LOCUS       Exported                2974 bp ds-DNA    circular SYN 03-11-2013
DEFINITION  Positive selection cloning vector with a lethal insert that allows 
            for efficient recovery of blunt-ended PCR products.
ACCESSION   EF694056
VERSION     .
KEYWORDS    pJET1.2
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2974)
  AUTHORS   Thermo Scientific (Fermentas)
  TITLE     Direct Submission
COMMENT     Linearize at Eco32I (EcoRV) to insert a blunt PCR product.
FEATURES             Location/Qualifiers
     source          1..2974
                     /organism="synthetic DNA construct"
                     /lab_host="Escherichia coli"
                     /mol_type="other DNA"
     CDS             complement(16..753)
                     /codon_start=1
                     /gene="eco47IR"
                     /product="mutant version of the Eco47I restriction 
                     endonuclease"
                     /function="
                     "
                     /note="Eco47I/T7"
                     /note="cleaves double-stranded DNA at GGWCC sites in the 
                     absence of protective methylation"
                     /protein_id="
                     "
                     /translation="MSKETSFVKNAEELAKQKMDAINPELSSKFKFLIKFLSQFPEACS
                     KPRSKKMQNKVGQEEHIEYLARSFHESRLPRKPTPPTTVPDEVVSIVLNISFNIQPENL
                     ERIKEEHRFSMAAENIVGDLLERYLAEKLEPSGRSGGRSPYSESYYAGWIWCSGTSVKA
                     VDFIHYDEKNNEWNLLQVKNRDNTENSSSSKIRDNTTIKKWFRTYSQRDATNWDNFPDE
                     VSSKNLNEEDFRAFVKNYLAKII"
     promoter        305..323
                     /note="T7 promoter"
                     /note="promoter for bacteriophage T7 RNA polymerase"
     misc_feature    328..422
                     /note="MCS"
                     /note="multiple cloning site with a central Eco32I (EcoRV) 
                     site"
     protein_bind    782..798
                     /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)."
     promoter        complement(806..836)
                     /note="lac UV5 promoter"
                     /note="E. coli lac promoter with an ""up"" mutation"
     rep_origin      complement(1163..1751)
                     /direction=LEFT
                     /note="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(1922..2782)
                     /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"
     promoter        complement(2783..2887)
                     /gene="bla"
                     /note="AmpR promoter"