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Part:BBa_K533000:Design

Designed by: ZHANG Yunxiao   Group: iGEM11_Tsinghua   (2011-09-29)

p15A replication origin


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 3
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal AgeI site found at 89
    Illegal AgeI site found at 413
  • 1000
    COMPATIBLE WITH RFC[1000]


Design Notes

It is advisable to distribute the genetic elements onto different plasmids in the same cell. In this way, the size limit of a single plasmid can be avoided in part. Furthermore, combination of different plasmids can confer a flexibility that can hardly be seen when using one plasmid alone.

However, issues of compatibility frequently occur when multiple plasmids are used together. Plasmids in the same compatibility group cannot coexist in the same cell. The most commonly used pUC replication origin belongs to a compatibility group that covers pMB1 origin (pBR and its derivatives) and ColE origin, which precludes coexistence of many plasmids.

Granted, plasmids with replication origins that fall in other compatibility groups are myriad, but most of them have very low copy numbers and incapable of chloramphenicol amplification, making it rather difficult to do molecular experiments.

In this part, we cloned a replication origin derivative of p15A origin. It is different from the original one in that it contains several mutations, which increases the copy number. In this way, both the compatibility and the convenience for use are accommodated.

Source

Source: Cloned from plasmid pACYC184

References