LOCUS       Exported                7876 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pJLRCS, complete sequence.
ACCESSION   KX273378
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7876)
  AUTHORS   Trachtmann N, Alvarez Fong KF, Guitart Font E, Sprenger GA.
  TITLE     Construction of chromosomally encoded lacZ and gfp reporter strains 
            of Escherichia coli for global regulation of metabolism
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 7876)
  AUTHORS   Trachtmann N, Alvarez Fong KF, Guitart Font E, Sprenger GA.
  TITLE     Direct Submission
  JOURNAL   Submitted (24-MAY-2016) Institute of microbiology, Stuttgart 
            university, Allmandring 31, Stuttgart 70569, Germany
REFERENCE   3  (bases 1 to 7876)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing 
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..7876
                     /organism="Cloning vector pJLRCS"
                     /mol_type="other DNA"
                     /label=derived from Escherichia coli Lj110
                     /note="derived from Escherichia coli Lj110"
                     /db_xref="taxon:1941121"
     misc_feature    6..303
                     /label=flanking arm for integration
                     /note="flanking arm for integration"
     regulatory      400..520
                     /regulatory_class="terminator"
                     /note="5S terminator"
     terminator      522..608
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     regulatory      523..725
                     /regulatory_class="terminator"
                     /note="rrnB T1T2 terminator"
     terminator      700..727
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        746..837
                     /gene="bla"
                     /label=AmpR promoter
     CDS             838..1698
                     /codon_start=1
                     /gene="bla"
                     /product="beta lactamase"
                     /function="ampicillin resistance"
                     /label=bla
                     /protein_id="APZ86790.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     gene            838..1698
                     /gene="bla"
                     /label=bla
     rep_origin      1860..2459
                     /label=pMB1 origin of replication
                     /note="pMB1 origin of replication"
     rep_origin      1869..2457
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     misc_feature    2643..2783
                     /label=bom
                     /note="basis of mobility region from pBR322"
     CDS             complement(2885..3076)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(4094..5176)
                     /codon_start=1
                     /gene="lacI"
                     /product="Lac repressor"
                     /label=lacI
                     /protein_id="APZ86791.1"
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     gene            complement(4094..5176)
                     /gene="lacI"
                     /label=lacI
     promoter        complement(5177..5254)
                     /gene="lacI (mutant)"
                     /label=lacIq promoter
                     /note="In the lacIq allele, a single base change in the 
                     promoter boosts expression of the lacI gene about 10-fold."
     misc_feature    5358..5881
                     /label=flanking arm for integration region
                     /note="flanking arm for integration region"
     protein_bind    complement(5912..5959)
                     /label=FRT
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="FLP-mediated recombination occurs in the 8-bp core 
                     sequence TCTAGAAA (Turan and Bode, 2011)."
     CDS             complement(5995..6633)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /function="chloramphenicol resistance"
                     /label=cat
                     /protein_id="APZ86792.1"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPELRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRCLMNTTVLR"
     gene            complement(5995..6633)
                     /gene="cat"
                     /label=cat
     promoter        complement(6634..6736)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     protein_bind    complement(6842..6875)
                     /label=FRT (minimal)
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="supports FLP-mediated excision but not integration 
                     (Turan and Bode, 2011)"
     regulatory      7066..7094
                     /regulatory_class="promoter"
                     /note="tac promoter"
     promoter        7067..7095
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    7103..7119
                     /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             7159..7875
                     /codon_start=1
                     /gene="gfp"
                     /product="green fluorescence protein"
                     /label=gfp
                     /protein_id="APZ86793.1"
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTFAYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     gene            7159..7875
                     /gene="gfp"
                     /label=gfp