be used.

We most commonly use kanamycin and spectinomycin resistance markers. The rate of spontaneous mutations to KanR is undetectable. SpecR mutants can arise at a low frequency (~10⁻⁸ to 10⁻⁹) due to chromosomal mutations, so only use this marker for transformations that have a significantly higher frequency than this.

Antibiotic	MIC	Standard Conc.	Status	Notes
Ampicillin (or carbenicillin)	128 µg/ml	100 µg/ml	Resistant
Kanamycin	12.5-6.25 µg/ml (3.125µg/mL doesn't inhibit)	50 µg/ml	Sensitive	MIC info from Barrick Lab results (G.S.)
Spectinomycin	?	60 µg/ml	Sensitive	Spontaneous resistance
Streptomycin	6.25-12.5 µg/ml (escapes observed at 6.25µg/mL)	25 µg/ml	Sensitive	Barrick Lab results (G.S.)
Chloramphenicol	4.0 µg/ml	20 µg/ml	Sensitive	Spontaneous resistance
Gentamycin	1.3 µg/ml	5 µg/ml	Sensitive
Tetracycline	1.0 µg/ml	10 µg/ml	Sensitive
Rifampicin	0.6 µg/ml	100 µg/ml	Sensitive	Spontaneous resistance
Nalidixic acid (mutagen/causes DNA double strand breaks)	1.9 µg/ml (no growth at 24 hr, but growth at 48 hr) (1 µg/ml doesn't inhibit)	3.75 µg/ml	Sensitive	Barrick Lab results (G.S.)

Source: Gomez MJ, Neyfakh AA. (2006) Genes involved in intrinsic antibiotic resistance of Acinetobacter baylyi. Antimicrob Agents Chemother 50:3562–3567. DOI: 10.1128/AAC.00579-06

Plasmids

Common high-copy number E. coli plasmids do not work in ADP1.

Changed:

<
<

A synthetic plasmid (pBAV1K) and broad-host-range plasmid (RSF1010) have been used successfully. In our hands, pBAV1K is less reliable. It is also important to note that because plasmid DNA is linearizes when taken up through the competence apparatus, that transformation of plasmids can be significantly less efficient than transformations used to incorporate DNA directly in the chromosome.

>
>

A synthetic plasmid (pBAV1K) and broad-host-range plasmid (RSF1010) have been used successfully. In our hands, pBAV1K is less reliable. It is also important to note that because plasmid DNA is linearized when taken up through the competence apparatus, that transformation of plasmids can be significantly less efficient than transformations used to incorporate DNA directly in the chromosome.

Origin	Copy Number	Example
ColE1 (pUC, pBR322, etc.) type plasmids	Do not replicate	NA
pBAV1K	Low-medium copy number	pBAV1K-T5-gfp
RSF1010	Low-medium copy number	pBTK403

Revision 42022-03-18 - IsaacGifford

META TOPICPARENT	name="ProtocolList"

Acinetobacter baylyi ADP1 Overview

ADP1 Resources

Genome Sequences
- ADP1 type strain (GenBank Format)
  In general, use this genome as a reference when referring to coordinates of genes and features in the ADP1 genome. We added the locations of IS1236 elements to the linked sequence. Otherwise it is the same as the one on NCBI.
  - Original sequence of the ADP1 type strain at NCBI (GenBank: NC_005966.1)
  - BLAST a sequence against this genome at NCBI
- ADP1 Barrick Lab strain (GFF3 Format)
  Incorporates a small number of mutations found in the ADP1 strain that the Barrick lab originally obtained from Valérie de Crécy-Lagard. Use for analyzing NGS sequencing results.
- ADP1-ISx (GFF3 Format)
  Strain created in the Barrick lab with all IS1236 elements deleted.

Other Useful Sequences
- pBTK622
  Plasmid containing the tdk-kanR cassette used for genome editing.

Functional Genomics
- ADP1 pathways and gene functions (KEGG)
- ADP1 pathways and gene functions in (BioCyc)

Added:

>
>

Working with ADP1

Culture conditions
- LB is typically used when growing ADP1 in rich media and Minimal Succinate (ABMS) media is used when growing ADP1 in minimal media (recipes for both medias can be found here).
  - Media can be supplemented with antibiotics for selection of strains transformed with resistance markers as described below.
- ADP1 grows at both 30°C and 37°C. The Barrick Lab typically incubates ADP1 cultures at 30°C with shaking at 200 rpm.

