p2BBAD-phaB2-phaC2 vector (V015630)

LOCUS       62056_1930        8946 bp DNA     circular SYN 17-NOV-2021
DEFINITION  Expression vector p2BBAD-phaB2-phaC2, complete sequence.
ACCESSION   OL331254
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8946)
  AUTHORS   Tang R, Peng X, Weng C, Han Y.
  TITLE     The over-expression of phasin and regulator genes promoting the 
            synthesis of polyhydroxybutyrate in Cupriavidus necator H16 under 
            non-stress conditions
  JOURNAL   Appl Environ Microbiol, AEM0145821 (2021) In press
  PUBMED    34731058
REFERENCE   2  (bases 1 to 8946)
  AUTHORS   Tang R, Peng X, Weng C, Han Y.
  TITLE     Direct Submission
  JOURNAL   Submitted (31-OCT-2021) National Key Laboratory of Biochemical 
            Engineering, Institute of Process Engineering, Chinese Academy of 
            Sciences, 1 North 2nd Street, Zhongguancun PR, Beijing, Beijing 
            100190, China
REFERENCE   3  (bases 1 to 8946)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Appl 
            Environ Microbiol, AEM0145821 (2021) In press"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (31-OCT-2021) National Key Laboratory of Biochemical Engineering, 
            Institute of Process Engineering, Chinese Academy of Sciences, 1 
            North 2nd Street, Zhongguancun PR, Beijing, Beijing 100190, China"
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing 
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..8946
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      1023..1792
                     /label=pBBR1 oriV
                     /note="replication origin of the broad-host-range plasmid
                     pBBR1 from Bordetella bronchiseptica; requires the pBBR1 
                     Rep protein for replication"
     CDS             1793..2452
                     /label=pBBR1 Rep
                     /note="replication protein for the broad-host-range plasmid
                     pBBR1 from Bordetella bronchiseptica"
     primer_bind     3162..3178
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        3188..3206
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             complement(3248..4123)
                     /label=araC
                     /note="L-arabinose regulatory protein"
     promoter        4150..4434
                     /label=araBAD promoter
                     /note="promoter of the L-arabinose operon of E. coli; the
                     araC regulatory gene is transcribed in the opposite 
                     direction (Guzman et al., 1995)"
     regulatory      4492..4507
                     /regulatory_class="ribosome_binding_site"
     gene            4514..5260
                     /gene="phaB2"
                     /label=phaB2
     CDS             4514..5260
                     /codon_start=1
                     /transl_table=11
                     /gene="phaB2"
                     /product="PhaB2"
                     /EC_number="1.1.1.36"
                     /label=phaB2
                     /note="acetoacetyl-CoA reductase; derived from Cupriavidus 
                     necator H16"
                     /protein_id="UEP67409.1"
                     /translation="MAGQRIALVTGGMGGLGEAIAVRLLADGARVVVTHSVHNDHVAQW
                     LGTQRSAGREFTAFPVDVTDFASCQRCVSQVRSELGDVDILINNAGVTRDRTLRKMDKA
                     DWDFVLRTDLDSLFHMTRPLVEPMLARGWGRIVNISSVNASRGAFGQTNYAAAKAGVHG
                     FTKALALELARKGITVNTVSPGYLDTHMVTDMPAEILERDVLPTIPVGRLGKPAEVAAL
                     ISYLCSDDGAFVTGANFAINGGQHLQ"
     gene            5261..7036
                     /gene="phaC2"
                     /label=phaC2
     CDS             5261..7036
                     /codon_start=1
                     /transl_table=11
                     /gene="phaC2"
                     /product="PhaC2"
                     /label=phaC2
                     /note="poly(3-hydroxybutyrate) polymerase; derived from 
                     Cupriavidus necator H16"
                     /protein_id="UEP67410.1"
                     /translation="MRIDDRTSPPSLSAMPECAPQAAIPAQVWMDKQIRAGLARATGGL
                     SVISASLAGLDWALQLGISPAKIVQAWQVWLQHSIEDWPRLLPTAAEPPSVDLSDQRFA
                     DAGWQDWPYRCWRDAFMRNEAFWESLTQDIDGLSRHHQHIVAFCARQWLDLLAPGNVWW
                     MNPTIARTASSTFGANFLQGAHHWLQDASELLSKVPGLPAGRPALEFLPGRDVARTPGK
                     VVFRNALIELIEYAPALGANTVWREPVLIVPSWIMRYYILDLKPEDSLVRYLVESGHTV
                     FMISWKNPDASARDFGLDTYLEAGLLTALNTVHARCDGAHVHAAGYCLGGTLLATGAAM
                     LARDAAGGPLASMTLFASETDFHDPGELGLFIDKSSLATLDALMWSQGYLDGPQMKSAF
                     QMLNAQDLIWSRVMSEYLLGQRLRANDLVSWNRDTTRLPYRLHSECLHKLFLGNELATG
                     KLCVGGQPVALSDLDLPLFVVGTEHDHVSPWRSVYKLHLLTKAELTFLLTSGGHNAGIV
                     SEPGHPGRRYHVHTRLPAAPYLSPEQFLNVAVYRDGSWWPAWHDWLVAHSTARCTRQAP
                     GQGLADAPGTYVLEP"
     terminator      7047..7093
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     promoter        complement(7137..7155)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(7176..7192)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(7200..7216)
                     /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(7224..7254)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(7269..7290)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     CDS             complement(7698..8489)
                     /label=NeoR/KanR
                     /note="aminoglycoside phosphotransferase"