Application of vectors

Which antibiotic(s) can we use for growing the constructs?

The proper antibiotic is mentioned for all plasmids. The following concentrations are used:

Ampicillin (or Carbenicillin)     100 mg L-1

Kanamycin                              50 mg L-1

Spectinomycin                        100 mg L-1

Gentamicin                              25 mg L-1

Chloramphenicol                     30 mg L-1

 

Generally, binary vectors have spectinomycin resistance and high-copy plasmids are ampicillin resistant. Destination plasmids usually have a ccdB/CMR marker and so are also chloramphenicol resistant.

Why are my restriction digest/ sequencing results not compatible with the corresponding maps on this webpage?

We have sequenced most parts of the constructs, but unfortunately, there may still be some sequence errors in some vectors. Please do alert us if you find any discrepancies.

Where can we buy the enzymes needed for the LR recombination reaction?

You can order recombinase enzymes from Invitrogen.

Where can we get pDONR vectors?

You should order these pDONR vectors from Invitrogen. Some pDONR vectors are not in the Invitrogen catalog. However, Invitrogen is currently providing pDONR P3-P4 and pDONR P5-P6 as a custom service. The price is $500 USD for both vectors and they are only available as a pair. They provide 10 µg’s of DNA, maps and Vector NTI files for both vectors. To request these pDONR vectors, contact custom.services@invitrogen.com.

It is also possible to order publicly-available pDONR vectors from GeneCorner 

How can we design primers for N-terminal or C-terminal fusion to fluorescence tags?

C-terminal fusion

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N-terminal fusion

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What is the bacterial host strain of the plasmids?

The host strain of all plasmids is E. coli DB3.1 or ccdB survival cells from Invitrogen. Many plasmids contain the bacterial ccdB gene, which encodes an anti-DNA gyrase protein. The E. coli strain DB3.1 contains a mutation in the DNA gyrase gene so that the ccdB protein is unable to bind the mutant DNA Gyrase protein. In this strain the DNA is replicated normally and colonies grow.​

DB3.1 can be acquired from GeneCorner.

Website

I cannot see the Genbank files when I click on the 'DOWNLOAD .GB FILE' link.

We cannot fully control how Genbank files are displayed by your web browser. To properly download these files, you will have to right-click the link and then select 'save file as'. To view these files, you will need an installation of your favourite plasmid editing software (Geneious, CLC, Vector NTI,...) and then import the Genbank files into the program.

Orders

How are the vectors shipped?

The vectors are shipped as bacterial stab cultures.

Inside Europe, we send our parcels by first-class priority mail. Orders to destinations outside of Europe are sent by Federal Express.

How much does one vector cost?

The shipping and handling fee for each individual vector is €75. We recommend that you order at least 2 vectors for a minimum value of €150. You can order a single vector, but please note that this incurs a handling and shipping surcharge fee of €75. Starting from 2 vectors, this surcharge is waived. Belgian institutes and European Union institutes that have no VAT number must also pay 21% VAT.

How can I make sure my order will be allowed in by my country's customs?

For certain countries, we need additional data for customs clearance:

  • Australia and New Zealand require an AQUIS import permit granted to your lab by the relevant government.
  • Brazil requires a valid Brazillian company tax ID or CNPJ.

If your country requires this additional customs information, then please send it as soon as possible to the Gateway Vectors Shipping Staff.

Ordering issues

Questions concerning invoices

If you select bank transfer, you will be able to generate a pro forma invoice for your accounting department. An actual invoice is then sent after receipt of payment in Multisafepay.

Issues with bank transfers

When you want to pay via bank transfer, please make sure you follow the Multisafepay instructions that you have received in an e-mail.

Definitely double-check that you used the correct reference number, as this is the most common reason why bank transfers fail. Also make sure that the payment was transferred to the Multisafepay account and NOT the VIB account.

In these cases, the system won't recognize which order this payment is for, and the money will be automatically refunded.

Issues with VISA

You want to pay with a Visa card and you receive the following error: "Declined, not verified - Merchant only accepts 3DS verified cards".

When this happens, you will have to contact Visa yourself to help you. Most likely there is a security issue with your card, so we won't be able to help you with this.

Sequence issues

We are in the process of fully sequencing and updating our plasmids, especially the older Gateway vectors, as we are aware that there are some SNP's present.

Please do alert us if you find any other discrepancies or if you experience any problems with our plasmids.

Questions not answered here

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