LOCUS       Exported                3455 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  PCR cloning vector pAnti-Kit-Barnase, complete sequence.
ACCESSION   DQ839632
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3455)
  AUTHORS   Krotkova A.
  TITLE     Direct Submission
  JOURNAL   Submitted (09-JUL-2006) Biosave Ltd./GmbH, Bol. Cheremushkinskaya 
            11/1, Moscow 113447, Russia
REFERENCE   2  (bases 1 to 3455)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..3455
                     /organism="PCR cloning vector pAnti-Kit-Barnase"
                     /mol_type="other DNA"
                     /db_xref="taxon:398030"
     promoter        24..128
                     /gene="bla"
                     /label=AmpR promoter
     CDS             129..989
                     /codon_start=1
                     /product="beta-lactamase"
                     /label=beta-lactamase
                     /protein_id="ABI13484.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      1138..1803
                     /label=pUC
                     /note="pUC"
     rep_origin      1160..1748
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     regulatory      2024..2145
                     /regulatory_class="promoter"
                     /note="lac"
     protein_bind    2036..2057
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        2072..2102
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2110..2126
                     /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)."
     primer_bind     2134..2150
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             2146..2742
                     /codon_start=1
                     /product="LacZ/MRS/barnase fusion protein"
                     /label=LacZ/MRS/barnase fusion protein
                     /protein_id="ABI13485.1"
                     /translation="MTMITPSAQLTLTKGNKSWVPGPPSRSTVSISLRIPILVWHFSFT
                     AEKEKCHTSIGYLEFLQPGGSTSSRGGRASIVSPKWGPPPVELAQVINTFDGVADYLQT
                     YHKLPDNYITKSEAQALGWVASKGNLADVAPGKSIGGDIFSNREGKLPGKSGRTWREAD
                     INYTSGFRNSDRILYSSDWLIYKTTDHYQTFTKIR"
     promoter        2171..2189
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     misc_structure  2207..2359
                     /note="MRS; restriction site polylinker"
     primer_bind     2219..2235
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     stem_loop       2259..2306
                     /label=XcmI-XcmI hairpin sequence
                     /note="XcmI-XcmI hairpin sequence"
     primer_bind     complement(2339..2355)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        complement(2368..2386)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     CDS             2410..2742
                     /codon_start=1
                     /product="ribonuclease from Bacillus amyloliquefaciens"
                     /label=barnase
                     /note="The barnase gene is lethal in standard bacterial 
                     transformation strains."
                     /translation="AQVINTFDGVADYLQTYHKLPDNYITKSEAQALGWVASKGNLADV
                     APGKSIGGDIFSNREGKLPGKSGRTWREADINYTSGFRNSDRILYSSDWLIYKTTDHYQ
                     TFTKIR"
     promoter        complement(2812..2830)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2840..2856)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     rep_origin      2998..3453
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     rep_origin      3016..3322
                     /label=f1(+)
                     /note="f1(+)"