pSyn_Mz_Mj_sfGFP_190_200(amber-ochre) vector (V016163)

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 Map

pSyn_Mz_Mj_sfGFP_190_200(amber-ochre)3435 bp6001200180024003000p15A oriT7 promotertet operatorRBSTwin-Strep-tagT7 terminatorpromoter for bacteriophage T7 RNA polymerase; T7 promoterlac operator5' proK leadertRNA-Pyl; Methanosarcina mazei tRNAPyl(TTA)3' proK trailertranscription terminator for bacteriophage T7 RNA polymerase; T7 terminatorpromoter for bacteriophage T7 RNA polymerase; T7 promoterlacI binding site5' proK leaderMethanocaldococcus jannaschii tRNApAzF(CTA)3' proK trailertranscription terminator for bacteriophage T7 RNA polymerase; T7 terminatorproK promotergRNA scaffoldproK terminatorproK promoterguide RNA scaffold for the CRISPR/Cas9 system; gRNA scaffoldproK terminatorEM7 promoterBleoRlambda t0 terminator

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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