LOCUS       Exported                3197 bp ds-DNA    circular SYN 24-6-2015
DEFINITION  Vector for standard cloning, efficient in vitro RNA synthesis, and 
            ssDNA production. Identical to pGEM(R)?3Zf(+) except for the 
            orientation of the f1 origin.
ACCESSION   X65307
VERSION     .
KEYWORDS    pGEM-3Zf(-)
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3197)
  AUTHORS   Promega
  TITLE     Direct Submission
COMMENT     
FEATURES             Location/Qualifiers
     source          1..3197
                     /organism="synthetic DNA construct"
                     /lab_host="Escherichia coli"
                     /mol_type="other DNA"
     misc_feature    5..61
                     /gene="
                     "
                     /note="MCS"
                     /note="pUC18/19 multiple cloning site"
     promoter        complement(68..86)
                     /note="SP6 promoter"
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(104..120)
                     /note="M13 rev"
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    128..144
                     /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(152..182)
                     /note="lac promoter"
                     /note="promoter for the E. coli lac operon"
     rep_origin      complement(506..1094)
                     /direction=LEFT
                     /note="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(1265..2125)
                     /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(2126..2230)
                     /gene="bla"
                     /note="AmpR promoter"
     rep_origin      2562..3017
                     /direction=RIGHT
                     /note="f1 ori"
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     CDS             complement(3009..108)
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /note="lacZ-alpha"
                     /translation="MTMITPSYLGDTIEYSSLHACRSTLEDPRVPSSNSPYSESYYNSL
                     AVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEARTDRPSQQLRSLNGEWKL"
     primer_bind     3158..3174
                     /note="M13 fwd"
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        3181..2
                     /note="T7 promoter"
                     /note="promoter for bacteriophage T7 RNA polymerase"