Can I split the homologous 15-nt overlap between the insert and vector, or adjacent inserts?

Can I split the homologous 15-nt overlap between the insert and vector, or adjacent inserts?

Yes. The homologous 15-nt overlap can be split between adjacent DNA fragments. However, splitting the overlap between an insert and vector can only be done if the vector is linearized via inverse PCR.
Primer design for this option is not facilitated by the online Primer Design Tool.

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