LOCUS       Exported                4451 bp ds-DNA     circular SYN 17-DEC-2018
DEFINITION  Expression vector pEamTA, complete sequence.
ACCESSION   EF565824
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4451)
  AUTHORS   Reisinger C, Kern A, Fesko K, Schwab H.
  TITLE     An efficient plasmid vector for expression cloning of large numbers 
            of PCR fragments in Escherichia coli
  JOURNAL   Appl. Microbiol. Biotechnol. 77 (1), 241-244 (2007)
  PUBMED    17786428
REFERENCE   2  (bases 1 to 4451)
  AUTHORS   Reisinger C, Kern A, Fesko K, Schwab H.
  TITLE     Direct Submission
  JOURNAL   Submitted (19-APR-2007) 1 Research Centre Applied Biocatalysis, 
            Petersgasse 14/4, Graz, Styria A-8010, Austria
REFERENCE   3  (bases 1 to 4451)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..4451
                     /organism="Expression vector pEamTA"
                     /mol_type="other DNA"
                     /label=derived from pMS470delta8 expression vector
                     /note="derived from pMS470delta8 expression vector"
                     /db_xref="taxon:443865"
     RBS             40..62
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     regulatory      55..61
                     /regulatory_class="ribosome_binding_site"
     misc_feature    69
                     /label=single 3-T overhang in linearized vector
                     /note="single 3-T overhang in linearized vector"
     misc_feature    517
                     /label=single 3-T overhang in linearized vector
                     /note="single 3-T overhang in linearized vector"
     regulatory      549..949
                     /regulatory_class="terminator"
                     /note="rrnB terminator"
     terminator      746..832
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      924..951
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        971..1062
                     /gene="bla"
                     /label=AmpR promoter
     CDS             1063..1923
                     /codon_start=1
                     /gene="bla"
                     /product="beta lactamase"
                     /label=bla
                     /note="ampicillin resistance"
                     /protein_id="ABQ45973.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     gene            1063..1923
                     /gene="bla"
                     /label=bla
     misc_feature    1851..1854
                     /gene="bla"
                     /label=mutation destroys Eam1105I restriction enzyme site
                     /note="mutation destroys Eam1105I restriction enzyme site"
     rep_origin      2068..2741
                     /label=sequence derived from pUC ori
                     /note="sequence derived from pUC ori"
     rep_origin      2094..2682
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(2993..4084)
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor protein"
                     /label=lacI
                     /protein_id="ABQ45974.1"
                     /translation="MVNVKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAM
                     AELNYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVER
                     SGVEACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSII
                     FSHEDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGD
                     WSAMSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTED
                     SSCYIPPLTTSKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTA
                     SPRALADSLMQLARQVSRLESGQ"
     gene            complement(2993..4084)
                     /gene="lacI"
                     /label=lacI
     CDS             complement(2993..4075)
                     /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
                     YIPPLTTSKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     promoter        complement(4076..4153)
                     /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."
     regulatory      4238..4444
                     /regulatory_class="promoter"
                     /gene="tac"
     gene            4238..4444
                     /gene="tac"
                     /label=tac
     promoter        4383..4411
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     regulatory      4383..4388
                     /regulatory_class="minus_35_signal"
                     /gene="tac"
     regulatory      4405..4410
                     /regulatory_class="minus_10_signal"
                     /gene="tac"
     protein_bind    4419..4435
                     /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)."