Storage
- Cultures of ADP1 can be stored for a few days at 4°C.
- Glycerol or DMSO can be used as cryoprotectants when freezing cultures stocks (using the protocol here)

Genomic DNA extraction: The Barrick Lab uses a PureLink Genomic DNA Mini Kit for genomic DNA extractions. 30 minutes is sufficient incubation time at 55°C for lysing cultures of ADP1.

Protocols: the following protocols are commonly used when working with ADP1
- PCR Amplifying DNA.
- Golden Transformation Creating a construct for integration into the ADP1 genome with Golden Gate ligation and transforming it into ADP1. Constructs can be used for gene deletion, inserting DNA sequences, or introducing mutations.
- Growth curves Calculating growth rates using a plate reader.
- Competence assay Calculating transformation rates and confirming competence of ADP1 mutants.

Antibiotic Resistance

ADP1 is intrinsically resistant to beta-lactam antibiotics. Do not use plasmids/constructs that have beta-lactamase (bla) as the selectable marker gene for ampicillin (or carbenicillin) resistance. Most other common antibiotic selection markers can be used.

We most commonly use kanamycin and spectinomycin resistance markers. The rate of spontaneous mutations to KanR is undetectable. SpecR mutants can arise at a low frequency (~10⁻⁸ to 10⁻⁹) due to chromosomal mutations, so only use this marker for transformations that have a significantly higher frequency than this.

Antibiotic	MIC	Standard Conc.	Status	Notes
Ampicillin (or carbenicillin)	128 µg/ml	100 µg/ml	Resistant
Kanamycin	12.5-6.25 µg/ml (3.125µg/mL doesn't inhibit)	50 µg/ml	Sensitive	MIC info from Barrick Lab results (G.S.)
Spectinomycin	?	60 µg/ml	Sensitive	Spontaneous resistance
Streptomycin	6.25-12.5 µg/ml (escapes observed at 6.25µg/mL)	25 µg/ml	Sensitive	Barrick Lab results (G.S.)
Chloramphenicol	4.0 µg/ml	20 µg/ml	Sensitive	Spontaneous resistance
Gentamycin	1.3 µg/ml	5 µg/ml	Sensitive
Tetracycline	1.0 µg/ml	10 µg/ml	Sensitive
Rifampicin	0.6 µg/ml	100 µg/ml	Sensitive	Spontaneous resistance
Nalidixic acid (mutagen/causes DNA double strand breaks)	1.9 µg/ml (no growth at 24 hr, but growth at 48 hr) (1 µg/ml doesn't inhibit)	3.75 µg/ml	Sensitive	Barrick Lab results (G.S.)

Source: Gomez MJ, Neyfakh AA. (2006) Genes involved in intrinsic antibiotic resistance of Acinetobacter baylyi. Antimicrob Agents Chemother 50:3562–3567. DOI: 10.1128/AAC.00579-06

Plasmids

Common high-copy number E. coli plasmids do not work in ADP1.

A synthetic plasmid (pBAV1K) and broad-host-range plasmid (RSF1010) have been used successfully. In our hands, pBAV1K is less reliable. It is also important to note that because plasmid DNA is linearizes when taken up through the competence apparatus, that transformation of plasmids can be significantly less efficient than transformations used to incorporate DNA directly in the chromosome.

