How
to Find Clones of Interest
|
How
to Find Clones of Interest |
Definition of SARS(TWJ) ORFs:
| ORF ID | Position on the SARS Genome | Product | Protein ID | Note |
| orf1ab | join(265..13398,13398..21485) | product="polyprotein 1ab" | protein_id="BAC81360" | potential ribosome slippery site |
| S | 21492..25259 | product="S" | protein_id="BAC81362.1" | |
| orf3 | 25268..26092 | product="hypothetical protein" | protein_id="BAC81363.1" | |
| orf4 | 25689..26153 | product="hypothetical protein" | protein_id="BAC81364.1" | |
| E | 26117..26347 | product="E" | protein_id="BAC81365.1" | |
| M | 26398..27063 | product="M" | protein_id="BAC81366.1" | |
| orf7 | 27074..27196 | product="hypothetical protein" | protein_id="BAC81367.1" | note="a 2-base deletion (27169-27170delTA) causing a frameshift and introducing a premature translation stop codon" |
| orf8 | 27271..27639 | product="hypothetical protein" | protein_id="BAC81368.1" | |
| orf9 | 27636..27770 | product="hypothetical protein" | protein_id="BAC81369.1" | |
| orf10 | 27777..27896 | product="hypothetical protein" | protein_id="BAC81370.1" | |
| orf11 | 27862..28116 | product="hypothetical protein" | protein_id="BAC81371.1" | |
| N | 28118..29386 | product="nucleocapsid protein N" | protein_id="BAC81372.1" | |
| orf13 | 28128..28424 | product="hypothetical protein" | protein_id="BAC81373.1" |
Definition of pCR101-3:
Using PCR mutagenesis, the specific 29 base pairs (cctactggttaccaacctgaatggaatat) of GZ29-nt-Ins was inserted between t (27881) and a (27882) of TWJ cDNA region (27777-28116).
This clone
contains partially overlapped orf10 and orf11 genes, resulting in an artificial
BGI-PUP(GZ29-nt-Ins)-like
CDS construct.
SARS(TWJ) ORFs Clone List
Clone
name |
Sequence |
ORF
ID |
Insert size(bp) |
Cloning
vector |
PCR
primer (forward) |
PCR
primer (reverse) |
Strain |
Note |
| pCRSj-21 | Seq | S (1-3768) | 3768 | pDNR-dual | CCTCGAGATGTTTATTTTCTTATTATTTC | GTCTAGAAATGTGTAATGTAATTTGACACC | TWJ |
WT; Fusion of SalI in vector and XhoI in insert without regeneration of either site. |
| pCRSjN-1 | Seq | S (1-1929) | 1929 | pDNR-dual | CCTCGAGATGTTTATTTTCTTATTATTTC | ACGGAGGTTTTAGCCATAG | TWJ |
WT; 5'-half; Fusion of SalI in vector and XhoI in insert without regeneration of either site. |
| pCRSjC-8 | Seq | S (1924-3768) | 1842 | pDNR-dual | TTACACCTGGAACAAATGC | GTCTAGAAATGTGTAATGTAATTTGACACC | TWJ |
WT; 3'-half; without stop codon |
| pCR3-9 | Seq | orf3;X1 | 822 | pDNR-dual | GAAGTTATCAGTCGACATGGATTTGTTTATGAGATTTTTTACTC | ATGGTCTAGAAAGCTTCAAAGGCACGCTAGTAGTCGTCG |
TWJ |
WT; without stop codon |
| pCR4-35 | Seq | orf4;X2 | 465 | pDNR-dual | GAAGTTATCAGTCGACATGATGCCAACTACTTTGTTTGC | ATGGTCTAGAAAGCTTTTAACGTACCTGTTTCTTCCGAA |
TWJ |
WT; with stop codon |
| pCREj-2 | Seq | E | 231 | pDNR-dual | GAAGTTATCAGTCGACATGTACTCATTCGTTTCGGAAGA | ATGGTCTAGAAAGCTTGACCAGAAGATCAGGAACTCCTT
|
TWJ |
WT; without stop codon |
| pCRMj-2 | Seq | M | 663 | pDNR-dual | GAAGTTATCAGTCGACATGGCAGACAACGGTACTATTACC | ATGGTCTAGAAAGCTTCTGTACTAGCAAAGCAATATTGTCG | TWJ |
WT; without stop codon |
| pCR7j-1 | Seq | orf7;X3 | 189 | pDNR-dual | GAAGTTATCAGTCGACATGTTTCATCTTGTTGACTTCCAG |
ATGGTCTAGAAAGCTTTGGATAATCTAACTCCATAGGTTCTT |
TWJ |
WT; with stop codon |
| pCR8-33 | Seq | orf8;X4 | 369 | pDNR-dual | GAAGTTATCAGTCGACATGAAAATTATTCTCTTCCTGACATT | ATGGTCTAGAAAGCTTTCATTCTGTCTTTCTCTTAATGGTG |
TWJ |
WT; with stop codon |
| pCR9-10 | Seq | orf9 | 132 | pDNR-dual | GAAGTTATCAGTCGACATGTTTCATCTTGTTGACTTCCAG
|
ATGGTCTAGAAAGCTTAGACTTTGGTACAAGGTTC
|
TWJ |
WT; with stop codon |
| pCR10j-3 | Seq | orf10 | 117 | pDNR-dual | GAAGTTATCAGTCGACATGAAACTTCTCATTGTTTTGACTTG | ATGGTCTAGAAAGCTTGTGTTGTACCTTACAAGGATCTTCAA | TWJ |
WT; without stop codon |
| pCR11-1 | Seq | orf11;X5 | 252 | pDNR-dual | GAAGTTATCAGTCGACATGTGCTTGAAGATCCTTGTAAGG | ATGGTCTAGAAAGCTTATTTGTTCGTTTATTTAAAACAACAA | TWJ |
WT; without stop codon |
| pCRN-2 | Seq | N | 1266 | pDNR-dual | GAAGTTATCAGTCGACATGTCTGATAATGGACCCCAATC | ATGGTCTAGAAAGCTTTGCCTGAGTTGAATCAGCAGAAG | TWJ |
WT; without stop codon |
| pCR13-3 | Seq | orf13 | 294 | pDNR-dual | GAAGTTATCAGTCGACATGGACCCCAATCAAACCAA | ATGGTCTAGAAAGCTTTTTTGCCGTCACCACCACG |
TWJ |
WT; without stop codon |
| pCR14-18 | Seq | orf14 | 210 | pDNR-dual | GAAGTTATCAGTCGACATGCTGCCACCGTGCTACA | ATGGTCTAGAAAGCTTGCAGCAATAGCGCGAGGG |
TWJ |
WT; without stop codon |
pCR101-3
|
Seq | orf10+29bp+orf11 |
366 | pDNR-dual | GAAGTTATCAGTCGACATGAAACTTCTCATTGTTTTGACTTG | ATGGTCTAGAAAGCTTATTTGTTCGTTTATTTAAAACAACAA | GD01 |
WT; without stop codon;29 bp insertion between orf 10 and 11 |
*ORF-1：本中心有部分 ORF-1 片段殖株,若有特別需求,請洽本中心
E-Mail : libres@ym.edu.tw |
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