Source:
Generated By: https://synbiohub.org/public/igem/igem2sbol/1
Created by: Lorenzo Pasotti
Date created: 2016-01-02 12:00:00
Date modified: 2016-01-02 08:10:54
pSC101ts origin with SpeI removed
| Types | DnaRegion |
| Roles | DNA sequence_feature |
| Sequences | BBa_J107112_sequence (Version 1) |
Description
Temperature-sensitive pSC101 replication origin without SpeI site. This part was used to design the pBBknock BioBrick vector for allelic replacement (BBa_J107077).Notes
The origin has been retrieved from pAH123 vector [GenBank: AY048726]. The sequence was extended to the upstream NcoI restriction site and to the downstream stop codon of the ampicillin resistance gene in pAH123. We also modified the origin region by removing the SpeI site, according to the modification present in the available sequence of pSC101 (BBa_I50042), to ensure that the nucleotide changes do not affect the replication origin functioning, e.g., in the widely used pSB4C5 vector.The sequence of pAH123 was obtained via DNA sequencing, since its submitted DNA sequence [GenBank: AY048726] contains errors, e.g., in the temperature-sensitive mutation. The read sequence was then modified to remove SpeI site.