Origin	Copy Number	Example
ColE1 (pUC, pBR322, etc.) type plasmids	Do not replicate	NA
pBAV1K	Low-medium copy number	pBAV1K-T5-gfp
RSF1010	Low-medium copy number	pBTK403

Revision 32022-03-18 - JeffreyBarrick

META TOPICPARENT	name="ProtocolList"

Acinetobacter baylyi ADP1 Overview

ADP1 Resources

Genome Sequences

Changed:

<
<

- ADP1 Wild-Type (GenBank: NC_005966.1)
  In general, use this genome as a reference when referring to coordinates of genes and features in the ADP1 genome.
  - BLAST a sequence against the ADP1 (WT) genome (at NCBI)
- ADP1 Barrick (GenBank: NC_005966.1)
  Incorporates a small number of mutations found in the WT ADP1 strain that the Barrick lab originally obtained from Valérie de Crécy-Lagard. Use for analyzing NGS sequencing results.
- [[][ADP1-ISx]]
  Strain created in the Barrick lab with all IS1236 elements deleted. (NEEDS LINK)

>
>

- ADP1 type strain (GenBank Format)
  In general, use this genome as a reference when referring to coordinates of genes and features in the ADP1 genome. We added the locations of IS1236 elements to the linked sequence. Otherwise it is the same as the one on NCBI.
  - Original sequence of the ADP1 type strain at NCBI (GenBank: NC_005966.1)
  - BLAST a sequence against this genome at NCBI
- ADP1 Barrick Lab strain (GFF3 Format)
  Incorporates a small number of mutations found in the ADP1 strain that the Barrick lab originally obtained from Valérie de Crécy-Lagard. Use for analyzing NGS sequencing results.

Added:

>
>

- ADP1-ISx (GFF3 Format)
  Strain created in the Barrick lab with all IS1236 elements deleted.

Other Useful Sequences

Changed:

<
<

- tdk-kanR cassette used for genome editing

>
>

- pBTK622
  Plasmid containing the tdk-kanR cassette used for genome editing.

Functional Genomics
- ADP1 pathways and gene functions (KEGG)
- ADP1 pathways and gene functions in (BioCyc)

Antibiotic Resistance

ADP1 is intrinsically resistant to beta-lactam antibiotics. Do not use plasmids/constructs that have beta-lactamase (bla) as the selectable marker gene for ampicillin (or carbenicillin) resistance. Most other common antibiotic selection markers can be used.

We most commonly use kanamycin and spectinomycin resistance markers. The rate of spontaneous mutations to KanR is undetectable. SpecR mutants can arise at a low frequency (~10⁻⁸ to 10⁻⁹) due to chromosomal mutations, so only use this marker for transformations that have a significantly higher frequency than this.

Antibiotic	MIC	Standard Conc.	Status	Notes
Ampicillin (or carbenicillin)	128 µg/ml	100 µg/ml	Resistant
Kanamycin	12.5-6.25 µg/ml (3.125µg/mL doesn't inhibit)	50 µg/ml	Sensitive	MIC info from Barrick Lab results (G.S.)
Spectinomycin	?	60 µg/ml	Sensitive	Spontaneous resistance
Streptomycin	6.25-12.5 µg/ml (escapes observed at 6.25µg/mL)	25 µg/ml	Sensitive	Barrick Lab results (G.S.)
Chloramphenicol	4.0 µg/ml	20 µg/ml	Sensitive	Spontaneous resistance
Gentamycin	1.3 µg/ml	5 µg/ml	Sensitive
Tetracycline	1.0 µg/ml	10 µg/ml	Sensitive
Rifampicin	0.6 µg/ml	100 µg/ml	Sensitive	Spontaneous resistance
Nalidixic acid (mutagen/causes DNA double strand breaks)	1.9 µg/ml (no growth at 24 hr, but growth at 48 hr) (1 µg/ml doesn't inhibit)	3.75 µg/ml	Sensitive	Barrick Lab results (G.S.)

Source: Gomez MJ, Neyfakh AA. (2006) Genes involved in intrinsic antibiotic resistance of Acinetobacter baylyi. Antimicrob Agents Chemother 50:3562–3567. DOI: 10.1128/AAC.00579-06

Plasmids

Changed:

<
<

Common high-copy number E. coli plasmids do not work in ADP1. A synthetic plasmid (pBAV1K) and broad-host-range plasmid (RSF1010) have been used successfully. In our hands, pBAV1K is less reliable. It is also important to note that because plasmid DNA is linearizes when taken up through the competence apparatus, that transformation of plasmids can be significantly less efficient than transformations used to incorporate DNA directly in the chromosome.

>
>

Common high-copy number E. coli plasmids do not work in ADP1.

