Basic Vector Information
pDONR221 is a cloning vector, with attP1 and attP2 sites and a kanamycin resistance marker.
- Vector Name:
- pDONR221
- Antibiotic Resistance:
- Chloramphenicol
- Length:
- 4761 bp
- Type:
- Gateway Cloning Vectors
- Replication origin:
- ori
- Source/Author:
- Invitrogen (Life Technologies)
- Copy Number:
- High copy number
- 5' Primer:
- M13 fwd
- 3' Primer:
- M13 rev
pDONR221 vector Map
References
- Kropp KN, Schäufele TJ, Fatho M, Volkmar M, Conradi R, Theobald M, Wölfel T, Wölfel C. A bicistronic vector backbone for rapid seamless cloning and chimerization of αβT-cell receptor sequences. PLoS One. 2020 Sep 9;15(9):e0238875.
pDONR221 vector Sequence
LOCUS Exported 4761 bp DNA circular SYN 13-DEC-2024
DEFINITION synthetic circular DNA
ACCESSION .
VERSION .
KEYWORDS pDONR221
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 4761)
AUTHORS 11111111
TITLE Direct Submission
FEATURES Location/Qualifiers
source 1..4761
/mol_type="other DNA"
/organism="synthetic DNA construct"
terminator 268..295
/label=rrnB T2 terminator
/note="transcription terminator T2 from the E. coli rrnB
gene"
terminator 387..473
/gene="Escherichia coli rrnB"
/label=rrnB T1 terminator
/note="transcription terminator T1 from the E. coli rrnB
gene"
primer_bind 537..553
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
protein_bind 570..801
/gene="mutant version of attP"
/label=attP1
/bound_moiety="BP Clonase(TM)"
/note="recombination site for the Gateway(R) BP reaction
(pDONR(TM)221 version)"
CDS complement(1197..1502)
/codon_start=1
/gene="ccdB"
/product="CcdB, a bacterial toxin that poisons DNA gyrase"
/label=ccdB
/note="Plasmids containing the ccdB gene cannot be
propagated in standard E. coli strains."
/translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
VSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
CDS complement(1852..2505)
/codon_start=1
/gene="cat"
/product="chloramphenicol acetyltransferase"
/label=CmR
/note="confers resistance to chloramphenicol"
/translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGG"
promoter complement(2506..2608)
/label=cat promoter
/note="promoter of the E. coli cat gene"
protein_bind complement(2753..2984)
/gene="mutant version of attP"
/label=attP2
/bound_moiety="BP Clonase(TM)"
/note="recombination site for the Gateway(R) BP reaction
(pDONR(TM)221 version)"
promoter complement(3003..3021)
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
primer_bind complement(3026..3042)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
CDS 3155..3964
/codon_start=1
/gene="aph(3')-Ia"
/product="aminoglycoside phosphotransferase"
/label=KanR
/note="confers resistance to kanamycin in bacteria or G418
(Geneticin(R)) in eukaryotes"
/translation="MSHIQRETSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP
DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA
FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
rep_origin 4111..4699
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
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