LOCUS       Exported                4825 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pQTEV2, complete sequence.
ACCESSION   EF429248
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4825)
  AUTHORS   Ergin A, Thiel A, Duchmann R, Sieper J, Bussow K, Adam T.
  TITLE     Homologous High-Throughput Expression and Purification of Highly 
            Conserved E. coli Proteins
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 4825)
  AUTHORS   Ergin A, Adam T, Bussow K.
  TITLE     Direct Submission
  JOURNAL   Submitted (09-FEB-2007) Gastroenterology, Charite Berlin (CBF), 
            Hindenburgdamm 30, Berlin 12200, Germany
REFERENCE   3  (bases 1 to 4825)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..4825
                     /organism="Cloning vector pQTEV2"
                     /mol_type="other DNA"
                     /db_xref="taxon:429152"
     promoter        10..54
                     /label=T5 promoter
                     /note="bacteriophage T5 promoter for E. coli RNA 
                     polymerase, with embedded lac operator"
     protein_bind    30..46
                     /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)."
     regulatory      30..35
                     /regulatory_class="minus_35_signal"
     regulatory      48..53
                     /regulatory_class="minus_10_signal"
     protein_bind    62..78
                     /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)."
     RBS             97..108
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     regulatory      101..107
                     /regulatory_class="ribosome_binding_site"
     misc_feature    115
                     /note="translation start site for inserts cloned into the 
                     multiple cloning site"
     CDS             121..141
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=7xHis
                     /translation="HHHHHHH"
     protein_bind    154..178
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     misc_recomb     154..177
                     /label=attB1 recombination site
                     /note="attB1 recombination site"
     misc_feature    178..198
                     /note="Tobacco etch virus (TEV) protease 
                     recognition/cleavage region"
     CDS             178..198
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     misc_feature    195..230
                     /note="multiple cloning site; includes BamHI, SphI, NotI 
                     restriction sites"
     misc_recomb     235..257
                     /label=attB2 recombination site
                     /note="attB2 recombination site"
     protein_bind    complement(236..260)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     regulatory      282..376
                     /regulatory_class="terminator"
                     /note="lambda t0 transcriptional terminator"
     terminator      282..376
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             420..1079
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     regulatory      1138..1235
                     /regulatory_class="terminator"
                     /note="rrnB1 T1 transcriptional terminator"
     terminator      1144..1230
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             complement(1323..2405)
                     /codon_start=1
                     /gene="lacI"
                     /allele="Q"
                     /product="LacI"
                     /label=lacI
                     /note="lac repressor; overproducing allele; similar to lac 
                     repressor encoded by GenBank Accession Number U78872"
                     /protein_id="ABO25745.1"
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     gene            complement(1323..2405)
                     /gene="lacI"
                     /allele="Q"
                     /label=lacI
     promoter        complement(2406..2483)
                     /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    2675..2815
                     /label=bom
                     /note="basis of mobility region from pBR322"
     rep_origin      complement(3001..3589)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(3760..4620)
                     /codon_start=1
                     /gene="bla"
                     /product="beta lactamase precursor"
                     /label=bla
                     /note="penicillinase; similar to the protein encoded by 
                     GenBank Accession Number AF074376"
                     /protein_id="ABO25746.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     gene            complement(3760..4620)
                     /gene="bla"
                     /label=bla
     promoter        complement(4621..4725)
                     /gene="bla"
                     /label=AmpR promoter