LOCUS       Exported                3653 bp ds-DNA    circular SYN 17-3-2013
DEFINITION  Vector for blue/white screening and for inducible bacterial 
            expression of tagged proteins.
ACCESSION   .
VERSION     .
KEYWORDS    pETBlue-2
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3653)
  AUTHORS   Novagen (EMD Millipore)
  TITLE     Direct Submission
COMMENT     This vector encodes a start codon that is positioned for optimal 
            expression.
FEATURES             Location/Qualifiers
     source          1..3653
                     /organism="synthetic DNA construct"
                     /lab_host="Escherichia coli"
                     /mol_type="other DNA"
     promoter        1..19
                     /note="T7 promoter"
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    20..44
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="lac operator"
                     /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             complement(54..491)
                     /codon_start=1
                     /gene="lacZ (truncated)"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /note="lacZ-alpha"
                     /translation="MTMITTPNLINISGGGGGARDPLGLQERVLAGLHVYTAVRPQASN
                     AYRWYRRRPAGAPVQNSDPRAPGISPWYISFLKLNKIILLANQDLSLAVVLQRRDWENP
                     GVTQLNRLAAHPPFASWRNSEEARTDRPSQQLRSLNGEWKL"
     RBS             265..270
                     /note="ribosome binding site"
     CDS             278..280
                     /codon_start=1
                     /product="start codon"
                     /note="ATG"
                     /translation="M"
     misc_feature    283..298
                     /note="blunt sites"
                     /note="blunt cutter sites that allow ORF insertion in all 
                     three reading frames"
     misc_feature    377..392
                     /note="blunt sites"
                     /note="blunt cutter sites that allow ORF insertion in all 
                     three reading frames"
     CDS             398..430
                     /codon_start=1
                     /product="HSV (herpes simplex virus) epitope tag"
                     /note="HSV tag"
                     /translation="QPELAPEDPED"
     CDS             437..454
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /note="6xHis"
                     /translation="HHHHHH"
     promoter        complement(541..569)
                     /note="tet promoter"
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     terminator      739..825
                     /gene="Escherichia coli rrnB"
                     /note="rrnB T1 terminator"
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      917..944
                     /note="rrnB T2 terminator"
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      1012..1059
                     /note="T7 terminator"
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      1096..1551
                     /direction=RIGHT
                     /note="f1 ori"
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     CDS             complement(1666..2526)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /note="AmpR"
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      2619..3207
                     /direction=RIGHT
                     /note="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     protein_bind    3606..3625
                     /map="
                     "
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="lac operator (symmetric)"
                     /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). The 
                     symmetric lac operator was optimized for tight binding of 
                     lac repressor."