LOCUS       Exported                6410 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Expression vector pLGFPl DNA, complete sequence.
ACCESSION   LC018343
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6410)
  AUTHORS   Kobayashi H.
  TITLE     Inducible suppression of global translation by overuse of rare 
            codons
  JOURNAL   Appl. Environ. Microbiol. 81 (7), 2544-2553 (2015)
  PUBMED    25636849
REFERENCE   2  (bases 1 to 6410)
  AUTHORS   Kobayashi H.
  TITLE     Direct Submission
  JOURNAL   Submitted (07-JAN-2015) Contact:Hideki Kobayashi Agency for 
            Marine-Earth Science and Technology, Institute of Biogeosciences; 
            2-15 Natsushima, Yokosuka, Kanagawa 237-0061, Japan URL 
            :http://www.jamstec.go.jp/j/
REFERENCE   3  (bases 1 to 6410)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..6410
                     /organism="Expression vector pLGFPl"
                     /mol_type="other DNA"
                     /note="SC101 origin, host is E. coli, coding IPTG inducible
                     lgfp"
                     /db_xref="taxon:1602140"
     CDS             complement(19..1134)
                     /codon_start=1
                     /gene="lacI"
                     /product="LacI"
                     /label=lacI
                     /protein_id="BAQ25571.1"
                     /translation="MVYAKDGRMDVMKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKT
                     REKVEAAMAELNYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGAS
                     VVVSMVERSGVEACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSD
                     QTPINSIIFSHEDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQ
                     PIAEREGDWSAMSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISV
                     VGYDDTEDSSCYIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTT
                     LAPNTQTASPRALADSLMQLARQVSRLESGQ"
     gene            complement(19..1134)
                     /gene="lacI"
                     /label=lacI
     CDS             complement(19..1101)
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor"
                     /label=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)."
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     promoter        1723..1752
                     /label=trc promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    1760..1776
                     /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             1796..2512
                     /codon_start=1
                     /gene="lgfp"
                     /product="GFP"
                     /label=lgfp
                     /protein_id="BAQ25572.1"
                     /translation="MSKGEELFTGVVPILVELDGDVNGQKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTFGYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFYKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKMEYNYNSHNVYIMADKPKNGIKV
                     NFKIRHNIKDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMILLE
                     FVTAAGITHGMDELYK"
     gene            1796..2512
                     /gene="lgfp"
                     /label=lgfp
     terminator      2723..2809
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      2901..2928
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        2948..3039
                     /gene="bla"
                     /label=AmpR promoter
     CDS             3040..3900
                     /codon_start=1
                     /gene="Amp"
                     /product="beta-lactamase"
                     /label=Amp
                     /protein_id="BAQ25573.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPTAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     gene            3040..3900
                     /gene="Amp"
                     /label=Amp
     rep_origin      3923..6252
                     /label=SC101-derived plasmid replication origin
                     /note="SC101-derived plasmid replication origin"
     terminator      3930..4024
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     rep_origin      4516..4738
                     /label=pSC101 ori
                     /note="low-copy replication origin that requires the Rep101
                     protein"
     CDS             4786..5736
                     /codon_start=1
                     /gene="rep101"
                     /product="RepA protein needed for replication with the 
                     pSC101 origin"
                     /label=Rep101
                     /translation="MSELVVFKANELAISRYDLTEHETKLILCCVALLNPTIENPTRKE
                     RTVSFTYNQYAQMMNISRENAYGVLAKATRELMTRTVEIRNPLVKGFEIFQWTNYAKFS
                     SEKLELVFSEEILPYLFQLKKFIKYNLEHVKSFENKYSMRIYEWLLKELTQKKTHKANI
                     EISLDEFKFMLMLENNYHEFKRLNQWVLKPISKDLNTYSNMKLVVDKRGRPTDTLIFQV
                     ELDRQMDLVTELENNQIKMNGDKIPTTITSDSYLRNGLRKTLHDALTAKIQLTSFEAKF
                     LSDMQSKHDLNGSFSWLTQKQRTTLENILAKYGRI"
     terminator      6271..6357
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"