LOCUS       Exported                4779 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pLACT7, complete sequence.
ACCESSION   U16722
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4779)
  AUTHORS   Chong S, Garcia GA.
  TITLE     A versatile and general prokaryotic expression vector, pLACT7
  JOURNAL   BioTechniques 17 (4), 686-681 (1994)
  PUBMED    7833029
REFERENCE   2  (bases 1 to 4779)
  AUTHORS   Garcia GA.
  TITLE     Direct Submission
  JOURNAL   Submitted (28-OCT-1994) George A. Garcia, College of Pharmacy, 
            University of Michigan, 428 Church St., Ann Arbor, MI 48109-1065
REFERENCE   3  (bases 1 to 4779)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..4779
                     /organism="Cloning vector pLACT7"
                     /mol_type="genomic DNA"
                     /db_xref="taxon:37153"
     primer_bind     149..165
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     regulatory      170..220
                     /regulatory_class="terminator"
                     /note="from T7"
     terminator      174..220
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     misc_feature    221..289
                     /label=multiple cloning site
                     /note="multiple cloning site"
     CDS             complement(254..286)
                     /codon_start=1
                     /product="leader peptide from bacteriophage T7 gene 10"
                     /label=T7 tag (gene 10 leader)
                     /note="promotes efficient translation in E. coli"
                     /translation="MASMTGGQQMG"
     misc_feature    284
                     /label=NdeI site
                     /note="NdeI site"
     regulatory      295..303
                     /regulatory_class="ribosome_binding_site"
     RBS             295..303
                     /label=Shine-Dalgarno sequence
                     /note="full consensus sequence for ribosome-binding sites 
                     upstream of start codons in E. coli; complementary to a 
                     region in the 3 end of the 16S rRNA (Chen et al., 1994)"
     promoter        complement(310..328)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      312..328
                     /regulatory_class="promoter"
                     /note="from T7"
     primer_bind     complement(342..358)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     regulatory      362..386
                     /regulatory_class="promoter"
                     /note="for lac gene"
     protein_bind    366..382
                     /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)."
     promoter        complement(390..420)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    435..456
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        981..1058
                     /gene="lacI"
                     /label=lacI promoter
     CDS             1059..2141
                     /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"
     rep_origin      complement(2580..3168)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(3339..4199)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(4200..4304)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(4331..7)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"