The complete nucleotide sequences of the SacBII Kan domain of the P1 pAD10-SacBII cloning vector and three cosmid cloning vectors: pTCF, svPHEP, and LAWRIST16.
The complete nucleotide sequence of the 16,009-bp SacBII Kan domain of the P1 pAD10-SacBII cloning vector and the sequences of three cosmid cloning vectors, pTCF (7941 bp), svPHEP (9201 bp), and LAWRIST16 (5194 bp) have been determined. A modified diatomaceous earth (Prep-A-Gene)-based procedure, which rapidly yields highly supercoiled double-stranded DNA from recombinant P1 and cosmid clones suitable for generating shotgun libraries, also has been developed. The isolated recombinant DNAs were physically sheared to generate 1- to 2-kb fragments that then were blunt-ended and subcloned into double-stranded pUC-based sequencing vectors. The double-stranded sequencing templates were isolated by an alkaline lysis method and subjected to Taq polymerase catalyzed fluorescent end-labeled primer cycle sequencing. After shotgun sequence assembly, contig gaps were closed and ambiguities were resolved via Sequenase catalyzed fluorescent dye-terminator sequencing.
Pan
H Q
HQ
Department of Chemistry and Biochemistry, University of Oklahoma, Norman 73019.
Wang
Y P
YP
Chissoe
S L
SL
Bodenteich
A
A
Wang
Z
Z
Iyer
K
K
Clifton
S W
SW
Crabtree
J S
JS
Roe
B A
BA
D017101:Bacteriophage P1 / Q000235:genetics*; D002892:Chromosomes, Human, Pair 22; D002899:Chromosomes, Human, Pair 9; D003001:Cloning, Molecular; D003360:Cosmids / Q000235:genetics*; D004247:DNA / Q000235:genetics / Q000302:isolation & purification; D004274:DNA, Recombinant / Q000235:genetics / Q000302:isolation & purification; D004278:DNA, Superhelical / Q000235:genetics / Q000302:isolation & purification; D004279:DNA, Viral / Q000235:genetics / Q000302:isolation & purification; D004259:DNA-Directed DNA Polymerase; D007692:Diatomaceous Earth; D016313:Genes, abl; D005822:Genetic Vectors / Q000235:genetics*; D006602:Hexosyltransferases; D006801:Humans; D007613:Kanamycin Resistance / Q000235:genetics*; D017422:Sequence Analysis, DNA / Q000379:methods*
D004274:DNA, Recombinant; D004278:DNA, Superhelical; D004279:DNA, Viral; D007692:Diatomaceous Earth; D004247:DNA; D006602:Hexosyltransferases; C021513:levansucrase; C034970:bacteriophage T7 induced DNA polymerase; D004259:DNA-Directed DNA Polymerase