Cloning vectors and vector components
First Claim
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1. A composition comprising a vector component, wherein said vector component comprises:
- i) first and second free ends;
ii) a selectable marker region, iii) an origin of replication iv) a first transcriptional terminator between said first free end and said selectable marker region, v) a second transcriptional terminator between said second free end and said origin of replication, and vi) a third transcriptional terminator between said origin of replication and said selectable marker region, and wherein said vector component is configured to form a circular recombinant vector when said first and second free ends are attached to an insert sequence.
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Abstract
The present invention relates to systems, methods, and compositions for cloning and sequencing insert nucleic acid sequences. In particular, the present invention provides vectors and vector components configured for multiplex cloning, multiplex sequencing, and fixed orientation cloning. The present invention also provides vectors and vector components that allow insert sequences that are deleterious to a host cell to be successfully cloned.
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Citations
15 Claims
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1. A composition comprising a vector component, wherein said vector component comprises:
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i) first and second free ends;
ii) a selectable marker region, iii) an origin of replication iv) a first transcriptional terminator between said first free end and said selectable marker region, v) a second transcriptional terminator between said second free end and said origin of replication, and vi) a third transcriptional terminator between said origin of replication and said selectable marker region, and wherein said vector component is configured to form a circular recombinant vector when said first and second free ends are attached to an insert sequence. - View Dependent Claims (2, 3, 4, 5, 6)
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7. A composition comprising a circular vector, wherein said circular vector comprises:
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a) a nucleic acid sequence comprising;
i) first and second ends, ii) a selectable marker region, iii) a first transcriptional terminator between said first end and said selectable marker region, and iv) a second transcriptional terminator between said second end and said selectable marker region; and
b) an insert sequence, wherein said insert sequence is attached to said first and second ends, and wherein said insert sequence comprises a base composition that is at least 65% A and T. - View Dependent Claims (8, 9, 10, 11, 12, 13)
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14. A composition comprising a vector component, wherein said vector component comprises:
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i) first and second free ends;
ii) a selectable marker region comprising a selectable marker sequence, wherein said selectable marker sequence comprises a first promoter, iii) a first transcriptional terminator between said first free end and said selectable marker region;
iv) a second transcriptional terminator between said second free end and said selectable marker region, and v) a third transcriptional terminator between said second free end and said selectable marker region, and wherein no other promoters are present in said vector component except said first promoter and said vector component is configured to form a circular recombinant vector when said first and second free ends are attached to an insert sequence. - View Dependent Claims (15)
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Specification