Basic Vector Information
- Vector Name:
- pSyn_Mz_Mj_sfGFP_190-200(dest)
- Antibiotic Resistance:
- Bleomycin
- Length:
- 3323 bp
- Type:
- Cloning vector
- Replication origin:
- p15A ori
- Source/Author:
- Fischer EC, Hashimoto K, Zhang Y, Feldman AW
- Promoter:
- EM7
pSyn_Mz_Mj_sfGFP_190-200(dest) vector Vector Map
pSyn_Mz_Mj_sfGFP_190-200(dest) vector Sequence
LOCUS 62056_20320 3323 bp DNA circular SYN 19-APR-2020
DEFINITION Cloning vector pSyn_Mz_Mj_sfGFP_190-200(dest), complete sequence.
ACCESSION MN882187
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 3323)
AUTHORS Fischer EC, Hashimoto K, Zhang Y, Feldman AW, Dien VT, Karadeema RJ,
Adhikary R, Ledbetter MP, Krishnamurthy R, Romesberg FE.
TITLE New codons for efficient production of unnatural proteins in a
semi-synthetic organism
JOURNAL Nat. Chem. Biol. (2020) In press
REFERENCE 2 (bases 1 to 3323)
AUTHORS Fischer EC.
TITLE Direct Submission
JOURNAL Submitted (27-DEC-2019) Department of Chemistry, The Scripps
Research Institute, 10550 North Torrey Pines Road, La Jolla, CA
92037, USA
REFERENCE 3 (bases 1 to 3323)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Nat. Chem.
Biol. (2020) In press"
COMMENT SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
(27-DEC-2019) Department of Chemistry, The Scripps Research
Institute, 10550 North Torrey Pines Road, La Jolla, CA 92037, USA"
FEATURES Location/Qualifiers
source 1..3323
/mol_type="other DNA"
/organism="synthetic DNA construct"
rep_origin 48..593
/label=p15A ori
/note="Plasmids containing the medium-copy-number p15A
origin of replication can be propagated in E. coli cells
that contain a second plasmid with the ColE1 origin."
promoter 746..764
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
protein_bind 766..784
/label=tet operator
/note="bacterial operator O2 for the tetR and tetA genes"
RBS 799..821
/label=RBS
/note="efficient ribosome binding site from bacteriophage
T7 gene 10 (Olins and Rangwala, 1989)"
misc_feature 1379..1396
/label=D190-Y200 Golden Gate destination site
/note="D190-Y200 Golden Gate destination site"
CDS 1495..1518
/label=Strep-Tag II
/note="peptide that binds Strep-Tactin(R), an engineered
form
of streptavidin"
CDS 1555..1578
/label=Strep-Tag II
/note="peptide that binds Strep-Tactin(R), an engineered
form
of streptavidin"
terminator 1632..1679
/label=T7 terminator
/note="transcription terminator for bacteriophage T7 RNA
polymerase"
regulatory 1709..1727
/note="promoter for bacteriophage T7 RNA polymerase; T7
promoter"
/regulatory_class="promoter"
protein_bind 1728..1752
/label=lac operator
/note="The lac repressor binds to the lac operator to
inhibit transcription in E. coli. This inhibition can be
relieved by adding lactose or
isopropyl-beta-D-thiogalactopyranoside (IPTG)."
misc_feature 1764..1768
/label=5' proK leader
/note="5' proK leader"
misc_feature 1769..1816
/note="tRNA-Pyl; Methanosarcina mazei tRNAPyl(dest)"
misc_feature 1776..1793
/label=tRNAPyl Golden Gate destination site
/note="tRNAPyl Golden Gate destination site"
misc_feature 1817..1855
/label=3' proK trailer
/note="3' proK trailer"
regulatory 1856..1903
/note="transcription terminator for bacteriophage T7 RNA
polymerase; T7 terminator"
/regulatory_class="terminator"
regulatory 1950..1968
/note="promoter for bacteriophage T7 RNA polymerase; T7
promoter"
/regulatory_class="promoter"
protein_bind 1969..1993
/label=lacI binding site
/bound_moiety="lacI"
/note="lac repressor binds to the lac operator to inhibit
transcription in E. coli; lac operator"
misc_feature 2005..2009
/label=5' proK leader
/note="5' proK leader"
misc_feature 2010..2052
/label=Methanocaldococcus jannaschii tRNApAzF(dest)
/note="Methanocaldococcus jannaschii tRNApAzF(dest)"
misc_feature 2020..2037
/label=tRNApAzF Golden Gate destination site
/note="tRNApAzF Golden Gate destination site"
misc_feature 2053..2091
/label=3' proK trailer
/note="3' proK trailer"
regulatory 2092..2139
/note="transcription terminator for bacteriophage T7 RNA
polymerase; T7 terminator"
/regulatory_class="terminator"
regulatory 2177..2283
/label=proK promoter
/note="proK promoter"
/regulatory_class="promoter"
misc_RNA 2304..2379
/label=gRNA scaffold
/note="guide RNA scaffold for the Streptococcus pyogenes
CRISPR/Cas9 system"
regulatory 2387..2422
/label=proK terminator
/note="proK terminator"
/regulatory_class="terminator"
regulatory 2435..2540
/label=proK promoter
/note="proK promoter"
/regulatory_class="promoter"
misc_RNA 2559..2630
/note="guide RNA scaffold for the CRISPR/Cas9 system; gRNA
scaffold"
regulatory 2638..2673
/label=proK terminator
/note="proK terminator"
/regulatory_class="terminator"
promoter 2707..2754
/label=EM7 promoter
/note="synthetic bacterial promoter"
CDS 2773..3144
/label=BleoR
/note="antibiotic-binding protein"
terminator 3159..3253
/label=lambda t0 terminator
/note="transcription terminator from phage lambda"
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