Basic Vector Information
- Vector Name:
- pRED56-MD8-NY2
- Antibiotic Resistance:
- Ampicillin
- Length:
- 8258 bp
- Type:
- Gateway recycling vector
- Replication origin:
- ori
- Source/Author:
- Saisho T, Shibahara K, Nakao A, Tanaka Y, Nishimura K, Kimura T, Nakagawa T.
pRED56-MD8-NY2 vector Map
pRED56-MD8-NY2 vector Sequence
LOCUS 40924_36563 8258 bp DNA circular SYN 18-DEC-2018
DEFINITION Gateway recycling vector pRED56-MD8-NY2 DNA, complete sequence.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 8258)
AUTHORS Saisho T, Shibahara K, Nakao A, Tanaka Y, Nishimura K, Kimura T,
Nakagawa T.
TITLE Improved Gateway Recycling Cloning System for Multigene Construction
JOURNAL Unpublished
REFERENCE 2 (bases 1 to 8258)
AUTHORS Saisho T, Shibahara K, Nakao A, Tanaka Y, Nishimura K, Kimura T,
Nakagawa T.
TITLE Direct Submission
JOURNAL Submitted (02-NOV-2015) Contact:Tsuyoshi Nakagawa Shimane
University, Interdisciplinary Center for Science Research; 1060
Nishikawatsu, Matsue, Shimane 690-8504, Japan
REFERENCE 3 (bases 1 to 8258)
TITLE Direct Submission
REFERENCE 4 (bases 1 to 8258)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName:
"Unpublished"
COMMENT SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
(02-NOV-2015) Contact:Tsuyoshi Nakagawa Shimane University,
Interdisciplinary Center for Science Research; 1060 Nishikawatsu,
Matsue, Shimane 690-8504, Japan"
COMMENT SGRef: number: 3; type: "Journal Article"
COMMENT constructed using pDONR201.
FEATURES Location/Qualifiers
source 1..8258
/mol_type="other DNA"
/organism="synthetic DNA construct"
protein_bind 28..127
/label=attL1
/note="recombination site for the Gateway(R) LR reaction"
misc_feature 674..1071
/label=MD8
/note="MD8"
protein_bind 1109..1233
/label=attR1
/note="recombination site for the Gateway(R) LR reaction"
promoter 1270..1300
/label=lac UV5 promoter
/note="E. coli lac promoter with an 'up' mutation"
CDS 1354..2010
/codon_start=1
/label=CmR
/note="chloramphenicol acetyltransferase"
/translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
CDS 2355..2657
/codon_start=1
/label=ccdB
/note="CcdB, a bacterial toxin that poisons DNA gyrase"
/translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
VSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
protein_bind complement(2701..2825)
/label=attR2
/note="recombination site for the Gateway(R) LR reaction"
CDS 2836..3354
/codon_start=1
/label=VN173
/note="N-terminal fragment of mVenus for use in bimolecular
fluorescence complementation (BiFC) (Kodama and Hu, 2010)"
/translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
KFICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
VNFKIRHNIE"
terminator 3382..3634
/label=NOS terminator
/note="nopaline synthase terminator and poly(A) signal"
protein_bind 3843..3967
/label=attR4
/note="recombination site for the Gateway(R) LR reaction"
promoter 3992..4022
/label=lac UV5 promoter
/note="E. coli lac promoter with an ""up"" mutation"
misc_feature 4294..4334
/label=rare cutter sites
/note="rare cutter sites"
CDS complement(4426..5283)
/codon_start=1
/label=AmpR
/note="beta-lactamase"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
promoter complement(5284..5388)
/label=AmpR promoter
protein_bind complement(5685..5808)
/label=attR3
/note="recombination site for the Gateway(R) LR reaction"
protein_bind complement(6134..6229)
/label=attL6
/note="recombination site for the Gateway(R) LR reaction"
CDS 6353..7159
/codon_start=1
/label=KanR
/note="aminoglycoside phosphotransferase"
/translation="MSHIQRETSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP
DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA
FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
rep_origin 7305..7893
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
terminator complement(8028..8055)
/label=rrnB T2 terminator
/note="transcription terminator T2 from the E. coli rrnB
gene"
terminator complement(8147..8233)
/label=rrnB T1 terminator
/note="transcription terminator T1 from the E. coli rrnB
gene"
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