LOCUS       Exported                6267 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Expression vector pSK9, complete sequence.
ACCESSION   JQ322997
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6267)
  AUTHORS   Kuchmina E, Wallner T, Kryazhov S, Zinchenko VV, Wilde A.
  TITLE     An expression system for regulated protein production in 
            Synechocystis sp. PCC 6803 and its application for construction of a
            conditional knockout of the ferrochelatase enzyme
  JOURNAL   J. Biotechnol. 162 (1), 75-80 (2012)
  PUBMED    22749908
REFERENCE   2  (bases 1 to 6267)
  AUTHORS   Zinchenko VV, Kryazhov SV.
  TITLE     Direct Submission
  JOURNAL   Submitted (27-DEC-2011) Dept. of Genetics, Biology Division, Moscow 
            State University, Moscow 119899, Russia
REFERENCE   3  (bases 1 to 6267)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..6267
                     /organism="Expression vector pSK9"
                     /mol_type="other DNA"
                     /db_xref="taxon:1243990"
     misc_feature    join(4..621,626..655)
                     /note="derived from cloning vector pBluescript II KS (+) in
                     INSD: X52327"
     rep_origin      complement(6..461)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     603..619
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        626..644
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     misc_feature    656..1145
                     /note="p6803-platform; derived from Synechocystis sp. PCC 
                     6803 chromosome in INSD: BA000022"
     misc_feature    1146..2580
                     /note="derived from cloning vector pACYC184 in INSD: 
                     X06403"
     CDS             complement(1816..2475)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=cat
                     /protein_id="AFV26885.1"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     gene            complement(1816..2475)
                     /gene="cat"
                     /label=cat
     misc_feature    2581..3058
                     /note="leader region of petJ gene derived from 
                     Synechocystis sp. PCC 6803 chromosome in INSD: BA000022"
     misc_feature    3059..4063
                     /note="p6803-platform; derived from Synechocystis sp. PCC 
                     6803 chromosome in INSD: BA000022"
     misc_feature    4064..6267
                     /note="derived from cloning vector pBluescript II KS (+) in
                     INSD: X52327"
     promoter        complement(4082..4100)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(4121..4137)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    4145..4161
                     /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(4169..4199)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4214..4235
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      complement(4523..5111)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(5282..6142)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=bla
                     /protein_id="AFV26886.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     gene            complement(5282..6142)
                     /gene="bla"
                     /label=bla
     promoter        complement(6143..6247)
                     /gene="bla"
                     /label=AmpR promoter