Basic Vector Information
pSC101 ori is a low-copy replication origin that requires the Rep101 protein, which is a temperature-sensitive protein. When bacteria containing a plasmid with this ori is cultured at 37℃, the plasmid will be lost.
- Vector Name:
- pKfloxANTSAsc-tsOri-pBR
- Antibiotic Resistance:
- Kanamycin
- Length:
- 4436 bp
- Type:
- Cloning vector
- Replication origin:
- pSC101 ori
- Source/Author:
- Yoon YG, Koob MD.
- Copy Number:
- Low Copy
- Growth Temperature:
- 30℃
pKfloxANTSAsc-tsOri-pBR vector Map
pKfloxANTSAsc-tsOri-pBR vector Sequence
LOCUS 40924_26664 4436 bp DNA circular SYN 18-DEC-2018
DEFINITION Cloning vector pKfloxANTSAsc-tsOri-pBR, complete sequence.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 4436)
AUTHORS Yoon YG, Koob MD.
TITLE Nonreplicating Intracellular Bacterial Vector for Conjugative DNA
Transfer into Mitochondria
JOURNAL Pharm. Res. (2012) In press
PUBMED 22350804
REFERENCE 2 (bases 1 to 4436)
AUTHORS Koob MD.
TITLE Direct Submission
JOURNAL Submitted (16-DEC-2011) Lab Medicine and Pathology, ITN and IHG,
University of Minnesota, 2101 6th St SE, WMBB, Minneapolis, MN
55455, USA
REFERENCE 3 (bases 1 to 4436)
TITLE Direct Submission
REFERENCE 4 (bases 1 to 4436)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Pharm. Res.
(2012) In press"
COMMENT SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
(16-DEC-2011) Lab Medicine and Pathology, ITN and IHG, University of
Minnesota, 2101 6th St SE, WMBB, Minneapolis, MN 55455, USA"
COMMENT SGRef: number: 3; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..4436
/mol_type="other DNA"
/organism="synthetic DNA construct"
terminator 55..301
/label=lambda tL3 terminator
/note="transcription terminator tL3 from phage lambda"
CDS complement(406..1353)
/codon_start=1
/label=Rep101
/note="RepA protein needed for replication with the pSC101
origin"
/translation="MSELVVFKANELAISRYDLTEHETKLILCCVALLNPTIENPTRKE
RTVSFTYNQYAQMMNISRENAYGVLAKATRELMTRTVEIRNPLVKGFEIFQWTNYAKFS
SEKLELVFSEEILPYLFQLKKFIKYNLEHVKSFENKYSMRIYEWLLKELTQKKTHKANI
EISLDEFKFMLMLENNYHEFKRLNQWVLKPISKDLNTYSNMKLVVDKRGRPTDTLIFQV
ELDRQMDLVTELENNQIKMNGDKIPTTITSDSYLHNGLRKTLHDALTAKIQLTSFEAKF
LSDMQSKYDLNGSFSWLTQKQRTTLENILAKYGRI"
rep_origin complement(1401..1623)
/direction=LEFT
/label=pSC101 ori
/note="low-copy replication origin that requires the Rep101
protein"
rep_origin complement(2345..2933)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
protein_bind 3073..3106
/label=loxP
/note="Cre-mediated recombination occurs in the 8-bp core
sequence (ATGTATGC) (Shaw et al., 2021)."
CDS complement(3127..3918)
/codon_start=1
/label=NeoR/KanR
/note="aminoglycoside phosphotransferase"
/translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
terminator 4160..4246
/label=rrnB T1 terminator
/note="transcription terminator T1 from the E. coli rrnB
gene"
terminator 4338..4365
/label=rrnB T2 terminator
/note="transcription terminator T2 from the E. coli rrnB
gene"
misc_feature complement(join(4383..4436,1..3))
/label=MCS
/note="pUC18/19 multiple cloning site"
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