Basic Vector Information
- Vector Name:
- pSyn_Mz_Mj_sfGFP_190_200(amber-ochre)
- Antibiotic Resistance:
- Bleomycin
- Length:
- 3435 bp
- Type:
- Cloning vector
- Replication origin:
- p15A ori
- Source/Author:
- Fischer EC, Hashimoto K, Zhang Y, Feldman AW
pSyn_Mz_Mj_sfGFP_190_200(amber-ochre) vector Vector Map
pSyn_Mz_Mj_sfGFP_190_200(amber-ochre) vector Sequence
LOCUS 62056_20325 3435 bp DNA circular SYN 19-APR-2020
DEFINITION Cloning vector pSyn_Mz_Mj_sfGFP_190_200(amber-ochre), complete
sequence.
ACCESSION MN882184
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 3435)
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 3435)
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 3435)
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..3435
/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)"
CDS 1546..1635
/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 1686..1733
/label=T7 terminator
/note="transcription terminator for bacteriophage T7 RNA
polymerase"
regulatory 1763..1781
/note="promoter for bacteriophage T7 RNA polymerase; T7
promoter"
/regulatory_class="promoter"
protein_bind 1782..1806
/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 1818..1822
/label=5' proK leader
/note="5' proK leader"
misc_feature 1823..1894
/note="tRNA-Pyl; Methanosarcina mazei tRNAPyl(TTA)"
misc_feature 1895..1933
/label=3' proK trailer
/note="3' proK trailer"
regulatory 1934..1981
/note="transcription terminator for bacteriophage T7 RNA
polymerase; T7 terminator"
/regulatory_class="terminator"
regulatory 2028..2046
/note="promoter for bacteriophage T7 RNA polymerase; T7
promoter"
/regulatory_class="promoter"
protein_bind 2047..2071
/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 2083..2087
/label=5' proK leader
/note="5' proK leader"
misc_feature 2088..2164
/label=Methanocaldococcus jannaschii tRNApAzF(CTA)
/note="Methanocaldococcus jannaschii tRNApAzF(CTA)"
misc_feature 2165..2203
/label=3' proK trailer
/note="3' proK trailer"
regulatory 2204..2251
/note="transcription terminator for bacteriophage T7 RNA
polymerase; T7 terminator"
/regulatory_class="terminator"
regulatory 2289..2395
/label=proK promoter
/note="proK promoter"
/regulatory_class="promoter"
misc_RNA 2416..2491
/label=gRNA scaffold
/note="guide RNA scaffold for the Streptococcus pyogenes
CRISPR/Cas9 system"
regulatory 2499..2534
/label=proK terminator
/note="proK terminator"
/regulatory_class="terminator"
regulatory 2547..2652
/label=proK promoter
/note="proK promoter"
/regulatory_class="promoter"
misc_RNA 2671..2742
/note="guide RNA scaffold for the CRISPR/Cas9 system; gRNA
scaffold"
regulatory 2750..2785
/label=proK terminator
/note="proK terminator"
/regulatory_class="terminator"
promoter 2819..2866
/label=EM7 promoter
/note="synthetic bacterial promoter"
CDS 2885..3256
/label=BleoR
/note="antibiotic-binding protein"
terminator 3271..3365
/label=lambda t0 terminator
/note="transcription terminator from phage lambda"
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