Added:

>
>

A synthetic plasmid (pBAV1K) and broad-host-range plasmid (RSF1010) have been used successfully. In our hands, pBAV1K is less reliable. It is also important to note that because plasmid DNA is linearizes when taken up through the competence apparatus, that transformation of plasmids can be significantly less efficient than transformations used to incorporate DNA directly in the chromosome.

Origin	Copy Number	Example
ColE1 (pUC, pBR322, etc.) type plasmids	Do not replicate	NA
pBAV1K	Low-medium copy number	pBAV1K-T5-gfp
RSF1010	Low-medium copy number	pBTK403

Revision 22022-03-18 - JeffreyBarrick

META TOPICPARENT	name="ProtocolList"

Acinetobacter baylyi ADP1 Overview

ADP1 Resources

Changed:

<
<

>
>

Genome Sequences
- ADP1 Wild-Type (GenBank: NC_005966.1)
  In general, use this genome as a reference when referring to coordinates of genes and features in the ADP1 genome.

Added:

>
>

- - BLAST a sequence against the ADP1 (WT) genome (at NCBI)
- ADP1 Barrick (GenBank: NC_005966.1)
  Incorporates a small number of mutations found in the WT ADP1 strain that the Barrick lab originally obtained from Valérie de Crécy-Lagard. Use for analyzing NGS sequencing results.
- [[][ADP1-ISx]]
  Strain created in the Barrick lab with all IS1236 elements deleted. (NEEDS LINK)

Changed:

<
<

| Quick Reference

>
>

Other Useful Sequences

Added:

>
>

- tdk-kanR cassette used for genome editing

Functional Genomics
- ADP1 pathways and gene functions (KEGG)
- ADP1 pathways and gene functions in (BioCyc)

Antibiotic Resistance

Added:

>
>

ADP1 is intrinsically resistant to beta-lactam antibiotics. Do not use plasmids/constructs that have beta-lactamase (bla) as the selectable marker gene for ampicillin (or carbenicillin) resistance. Most other common antibiotic selection markers can be used.

We most commonly use kanamycin and spectinomycin resistance markers. The rate of spontaneous mutations to KanR is undetectable. SpecR mutants can arise at a low frequency (~10⁻⁸ to 10⁻⁹) due to chromosomal mutations, so only use this marker for transformations that have a significantly higher frequency than this.

Antibiotic	MIC	Standard Conc.	Status	Notes
Ampicillin (or carbenicillin)	128 µg/ml	100 µg/ml	Resistant

Changed:

<
<

Spectinomycin

?

60 µg/ml

Sensitive

>
>

Kanamycin

12.5-6.25 µg/ml (3.125µg/mL doesn't inhibit)

50 µg/ml

Sensitive

MIC info from Barrick Lab results (G.S.)

Added:

>
>

Spectinomycin

?

60 µg/ml

Sensitive

Spontaneous resistance

Streptomycin

6.25-12.5 µg/ml (escapes observed at 6.25µg/mL)

25 µg/ml

Sensitive

Barrick Lab results (G.S.)

Deleted:

<
<

Kanamycin

12.5-6.25µg/ml (3.125µg/mL doesn't inhibit)

50 µg/ml

Sensitive

MIC info from Barrick Lab results (G.S.)

Chloramphenicol	4.0 µg/ml	20 µg/ml	Sensitive	Spontaneous resistance
Gentamycin	1.3 µg/ml	5 µg/ml	Sensitive
Tetracycline	1.0 µg/ml	10 µg/ml	Sensitive
Rifampicin	0.6 µg/ml	100 µg/ml	Sensitive	Spontaneous resistance

Changed:

<
<

Nalidixic acid (mutagen/causes DNA double strand breaks)

1.9µg/mL (no growth at 24hr, but growth at 48hr) (1µg/mL doesn't inhibit)

3.75 µg/ml

Sensitive

Barrick Lab results (G.S.)

>
>

Nalidixic acid (mutagen/causes DNA double strand breaks)

1.9 µg/ml (no growth at 24 hr, but growth at 48 hr) (1 µg/ml doesn't inhibit)

3.75 µg/ml

Sensitive

Barrick Lab results (G.S.)

