pPPG2 vector (V003942)

Basic Vector Information

Vector Name:
pPPG2
Antibiotic Resistance:
Kanamycin
Length:
10157 bp
Type:
Expression vector
Replication origin:
p15A ori
Source/Author:
Kang MK, Eom JH, Kim Y, Um Y, Woo HM.
Promoter:
tac

pPPG2 vector Map

pPPG210157 bp50010001500200025003000350040004500500055006000650070007500800085009000950010000tac promoterlac operator(-)-alpha-pinene synthase, chloroplasticRBSgeranyl diphosphate synthaserrnB T1 terminatorrrnB T2 terminatorAmpR promoterKanRp15A ori

pPPG2 vector Sequence

LOCUS       V003942                10157 bp    DNA     circular SYN 18-DEC-2018
DEFINITION  Exported.
ACCESSION   V003942
VERSION     V003942
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 10157)
  AUTHORS   Kang MK, Eom JH, Kim Y, Um Y, Woo HM.
  TITLE     Biosynthesis of pinene from glucose using metabolically-engineered
            Corynebacterium glutamicum
  JOURNAL   Biotechnol. Lett. (2014) In press
   PUBMED   24930112
REFERENCE   2  (bases 1 to 10157)
  AUTHORS   Woo HM.
  TITLE     Direct Submission
  JOURNAL   Submitted (13-MAY-2014) Clean Energy Research Center, Korea
            Institute of Science and Technology, Hwarangno 14-gil 5,
            Seongbuk-gu, Seoul 136-791, Korea
REFERENCE   3  (bases 1 to 10157)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 10157)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing
            ##Assembly-Data-END##
            SGRef: number: 1; type: "Journal Article"; journalName: "Biotechnol.
            Lett. (2014) In press"
            SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
            (13-MAY-2014) Clean Energy Research Center, Korea Institute of
            Science and Technology, Hwarangno 14-gil 5, Seongbuk-gu, Seoul
            136-791, Korea"
            SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..10157
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        266..294
                     /label="tac promoter"
                     /note="strong E. coli promoter; hybrid between the trp and
                     lac UV5 promoters"
     protein_bind    302..318
                     /label="lac operator"
                     /bound_moiety="lac repressor encoded by lacI"
                     /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)."
     CDS             370..2256
                     /gene="PT1"
                     /label="(-)-alpha-pinene synthase, chloroplastic"
                     /note="(-)-alpha-pinene synthase, chloroplastic from Pinus
                     taeda. Accession#: Q84KL6"
     RBS             2266..2277
                     /note="strong bacterial ribosome binding site (Elowitz and
                     Leibler, 2000)"
     CDS             2284..3429
                     /codon_start=1
                     /product="geranyl diphosphate synthase"
                     /label="geranyl diphosphate synthase"
                     /note="AgGPPS2; codon-optimized for C. glutamicum"
                     /protein_id="AIF29191.1"
                     /translation="MAYSAMATMGYNGMAASCHTLHPTSPLKPFHGASTSLEAFNGEHM
                     GLLRGYSKRKLSSYKNPASRSSNATVAQLLNPPQKGKKAVEFDFNKYMDSKAMTVNEAL
                     NKAIPLRYPQKIYESMRYSLLAGGKRVRPVLCIAACELVGGTEELAIPTACAIEMIHTM
                     SLMHDDLPCIDNDDLRRGKPTNHKIFGEDTAVTAGNALHSYAFEHIAVSTSKTVGADRI
                     LRMVSELGRATGSEGVMGGQMVDIASEGDPSIDLQTLEWIHIHKTAMLLECSVVCGAII
                     GGASEIVIERARRYARCVGLLFQVVDDILDVTKSSDELGKTAGKDLISDKATYPKLMGL
                     EKAKEFSDELLNRAKGELSCFDPVKAAPLLGLADYVAFRQN"
     terminator      3656..3742
                     /label="rrnB T1 terminator"
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     terminator      3834..3861
                     /label="rrnB T2 terminator"
                     /note="transcription terminator T2 from the E. coli rrnB
                     gene"
     promoter        3880..3971
                     /label="AmpR promoter"
     CDS             complement(7919..8731)
                     /label="KanR"
                     /note="aminoglycoside phosphotransferase"
     rep_origin      complement(9326..9871)
                     /direction=LEFT
                     /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."

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