Basic Vector Information
- Vector Name:
- Master_Architecture_RbsR_HQN
- Antibiotic Resistance:
- Kanamycin
- Length:
- 5228 bp
- Type:
- Cloning vector
- Replication origin:
- pSC101 ori
- Source/Author:
- Rondon RE, Groseclose TM, Short AE, Wilson CJ.
Master_Architecture_RbsR_HQN vector Vector Map
Plasmid Resuspension Protocol:
1. Centrifuge at 5,000×g for 5 min.
2. Carefully open the tube and add 20 μl of sterile water to dissolve the DNA.
3. Close the tube and incubate for 10 minutes at room temperature.
4. Briefly vortex the tube and then do a quick spin to concentrate the liquid at the bottom. Speed is less than 5000×g.
5.Store the plasmid at -20 ℃.
Master_Architecture_RbsR_HQN vector Sequence
LOCUS 62056_1540 5228 bp DNA circular SYN 03-DEC-2019
DEFINITION Cloning vector Master_Architecture_RbsR_HQN, complete sequence.
ACCESSION MN207947
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 5228)
AUTHORS Rondon RE, Groseclose TM, Short AE, Wilson CJ.
TITLE Transcriptional programming using engineered systems of
transcription factors and genetic architectures
JOURNAL Nat Commun 10 (1), 4784 (2019)
PUBMED 31636266
REFERENCE 2 (bases 1 to 5228)
AUTHORS Rondon RE, Groseclose T, Short A, Wilson CJ.
TITLE Direct Submission
JOURNAL Submitted (21-JUL-2019) Chemical and Biomolecular Engineering,
Georgia Institute of Technology, 950 Atlantic dr. NW, 5110E,
Atlanta, GA 30332, United States
REFERENCE 3 (bases 1 to 5228)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Nat
Commun"; date: "2019"; volume: "10"; issue: "1"; pages: "4784"
COMMENT SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
(21-JUL-2019) Chemical and Biomolecular Engineering, Georgia
Institute of Technology, 950 Atlantic dr. NW, 5110E, Atlanta, GA
30332, United States"
COMMENT ##Assembly-Data-START##
Sequencing Technology :: Sanger dideoxy sequencing
##Assembly-Data-END##
FEATURES Location/Qualifiers
source 1..5228
/mol_type="other DNA"
/organism="synthetic DNA construct"
regulatory 51..56
/label=trc promoter -35 hexamer TTGACA
/note="trc promoter -35 hexamer TTGACA"
/regulatory_class="promoter"
regulatory 57..73
/label=operator Ottg
/note="operator Ottg"
/regulatory_class="other"
regulatory 74..79
/label=trc promoter -10 hexamer TATAAT
/note="trc promoter -10 hexamer TATAAT"
/regulatory_class="promoter"
regulatory 95..114
/label=operator Oagg
/note="operator Oagg"
/regulatory_class="other"
regulatory 133..182
/label=RiboJ10
/note="RiboJ10"
/regulatory_class="insulator"
regulatory 224..235
/label=strong bacterial RBS
/note="strong bacterial RBS"
/regulatory_class="ribosome_binding_site"
CDS 242..955
/label=superfolder GFP
/note="GFP variant that folds robustly even when fused to
poorly folded proteins (Nager et al., 2011)"
terminator 970..1056
/label=rrnB T1 terminator
/note="transcription terminator T1 from the E. coli rrnB
gene"
CDS complement(1261..2208)
/label=Rep101
/note="RepA protein needed for replication with the pSC101
origin"
rep_origin complement(2256..2478)
/direction=LEFT
/label=pSC101 ori
/note="low-copy replication origin that requires the Rep101
protein"
regulatory 2565..2570
/label=pLL promoter -35 hexamer TTGACA
/note="pLL promoter -35 hexamer TTGACA"
/regulatory_class="promoter"
regulatory 2571..2587
/label=operator Osym
/note="operator Osym"
/regulatory_class="other"
regulatory 2588..2593
/label=pLL promoter -10 hexamer GATACT
/note="pLL promoter -10 hexamer GATACT"
/regulatory_class="promoter"
regulatory 2609..2628
/label=operator Otta
/note="operator Otta"
/regulatory_class="other"
misc_RNA 2646..2696
/label=sTRSV HHRz
/note="hammerhead ribozyme from the tobacco ringspot virus
satellite RNA (Khvorova et al., 2003)"
regulatory 2698..2719
/label=RiboJ
/note="RiboJ"
/regulatory_class="insulator"
regulatory 2724..2735
/label=strong bacterial RBS
/note="strong bacterial RBS"
/regulatory_class="ribosome_binding_site"
CDS 2742..3740
/codon_start=1
/transl_table=11
/product="RbsR HQN"
/label=RbsR HQN
/protein_id="QFU95565.1"
/translation="MKPVTLYDVAEYAGVSHQTVSNVVNQASHVSAKTREKVEAAMAEL
NYIPNRVAQQLAGKASHTIGMLITASTNPFYSELVRGVERSCFERGYSLVLCNTEGDEQ
RMNRNLETLMQKRVDGLLLLCTETHQPSREIMQRYPTVPTVMMDWAPFDGDSDLIQDNS
LLGGDLATQYLIDKGHTRIACITGPLDKTPARLRLEGYRAAMKRAGLNIPDGYEVTGDF
EFNGGFDAMRQLLSHPLRPQAVFTGNDAMAVGVYQALYQAELQVPQDIAVIGYDDIELA
SFMTPPLTTIHQPKDELGELAIDVLIHRITQPTLQQQRLQLTPILMERGSA"
terminator 3741..3772
/label=tonB terminator
/note="bidirectional E. coli tonB-P14 transcription
terminator"
terminator complement(4173..4267)
/label=lambda t0 terminator
/note="transcription terminator from phage lambda"
CDS complement(4301..5092)
/label=NeoR/KanR
/note="aminoglycoside phosphotransferase"
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