LOCUS       40924_27019        3741 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pKSM711, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3741)
  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 3741)
  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 3741)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 3741)
  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, Cellular and Developmental 
            Biology, Harvard University, 16 Divinity Avenue, Cambridge, MA 
            02138, USA"
COMMENT     SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..3741
                     /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"
     CDS             complement(2595..3386)
                     /label=NeoR/KanR
                     /note="aminoglycoside phosphotransferase"
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 tgtcgaaccc
     2581 cagagtcccg ctcagaagaa ctcgtcaaga aggcgataga aggcgatgcg ctgcgaatcg
     2641 ggagcggcga taccgtaaag cacgaggaag cggtcagccc attcgccgcc aagctcttca
     2701 gcaatatcac gggtagccaa cgctatgtcc tgatagcggt ccgccacacc cagccggcca
     2761 cagtcgatga atccagaaaa gcggccattt tccaccatga tattcggcaa gcaggcatcg
     2821 ccatgggtca cgacgagatc ctcgccgtcg ggcatgcgcg ccttgagcct ggcgaacagt
     2881 tcggctggcg cgagcccctg atgctcttcg tccagatcat cctgatcgac aagaccggct
     2941 tccatccgag tacgtgctcg ctcgatgcga tgtttcgctt ggtggtcgaa tgggcaggta
     3001 gccggatcaa gcgtatgcag ccgccgcatt gcatcagcca tgatggatac tttctcggca
     3061 ggagcaaggt gagatgacag gagatcctgc cccggcactt cgcccaatag cagccagtcc
     3121 cttcccgctt cagtgacaac gtcgagcaca gctgcgcaag gaacgcccgt cgtggccagc
     3181 cacgatagcc gcgctgcctc gtcctgcagt tcattcaggg caccggacag gtcggtcttg
     3241 acaaaaagaa ccgggcgccc ctgcgctgac agccggaaca cggcggcatc agagcagccg
     3301 attgtctgtt gtgcccagtc atagccgaat agcctctcca cccaagcggc cggagaacct
     3361 gcgtgcaatc catcttgttc aatcatgcga aacgatcctc atcctgtctc ttgatcagat
     3421 cttgatcccc tgcgccatca gatccttggc ggcaagaaag ccatccagtt tactttgcag
     3481 ggcttcccaa ccttaccaga gggcgcccca gctggcaatt ccggttcgct tgctgtccat
     3541 aaaaccgccc agtctagcta tcgccatgta agcccactgc aagctacctg ctttctcttt
     3601 gcgcttcgct tttcccttgt ccagatagcc cagtagctga cattcatccg gggtcagcac
     3661 cgtttctgcg gactggcttt ctacgtgttc cgcttccttt agcagccctt gcgccctgag
     3721 tgcttgcggc agcgtgaagc t
//
