AF Hadidi, United States Department of Agriculture
Vector
Name of construct: pSP65:pUAS14#4
Excise insert: EcoRI
Size of construct (kb): 3.3
Markers: ampR
Name of vector: pSP65
Intact vector size: 3.010
Type of vector: plasmid Vector end: EcoRI
Cloning sites: HindIII PstI SalI AccI HincII XbaI BamHI AvaI SmaI SacI
EcoRI
Polylinker sites: HindIII PstI SalI AccI HincII XbaI BamHI AvaI SmaI SacI
EcoRI
Host range:Escherichia coli
Features (with orientation and position when available):
enhancer: none
marker(s): ampR
promoter: SP6
replicon: pMB1
terminator: none
Insert
Genome: apple scar skin viroid (ASSV)
Genomic copy number: unique
Contains complete coding sequence?: U
Type of DNA: cDNA
Insert end: EcoRI
Insert end: EcoRI
Insert size (kb): 0.274
Cross references: DNA Seq. Acc.: M36646
Insert Size (kb)
0.274
Media
ATCC® Medium 1227: LB Medium (ATCC medium 1065) with 50 mcg/ml ampicillin
Biosafety Level
1
Biosafety classification is based on U.S. Public Health Service Guidelines, it is the responsibility of the customer to ensure that their facilities comply with biosafety regulations for their own country.
Comments
Restriction digests of the clone give the following sizes (kb): EcoRI--3.0, 0.3; PstI--3.3; BglI--3.3; HindIII--3.3.
Derived from nucleotides 286 to 228 in the sequence of the circular viroid.
If the plasmid is linearized with PstI or HindIII, transcription with SP6 polymerase will generate the complement (minus strand).
Classification
Aspcaviroid
References
Hashimoto J, Koganezawa H. Nucleotide sequence and secondary structure of apple scar skin viroid. Nucleic Acids Res. 15: 7045-7051, 1987. PubMed: 3658673