LOCUS       40924_27024        5041 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pKSM713, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5041)
  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 5041)
  AUTHORS   Maneewannakul S.
  TITLE     Direct Submission
  JOURNAL   Submitted (06-JAN-1994) Sumit Maneewannakul, Harvard University, 
            Cellular and Developmental Biology, 16 Divinity Avenue, Cambridge, 
            MA 02138, USA
REFERENCE   3  (bases 1 to 5041)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 5041)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Plasmid"; 
            date: "1994"; volume: "31"; issue: "3"; pages: "300-307"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (06-JAN-1994) Sumit Maneewannakul, Harvard University, Cellular and 
            Developmental Biology, 16 Divinity Avenue, Cambridge, MA 02138, USA"
COMMENT     SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..5041
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        20..38
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    39..63
                     /label=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        197..227
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an 'up' mutation"
     protein_bind    235..251
                     /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(397..413)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     terminator      604..651
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      704..790
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     terminator      882..909
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB
                     gene"
     terminator      1009..1057
                     /label=fd terminator
                     /note="central terminator from bacteriophage fd (Otsuka and
                     Kunisawa, 1982)"
     regulatory      1116..1162
                     /label=bacteriophage fd terminator
                     /note="bacteriophage fd terminator"
                     /regulatory_class="terminator"
     terminator      1119..1167
                     /label=fd terminator
                     /note="central terminator from bacteriophage fd (Otsuka and
                     Kunisawa, 1982)"
     rep_origin      1295..1750
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     rep_origin      complement(1909..2497)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     regulatory      complement(2595..2622)
                     /gene="trpA"
                     /regulatory_class="terminator"
     gene            complement(2595..2622)
                     /gene="trpA"
                     /label=trpA
     CDS             complement(2646..3437)
                     /label=NeoR/KanR
                     /note="aminoglycoside phosphotransferase"
     protein_bind    complement(3829..3850)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     CDS             complement(3866..4945)
                     /label=lacI
                     /note="lac repressor"
     promoter        complement(4946..5023)
                     /label=lacIq promoter
                     /note="In the lacIq allele, a single base change in the
                     promoter boosts expression of the lacI gene about 10-fold."
ORIGIN
        1 gatctcgatc ccgcgaaatt aatacgactc actatagggg aattgtgagc ggataacaat
       61 tcccctctag cgcgtattca ggctgaccct gcgcgctgcg cagggcttta ttgattccat
      121 ttttacactg atgaatgttc cgttgcgctg cccggattac aggggaatta attctcactc
      181 attaggcacc ccaggcttta cactttatgc ttccggctcg tataatgtgt ggaattgtga
      241 gcggataaca atttcacaca ggaaacagaa ttaattctta cacttagtta aattgctaac
      301 tttatagatt acaaaactta ggagggtttt taccatgggg gatcccaagc ttcttctaga
      361 ggtaccgcat gcgatatcga gctctcccgg gaattcactg gccgtcgttt tacaacgtcg
      421 tgactgggaa aaccctggcg ttacccaact taatcgcctt gcagcacatc cccctttcgc
      481 cagctggcgt aatagcgaag aggcccgcac cgatcgccct tcccaacagt tgcgcagcct
      541 gatccggctg ctaacaaagc ccgaaaggaa gctgagttgg ctgctgccac cgctgagcaa
      601 taactagcat aaccccttgg ggcctctaaa cgggtcttga ggggtttttt gctgaaagga
      661 ggaactatat ccggatatgc gagagtaggg aactgccagg catcaaataa aacgaaaggc
      721 tcagtcgaaa gactgggcct ttcgttttat ctgttgtttg tcggtgaacg ctctcctgag
      781 taggacaaat ccgccgggag cggatttgaa cgttgcgaag caacggcccg gagggtggcg
      841 ggcaggacgc ccgccataaa ctgccaggca tcaaattaag cagaaggcca tcctgacgga
      901 tggccttttt gcgtttctac aaactctttt gtttattttt ctaaatacat tcaaatatgt
      961 atccgctcat gctagaggtc gaaattcacc tcgaaagcaa gctgataaac cgatacaatt
     1021 aaaggctcct tttggagcct ttttttttgg agattttcaa cgtgaaaaaa ttattattcg
     1081 caattccaag ctaattcacc tcgaaagcaa gctgataaac cgatacaatt aaaggctcct
     1141 tttggagcct ttttttttgg agattttcaa cgtgaaaaaa ttattattcg caattccaag
