LOCUS       Exported                6793 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Expression vector pL_delta_NG DNA, complete sequence.
ACCESSION   LC018348
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6793)
  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 6793)
  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 6793)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..6793
                     /organism="Expression vector pL_delta_NG"
                     /host="E. coli"
                     /mol_type="other DNA"
                     /label=lgfp_delta_1 and gfp
                     /note="lgfp_delta_1 and gfp"
                     /db_xref="taxon:1602145"
     CDS             complement(19..1134)
                     /codon_start=1
                     /gene="lacI"
                     /product="LacI"
                     /label=lacI
                     /protein_id="BAQ25583.1"
                     /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..2332
                     /codon_start=1
                     /gene="lgfp_delta_1"
                     /product="GFP deletion mutant"
                     /label=lgfp_delta_1
                     /protein_id="BAQ25584.1"
                     /translation="MSKGEELFTGVVPILVELDGDVNGQKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTFGYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFYKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKMEYNYNSHNVYIMADKPKNGIKV
                     NFKIRHNIKDGSVQL"
     gene            1796..2332
                     /gene="lgfp_delta_1"
                     /label=lgfp_delta_1
     terminator      2543..2629
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      2721..2748
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        2768..2859
                     /gene="bla"
                     /label=AmpR promoter
     CDS             2860..3720
                     /codon_start=1
                     /gene="Amp"
                     /product="beta-lactamase"
                     /label=Amp
                     /protein_id="BAQ25585.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPTAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     gene            2860..3720
                     /gene="Amp"
                     /label=Amp
     rep_origin      3743..4665
                     /label=ColE1-derived plasmid replication origin
                     /note="ColE1-derived plasmid replication origin"
     terminator      3750..3844
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     rep_origin      3932..4520
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(4741..5457)
                     /codon_start=1
                     /gene="gfpmut3"
                     /product="GFP"
                     /label=gfpmut3
                     /protein_id="BAQ25586.1"
                     /translation="MSKGEELFTGVVPILVELDGDVNGQKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTFGYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFYKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKMEYNYNSHNVYIMADKPKNGIKV
                     NFKIRHNIKDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMILLE
                     FVTAAGITHGMDELYK"
     gene            complement(4741..5457)
                     /gene="gfpmut3"
                     /label=gfpmut3
     promoter        complement(5482..5555)
                     /label=PLtetO-1 promoter
                     /note="modified phage lambda PL promoter with tet operator 
                     sites (Lutz and Bujard, 1997)"
     protein_bind    5512..5530
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    5537..5555
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     CDS             6107..6763
                     /codon_start=1
                     /gene="tetR"
                     /product="Tet R repressor"
                     /label=tetR
                     /protein_id="BAQ25587.1"
                     /translation="MVYAKDGRMDVMSRLDKSKVINSALELLNEVGIEGLTTRKLAQKL
                     GVEQPTLYWHVKNKRALLDALAIEMLDRHHTHFCPLEGESWQDFLRNNAKSFRCALLSH
                     RDGAKVHLGTRPTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEDQEHQ
                     VAKEERETPTTDSMPPLLRQAIELFDHQGAEPAFLFGLELIICGLEKQLKCESGS"
     gene            6107..6763
                     /gene="tetR"
                     /label=tetR
     CDS             6140..6763
                     /codon_start=1
                     /gene="tetR from transposon Tn10"
                     /product="tetracycline repressor TetR"
                     /label=TetR
                     /note="TetR binds to the tetracycline operator tetO to 
                     inhibit transcription. This inhibition can be relieved by 
                     adding tetracycline or doxycycline."
                     /translation="MSRLDKSKVINSALELLNEVGIEGLTTRKLAQKLGVEQPTLYWHV
                     KNKRALLDALAIEMLDRHHTHFCPLEGESWQDFLRNNAKSFRCALLSHRDGAKVHLGTR
                     PTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEDQEHQVAKEERETPTT
                     DSMPPLLRQAIELFDHQGAEPAFLFGLELIICGLEKQLKCESGS"