LOCUS       Exported                5071 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pKSM710, complete sequence.
ACCESSION   U04893
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5071)
  AUTHORS   Maneewannakul S, Maneewannakul K, Ippen-Ihler K.
  TITLE     The pKSM710 vector cassette provides tightly regulated lac and T7lac
            promoters and strategies for manipulating N-terminal protein 
            sequences
  JOURNAL   Plasmid 31 (3), 300-307 (1994)
  PUBMED    8058823
REFERENCE   2  (bases 1 to 5071)
  AUTHORS   Maneewannakul S.
  TITLE     Direct Submission
  JOURNAL   Submitted (06-JAN-1994) Sumit Maneewannakul, Cellular and 
            Developmental Biology, Harvard University, 16 Divinity Avenue, 
            Cambridge, MA 02138, USA
REFERENCE   3  (bases 1 to 5071)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..5071
                     /organism="Cloning vector pKSM710"
                     /lab_host="Escherichia coli"
                     /mol_type="genomic DNA"
                     /db_xref="taxon:31861"
     regulatory      24..70
                     /regulatory_class="terminator"
     terminator      27..75
                     /label=fd terminator
                     /note="central terminator from bacteriophage fd (Otsuka and
                     Kunisawa, 1982)"
     regulatory      134..180
                     /regulatory_class="terminator"
     terminator      137..185
                     /label=fd terminator
                     /note="central terminator from bacteriophage fd (Otsuka and
                     Kunisawa, 1982)"
     rep_origin      313..768
     rep_origin      complement(921..1515)
                     /direction=LEFT
     rep_origin      complement(927..1515)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(1686..2546)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /function="antibiotic resistance"
                     /label=bla
                     /protein_id="AAA61618.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     gene            complement(1686..2546)
                     /gene="bla"
                     /label=bla
     regulatory      complement(2720..2774)
                     /regulatory_class="terminator"
     CDS             complement(2861..3520)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyl methylase"
                     /function="chloramphenicol resistance"
                     /label=cat
                     /protein_id="AAA61619.1"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     gene            complement(2861..3520)
                     /gene="cat"
                     /label=cat
     promoter        complement(3521..3623)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     regulatory      3881..3910
                     /regulatory_class="promoter"
                     /note="lambda Pl promoter"
     promoter        4109..4127
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      4109..4125
                     /regulatory_class="promoter"
                     /note="T7 promoter"
     protein_bind    4128..4152
                     /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        4286..4316
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     regulatory      4286..4315
                     /regulatory_class="promoter"
                     /note="lacUV5 promoter"
     protein_bind    4324..4340
                     /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     complement(4486..4502)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     terminator      4693..4740
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     regulatory      4698..4739
                     /regulatory_class="terminator"
                     /note="T7 terminator; T-phi"
     terminator      4793..4879
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     regulatory      4796..4839
                     /regulatory_class="terminator"
                     /note="rrnBT1 terminator"
     regulatory      4970..4999
                     /regulatory_class="terminator"
                     /note="rrnBT2 terminator"
     terminator      4971..4998
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"