Yes, I have made it myself several times using pTZ57R, pGEMT and pBlueScript.

Please see this paper: http://www.ncbi.nlm.nih.gov/pmc/articles/PMC333800/

In the paper, authors state that If you give equimolar amounts of dNTPs to Taq DNA polymerase, dATP will be preferably added. However, if you supply only dTTP, then Taq will add an extra T to the end of any blunt ended DNA fragment. Based on that you can prepare your own T-vector using any vector backbone of choice.

Here follow a detailed protocol (as I usually do): 

-1X Taq Buffer - final

-MgCl2 to 2.5 mM final concentration

-10 units of polimerase (yes, it is a lot)

-Blunt ended Vector (up to 5 micrograms)

-ultra pure water to 100 microliters.

Incubate for 1 to 2 hours at 72°C (test with your avaialble Taq)

Purify by silica column kit (or simply by precipitation) and quantify by nanodrop or any other method.

Easy and cheap!

Other option:

If you are amplifying a target gene using polymerases that leave blunt ends, you can easily improve your blunt end ligation by adding a few units of blunt end enzyme to your ligation system. In my hands, EcoRV and SmaI cuts very well in the 1X Thermo Fischer ligase buffer.

Link for the original idea: http://www.academicjournals.org/journal/AJB/article-full-text-pdf/7EEAD3525311

P.S: Ensure that there are no cut site for the blunt restriction enzyme of your choice in the target fragment to be cloned.

I hope things are clearer now.

All the best

PGEM-T is a linearized cloning vector that can not be multiplied. I am afraid you will have to buy it again and again.

Furthermore it has the two T overhangs that preclude its ligation ..... As Christophe said you will have to buy it

If you want to create your own T-vector for T/A cloning in your lab:

Researchers have tried to develop pGEM-T -like T-vector. They claimed that their created new version pGEM-FT (for T/A cloning) has a better performance then pGEM-T vector. The vector also has T7 and SP6 promoters. See the attached paper. You may consider to ask the plasmid from the group to try out in your lab.

1 - Pick a blue colony from your transformation plate;

2 - Purify the plasmid;

3 - Linearize the plasmid by EcoRV Cut;

4 - Add T-overhang by treating the Linearized plasmid with taq polimerase and dTTP in a PCR like reaction;

5 - Purify, quantify and you are ready to go.

@Marcelo,

Interesting answer.

1. The company (Promega) use similar way in producing pGEM-T vector. But they use  pGEM®-5Zf(+) as start material (see attachment).

2. Although seldom, self-ligation of pGEM-T has been reported and discussed on ResearchGate.

3. I am wondering, have you done that (in your post above) yourself? How 'efficient' is it to attach the T to the EcoRV-cut blunt-ends?

Yes, I have made it myself several times using pTZ57R, pGEMT and pBlueScript.

Please see this paper: http://www.ncbi.nlm.nih.gov/pmc/articles/PMC333800/

In the paper, authors state that If you give equimolar amounts of dNTPs to Taq DNA polymerase, dATP will be preferably added. However, if you supply only dTTP, then Taq will add an extra T to the end of any blunt ended DNA fragment. Based on that you can prepare your own T-vector using any vector backbone of choice.

Here follow a detailed protocol (as I usually do): 

-1X Taq Buffer - final

-MgCl2 to 2.5 mM final concentration

-10 units of polimerase (yes, it is a lot)

-Blunt ended Vector (up to 5 micrograms)

-ultra pure water to 100 microliters.

Incubate for 1 to 2 hours at 72°C (test with your avaialble Taq)

Purify by silica column kit (or simply by precipitation) and quantify by nanodrop or any other method.

Easy and cheap!

Other option:

If you are amplifying a target gene using polymerases that leave blunt ends, you can easily improve your blunt end ligation by adding a few units of blunt end enzyme to your ligation system. In my hands, EcoRV and SmaI cuts very well in the 1X Thermo Fischer ligase buffer.

Link for the original idea: http://www.academicjournals.org/journal/AJB/article-full-text-pdf/7EEAD3525311

P.S: Ensure that there are no cut site for the blunt restriction enzyme of your choice in the target fragment to be cloned.

I hope things are clearer now.

All the best

Marcelo, Thank you so much for your answers.

Marcelo, great answer though I don't see any concentration of dTTP listed in your protocol. How much of this do you typically add?

Hi Eli, I usually add 0.2 mM of dTTP for each reaction. I just have followed what is recommended for a typical PCR where you add 0.2 mM of mixed dNTPs. Certainly 0.2 mM of dTTP is way too much, considering that will be no DNA synthesis... However I did like that once, and had enough vector to clone for a long period of time...

Regards

good job bro