Basic Vector Information
- Vector Name:
- pSyn_Mz_Mj_Eser_sfGFP_151_190-200(dest)
- Antibiotic Resistance:
- Bleomycin
- Length:
- 3064 bp
- Type:
- Cloning vector
- Replication origin:
- p15A ori
- Source/Author:
- Fischer EC, Hashimoto K, Zhang Y, Feldman AW
pSyn_Mz_Mj_Eser_sfGFP_151_190-200(dest) vector Map
pSyn_Mz_Mj_Eser_sfGFP_151_190-200(dest) vector Sequence
LOCUS 62056_20315 3064 bp DNA circular SYN 19-APR-2020
DEFINITION Cloning vector pSyn_Mz_Mj_Eser_sfGFP_151_190-200(dest), complete
sequence.
ACCESSION MN882188
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 3064)
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 3064)
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 3064)
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..3064
/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 1248..1265
/label=Y151 Golden Gate destination site
/note="Y151 Golden Gate destination site"
misc_feature 1339..1356
/label=D190-Y200 Golden Gate destination site
/note="D190-Y200 Golden Gate destination site"
CDS 1449..1538
/label=Twin-Strep-tag
/note="two Strep-Tag(R)II moieties connected by a linker
for enhanced binding to Strep-Tactin(R), an engineered form
of streptavidin (Schmidt et al., 2013)"
terminator 1589..1636
/label=T7 terminator
/note="transcription terminator for bacteriophage T7 RNA
polymerase"
regulatory 1666..1684
/note="promoter for bacteriophage T7 RNA polymerase; T7
promoter"
/regulatory_class="promoter"
protein_bind 1685..1709
/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 1721..1725
/label=5' proK leader
/note="5' proK leader"
misc_feature 1726..1773
/note="tRNA-Pyl; Methanosarcina mazei tRNAPyl(dest)"
misc_feature 1733..1750
/label=tRNAPyl Golden Gate destination site
/note="tRNAPyl Golden Gate destination site"
misc_feature 1774..1812
/label=3' proK trailer
/note="3' proK trailer"
regulatory 1813..1860
/note="transcription terminator for bacteriophage T7 RNA
polymerase; T7 terminator"
/regulatory_class="terminator"
regulatory 1907..1925
/note="promoter for bacteriophage T7 RNA polymerase; T7
promoter"
/regulatory_class="promoter"
protein_bind 1926..1950
/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 1962..1966
/label=5' proK leader
/note="5' proK leader"
misc_feature 1967..2009
/note="tRNA; Methanocaldococcus jannaschii tRNApAzF(dest)"
misc_feature 1977..1994
/label=tRNApAzF Golden Gate destination site
/note="tRNApAzF Golden Gate destination site"
misc_feature 2010..2048
/label=3' proK trailer
/note="3' proK trailer"
regulatory 2049..2096
/note="transcription terminator for bacteriophage T7 RNA
polymerase; T7 terminator"
/regulatory_class="terminator"
regulatory 2181..2199
/note="promoter for bacteriophage T7 RNA polymerase; T7
promoter"
/regulatory_class="promoter"
protein_bind 2200..2217
/label=tetracycline repressor TetR binding site
/bound_moiety="tetracycline repressor TetR"
/note="bacterial operator O2 for the tetR and tetA genes;
tet operator"
misc_feature 2230..2236
/label=5' serT leader
/note="5' serT leader"
misc_feature 2237..2298
/label=Escherichia coli tRNASer(dest)
/note="Escherichia coli tRNASer(dest)"
misc_feature 2251..2268
/label=tRNASer Golden Gate destination site
/note="tRNASer Golden Gate destination site"
misc_feature 2299..2341
/label=3' serT trailer
/note="3' serT trailer"
regulatory 2342..2389
/note="transcription terminator for bacteriophage T7 RNA
polymerase; T7 terminator"
/regulatory_class="terminator"
promoter 2448..2495
/label=EM7 promoter
/note="synthetic bacterial promoter"
CDS 2514..2885
/label=BleoR
/note="antibiotic-binding protein"
terminator 2900..2994
/label=lambda t0 terminator
/note="transcription terminator from phage lambda"
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