pBla_T7 vector (V016259)

Basic Vector Information

Vector Name:
pBla_T7
Antibiotic Resistance:
Bleomycin
Length:
2593 bp
Type:
Cloning vector
Replication origin:
p15A ori
Source/Author:
Hashimoto K, Fischer EC, Romesberg FE.

pBla_T7 vector Map

pBla_T72593 bp600120018002400p15A oriT7 promoterRBSblaT7 terminatorpromoter for bacteriophage T7 RNA polymerase; T7 promoter5' leader from serTtRNA Ser(UGA)3' UTR from serTtranscription terminator for bacteriophage T7 RNA polymerase; T7 terminatorEM7 promoterBleoRlambda t0 terminator

pBla_T7 vector Sequence

LOCUS       62056_4090        2593 bp DNA     circular SYN 16-JUN-2021
DEFINITION  Cloning vector pBla_T7, complete sequence.
ACCESSION   MW816821
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2593)
  AUTHORS   Hashimoto K, Fischer EC, Romesberg FE.
  TITLE     Efforts toward Further Integration of an Unnatural Base Pair into 
            the Biology of a Semisynthetic Organism
  JOURNAL   J Am Chem Soc (2021) In press
  PUBMED    34096294
REFERENCE   2  (bases 1 to 2593)
  AUTHORS   Hashimoto K.
  TITLE     Direct Submission
  JOURNAL   Submitted (25-MAR-2021) Department of Chemistry, The Scripps 
            Research Institute, 10550 North Torrey Pines Road, La Jolla, CA 
            92037, USA
REFERENCE   3  (bases 1 to 2593)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "J Am Chem 
            Soc (2021) In press"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (25-MAR-2021) Department of Chemistry, The Scripps Research 
            Institute, 10550 North Torrey Pines Road, La Jolla, CA 92037, USA"
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing 
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..2593
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      109..654
                     /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        807..825
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             840..862
                     /label=RBS
                     /note="efficient ribosome binding site from bacteriophage
                     T7 gene 10 (Olins and Rangwala, 1989)"
     gene            871..1701
                     /gene="bla"
                     /label=bla
     CDS             join(871..1048,1067..1701)
                     /codon_start=1
                     /transl_table=11
                     /gene="bla"
                     /product="beta-lactamase TEM-1"
                     /label=bla
                     /protein_id="QWO78777.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLDSGKILESFRRAVLSRIDAGQEQLGRRIHYSQNDLVEYSPVTEKHLTDGMTVREL
                     CSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRWEPELNEAIPNDERDTT
                     MPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSALPAGWFIADKSGAGER
                     GSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGASLIKHW"
     misc_feature    1044..1048
                     /gene="bla"
                     /label=UBP
                     /note="UBP"
     misc_feature    1049..1054
                     /gene="bla"
                     /label=BsaI
                     /note="BsaI"
     misc_feature    1055..1060
                     /gene="bla"
                     /label=KpnI
                     /note="KpnI"
     misc_feature    1061..1066
                     /gene="bla"
                     /label=BsaI
                     /note="BsaI"
     misc_feature    1067..1073
                     /gene="bla"
                     /label=UBP
                     /note="UBP"
     terminator      1733..1780
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     regulatory      1810..1828
                     /note="promoter for bacteriophage T7 RNA polymerase; T7 
                     promoter"
                     /regulatory_class="promoter"
     misc_feature    1840..1846
                     /label=5' leader from serT
                     /note="5' leader from serT"
     misc_feature    1847..1908
                     /label=tRNA Ser(UGA)
                     /note="tRNA Ser(UGA)"
     misc_feature    1861..1866
                     /label=BsaI
                     /note="BsaI"
     misc_feature    1867..1872
                     /label=KpnI
                     /note="KpnI"
     misc_feature    1873..1878
                     /label=BsaI
                     /note="BsaI"
     misc_feature    1909..1951
                     /label=3' UTR from serT
                     /note="3' UTR from serT"
     regulatory      1952..1999
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase; T7 terminator"
                     /regulatory_class="terminator"
     promoter        2047..2094
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter"
     CDS             2113..2484
                     /label=BleoR
                     /note="antibiotic-binding protein"
     terminator      2499..2593
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"

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