     1201 ctctgcctcg cgcgtttcgg tgatgacggt gaaaacctct gacacatgca gctcccggag
     1261 acggtcacag cttgtctgta agcggatgca gatcacgcgc cctgtagcgg cgcattaagc
     1321 gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc cctagcgccc
     1381 gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccagctttcc ccgtcaagct
     1441 ctaaatcggg ggctcccttt agggttccga tttagtgctt tacggcacct cgaccccaaa
     1501 aaacttgatt agggtgatgg ttcacgtagt gggccatcgc cctgatagac ggtttttcgc
     1561 cctttgacgt tggagtccac gttctttaat agtggactct tgttccaaac tggaacaaca
     1621 ctcaacccta tctcggtcta ttcttttgat ttataaggga ttttgccgat ttcggcctat
     1681 tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attttaacaa aatattaacg
     1741 tttacaattt gatctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa
     1801 ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa
     1861 aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct
     1921 ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac
     1981 aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc
     2041 gaccctgccg cttaccggat acctctccgc ctttctccct tcgggaagcg tggcgctttc
     2101 tcaatgctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg
     2161 tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga
     2221 gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta acaggattag
     2281 cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta
     2341 cactagaagg acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag
     2401 agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg
     2461 caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac
     2521 ggggtctgac gctgactgga acgaaaactc acgttaaggg attttggtca tgtcgacctg
     2581 caggcatgca agctaaaaaa aagcccgctc attaggcggg ctagcgaacc ccagagtccc
     2641 gctcagaaga actcgtcaag aaggcgatag aaggcgatgc gctgcgaatc gggagcggcg
     2701 ataccgtaaa gcacgaggaa gcggtcagcc cattcgccgc caagctcttc agcaatatca
     2761 cgggtagcca acgctatgtc ctgatagcgg tccgccacac ccagccggcc acagtcgatg
     2821 aatccagaaa agcggccatt ttccaccatg atattcggca agcaggcatc gccatgggtc
     2881 acgacgagat cctcgccgtc gggcatgcgc gccttgagcc tggcgaacag ttcggctggc
     2941 gcgagcccct gatgctcttc gtccagatca tcctgatcga caagaccggc ttccatccga
     3001 gtacgtgctc gctcgatgcg atgtttcgct tggtggtcga atgggcaggt agccggatca
     3061 agcgtatgca gccgccgcat tgcatcagcc atgatggata ctttctcggc aggagcaagg
     3121 tgagatgaca ggagatcctg ccccggcact tcgcccaata gcagccagtc ccttcccgct
     3181 tcagtgacaa cgtcgagcac agctgcgcaa ggaacgcccg tcgtggccag ccacgatagc
     3241 cgcgctgcct cgtcctgcag ttcattcagg gcaccggaca ggtcggtctt gacaaaaaga
     3301 accgggcgcc cctgcgctga cagccggaac acggcggcat cagagcagcc gattgtctgt
     3361 tgtgcccagt catagccgaa tagcctctcc acccaagcgg ccggagaacc tgcgtgcaat
     3421 ccatcttgtt caatcatgcg aaacgatcct catcctgtct cttgatcaga tcttgatccc
     3481 ctgcgccatc agatccttgg cggcaagaaa gccatccagt ttactttgca gggcttccca
     3541 accttaccag agggcgcccc agctggcaat tccggttcgc ttgctgtcca taaaaccgcc
     3601 cagtctagct atcgccatgt aagcccactg caagctacct gctttctctt tgcgcttcgc
     3661 ttttcccttg tccagatagc ccagtagctg acattcatcc ggggtcagca ccgtttctgc
     3721 ggactggctt tctacgtgtt ccgcttcctt tagcagccct tgcgccctga gtgcttgcgg
     3781 cagcgtgaag ctggtcggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta
     3841 actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca
     3901 gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgccaggg
     3961 tggtttttct tttcaccagt gagacgggca acagctgatt gcccttcacc gcctggccct
     4021 gagagagttg cagcaagcgg tccacgctgg tttgccccag caggcgaaaa tcctgtttga
     4081 tggtggttga cggcgggata taacatgagc tgtcttcggt atcgtcgtat cccactaccg
     4141 agatatccgc accaacgcgc agcccggact cggtaatggc gcgcattgcg cccagcgcca
     4201 tctgatcgtt ggcaaccagc atcgcagtgg gaacgatgcc ctcattcagc atttgcatgg
     4261 tttgttgaaa accggacatg gcactccagt cgccttcccg ttccgctatc ggctgaattt
     4321 gattgcgagt gagatattta tgccagccag ccagacgcag acgcgccgag acagaactta
     4381 atgggcccgc taacagcgcg atttgctggt gacccaatgc gaccagatgc tccacgccca
     4441 gtcgcgtacc gtcttcatgg gagaaaataa tactgttgat gggtgtctgg tcagagacat
     4501 caagaaataa cgccggaaca ttagtgcagg cagcttccac agcaatggca tcctggtcat
     4561 ccagcggata gttaatgatc agcccactga cgcgttgcgc gagaagattg tgcaccgccg
     4621 ctttacaggc ttcgacgccg cttcgttcta ccatcgacac caccacgctg gcacccagtt
     4681 gatcggcgcg agatttaatc gccgcgacaa tttgcgacgg cgcgtgcagg gccagactgg
     4741 aggtggcaac gccaatcagc aacgactgtt tgcccgccag ttgttgtgcc acgcggttgg
     4801 gaatgtaatt cagctccgcc atcgccgctt ccactttttc ccgcgttttc gcagaaacgt
     4861 ggctggcctg gttcaccacg cgggaaacgg tctgataaga gacaccggca tactctgcga
     4921 catcgtataa cgttactggt ttcacattca ccaccctgaa ttgactctct tccgggcgct
     4981 atcatgccat accgcgaaag gttttgcacc attctatggt gtcgggtacc gagctcgaat
     5041 t
//
