pDS261 vector (V016375)

LOCUS       62056_8595        6001 bp DNA     circular SYN 27-APR-2021
DEFINITION  Suicide vector pDS261, complete sequence.
ACCESSION   MW429486
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6001)
  AUTHORS   Susanti D, Frazier MC, Mukhopadhyay B.
  TITLE     A Genetic System for Methanocaldococcus jannaschii: An Evolutionary 
            Deeply Rooted Hyperthermophilic Methanarchaeon
  JOURNAL   Front Microbiol 10, 1256 (2019)
  PUBMED    31333590
REFERENCE   2  (bases 1 to 6001)
  AUTHORS   Khairunisa BH, Susanti D, Mukhopadhyay B.
  TITLE     Direct Submission
  JOURNAL   Submitted (03-JAN-2021) Biochemistry, Virginia Tech, 340 West Campus
            Drive, Blacksburg, VA 24061, USA
REFERENCE   3  (bases 1 to 6001)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Front 
            Microbiol 10, 1256 (2019)"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (03-JAN-2021) Biochemistry, Virginia Tech, 340 West Campus Drive, 
            Blacksburg, VA 24061, USA"
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing 
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..6001
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(3..458)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     primer_bind     600..616
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        626..644
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     670..686
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             698..1000
                     /codon_start=1
                     /transl_table=11
                     /product="hypothetical protein"
                     /label=hypothetical protein
                     /note="part of mj_0749; upstream of fprA (mj_0748)"
                     /protein_id="QUC68721.1"
                     /translation="AYLLLALFLVLGFIYPMFFCRYVCPVGSLLSIFARFSIFKLKLDE
                     NKCVGCRLCERKCPMQIKITEKIDQMECIRCFECMSVCKKGALSFSAFTKNTKKE"
     misc_feature    1001..1212
                     /label=upstream of fprA (mj_0748)
                     /note="upstream of fprA (mj_0748)"
     regulatory      1213..1528
                     /note="Methanocaldococcus FS.406-22 S-layer promoter (Psla,
                     mfs40622_1341)"
                     /regulatory_class="promoter"
     gene            1529..2746
                     /gene="hmgA"
                     /locus_tag="mj_0705"
                     /label=hmgA
     CDS             1529..2746
                     /codon_start=1
                     /transl_table=11
                     /gene="hmgA"
                     /locus_tag="mj_0705"
                     /product="3-hydroxy-3-methylglutaryl coenzyme A reductase"
                     /label=hmgA
                     /protein_id="QUC68722.1"
                     /translation="MENYNDILEKMLNGEIKPYQLDKMFGSKIATEIRRKFIEKKVGIE
                     FKHICNYSIDEEMAMKKNIENMIGAIQIPLGFAGPLKINGEYAKGEFYIPLATTEGALV
                     ASVNRGCSIITKCGGATVRVIDDKMTRAPCLKTKSVVDAIKVRDWIRENFERIKEVAES
                     TTRHGKLIKIEPILIVGRNLYPRFVFKTGDAMGMNMVTIATEKACNFIEGELKKEGIFV
                     KTVAVSGNACVDKKPSGMNLINGRGKSIVAEVFLTEKEVNKYLKTTSQAIAEVNRLKNY
                     IGSAISNSMGFNAHYANIIGAIFLATGQDEAHIVEGSLGITMAEVEDDGLYFSVTLPDV
                     PIGTVGGGTRVETQKECLEMLGCYGDNKALKFAEIVGAAVLAGELSLLGALAAGHLGKA
                     HQELGR"
     regulatory      2747..2780
                     /note="S-layer terminator from Methanocaldococcus FS.406-22
                     (Tsla, mfs40622_1341)"
                     /regulatory_class="terminator"
     regulatory      2781..3056
                     /note="flagellin B1B2 (mj_0891-0892) operon promoter of 
                     Methanocaldococcus jannaschii (PflaB1B2)"
                     /regulatory_class="promoter"
     CDS             3066..3131
                     /label=3xFLAG
                     /note="three tandem FLAG(R) epitope tags, followed by an 
                     enterokinase cleavage site"
     CDS             3147..3170
                     /label=Strep-Tag II
                     /note="peptide that binds Strep-Tactin(R), an engineered 
                     form
                     of streptavidin"
     CDS             3207..3230
                     /label=Strep-Tag II
                     /note="peptide that binds Strep-Tactin(R), an engineered 
                     form
                     of streptavidin"
     primer_bind     complement(3760..3776)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        complement(3813..3831)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(3852..3868)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(3876..3892)
                     /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)."
     promoter        complement(3900..3930)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(3945..3966)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(4254..4842)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(5016..5873)
                     /label=AmpR
                     /note="beta-lactamase"
     promoter        complement(5874..5978)
                     /label=AmpR promoter