Changed:

<
<

Source: Gomez MJ, Neyfakh AA. (2006) Genes involved in intrinsic antibiotic resistance of Acinetobacter baylyi. Antimicrob Agents Chemother 50:3562–3567. DOI: 10.1128/AAC.00579-06

>
>

Source: Gomez MJ, Neyfakh AA. (2006) Genes involved in intrinsic antibiotic resistance of Acinetobacter baylyi. Antimicrob Agents Chemother 50:3562–3567. DOI: 10.1128/AAC.00579-06

Plasmids

Changed:

<
<

Genome Resources: BLAST | BioCyc | Quick Reference

>
>

Common high-copy number E. coli plasmids do not work in ADP1. A synthetic plasmid (pBAV1K) and broad-host-range plasmid (RSF1010) have been used successfully. In our hands, pBAV1K is less reliable. It is also important to note that because plasmid DNA is linearizes when taken up through the competence apparatus, that transformation of plasmids can be significantly less efficient than transformations used to incorporate DNA directly in the chromosome.

Added:

>
>

Origin	Copy Number	Example
ColE1 (pUC, pBR322, etc.) type plasmids	Do not replicate	NA
pBAV1K	Low-medium copy number	pBAV1K-T5-gfp
RSF1010	Low-medium copy number	pBTK403

Revision 12022-03-17 - JeffreyBarrick

META TOPICPARENT	name="ProtocolList"

Acinetobacter baylyi ADP1 Overview

ADP1 Resources

| Quick Reference

Antibiotic Resistance

Antibiotic	MIC	Standard Conc.	Status	Notes
Ampicillin (or carbenicillin)	128 µg/ml	100 µg/ml	Resistant
Spectinomycin	?	60 µg/ml	Sensitive
Streptomycin	6.25-12.5 µg/ml (escapes observed at 6.25µg/mL)	25 µg/ml	Sensitive	Barrick Lab results (G.S.)
Kanamycin	12.5-6.25µg/ml (3.125µg/mL doesn't inhibit)	50 µg/ml	Sensitive	MIC info from Barrick Lab results (G.S.)
Chloramphenicol	4.0 µg/ml	20 µg/ml	Sensitive	Spontaneous resistance
Gentamycin	1.3 µg/ml	5 µg/ml	Sensitive
Tetracycline	1.0 µg/ml	10 µg/ml	Sensitive
Rifampicin	0.6 µg/ml	100 µg/ml	Sensitive	Spontaneous resistance
Nalidixic acid (mutagen/causes DNA double strand breaks)	1.9µg/mL (no growth at 24hr, but growth at 48hr) (1µg/mL doesn't inhibit)	3.75 µg/ml	Sensitive	Barrick Lab results (G.S.)

Source: Gomez MJ, Neyfakh AA. (2006) Genes involved in intrinsic antibiotic resistance of Acinetobacter baylyi. Antimicrob Agents Chemother 50:3562–3567. DOI: 10.1128/AAC.00579-06

Plasmids

Genome Resources: BLAST | BioCyc | Quick Reference

Difference: ProtocolsAcinetobacterBaylyiADP1 (1 vs. 6)

Revision 62022-03-22 - IsaacGifford

Acinetobacter baylyi ADP1 Overview

ADP1 Resources

Working with ADP1

Antibiotic Resistance

Plasmids

Revision 52022-03-22 - JeffreyBarrick

Acinetobacter baylyi ADP1 Overview

ADP1 Resources

Working with ADP1

Antibiotic Resistance

Plasmids

Revision 42022-03-18 - IsaacGifford

Acinetobacter baylyi ADP1 Overview

ADP1 Resources

Working with ADP1

Antibiotic Resistance

Plasmids

Revision 32022-03-18 - JeffreyBarrick

Acinetobacter baylyi ADP1 Overview

ADP1 Resources

Antibiotic Resistance

Plasmids

Revision 22022-03-18 - JeffreyBarrick

Acinetobacter baylyi ADP1 Overview

ADP1 Resources

Antibiotic Resistance

Plasmids

Revision 12022-03-17 - JeffreyBarrick

Acinetobacter baylyi ADP1 Overview

ADP1 Resources

Antibiotic Resistance

Plasmids