LOCUS       Exported                3257 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pGEM58ZNf(-), complete sequence.
ACCESSION   AF310245
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3257)
  AUTHORS   Hwang S-K.
  TITLE     Direct Submission
  JOURNAL   Submitted (02-OCT-2000) Div. of Biochemstry, NIAST, RDA, 249 
            Seodun-dong, Gwonsun-gu, Suwon 441-707, Korea
REFERENCE   2  (bases 1 to 3257)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..3257
                     /organism="Cloning vector pGEM58ZNf(-)"
                     /lab_host="Escherichia coli"
                     /mol_type="genomic DNA"
                     /db_xref="taxon:147091"
     misc_feature    5..121
                     /note="multicloning site; EcoRI, SacI, KpnI, NruI, NsiI, 
                     XbaI, EcoRV, BamHI, AvaI, SmaI, ApaI, SalI (AccI, HincII), 
                     PstI, NotI, SpeI, EcoRV, SacII, NcoI, SphI, HindIII; 
                     EcoRI-HindIII fragment of pUC58 (AF253496) was ligated with
                     the EcoRI-HindIII gap of pGEM-3Zf(-); insertion did not 
                     affect a blue/white selection"
     regulatory      95..104
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     regulatory      127..146
                     /regulatory_class="promoter"
                     /note="SP6"
     promoter        complement(128..146)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(164..180)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    188..204
                     /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(212..242)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    257..278
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      complement(566..1154)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(1325..2185)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /function="ampicillin resistance"
                     /label=bla
                     /protein_id="AAG48609.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     gene            complement(1325..2185)
                     /gene="bla"
                     /label=bla
     promoter        complement(2186..2290)
                     /gene="bla"
                     /label=AmpR promoter
     misc_feature    2622..3077
                     /label=phage f1 (-) region
                     /note="phage f1 (-) region"
     rep_origin      2622..3077
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     CDS             complement(3069..168)
                     /codon_start=1
                     /gene="lacZ"
                     /product="beta-galactosidase"
                     /label=lacZ
                     /note="synthetic"
                     /protein_id="AAG48608.1"
                     /translation="MTMITPSYLGDTIEYSSLHAPGRHGRGISLVRPPAGRRARDPISR
                     CIREVPSSNSPYSESYYNSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEARTDR
                     PSQQLRSLNGEWKL"
     gene            complement(3069..168)
                     /gene="lacZ"
                     /label=lacZ
     primer_bind     3218..3234
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     regulatory      3241..3
                     /regulatory_class="promoter"
                     /note="T7"
     promoter        3241..2
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"