LOCUS       40924_27014        5071 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pKSM710, complete sequence.
ACCESSION   .
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)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 5071)
  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..5071
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     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
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     rep_origin      complement(927..1515)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(1689..2546)
                     /label=AmpR
                     /note="beta-lactamase"
     regulatory      complement(2720..2774)
                     /regulatory_class="terminator"
     CDS             complement(2864..3520)
                     /label=CmR
                     /note="chloramphenicol acetyltransferase"
     promoter        complement(3521..3623)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding
                     chloramphenicol acetyltransferase"
     regulatory      3881..3910
                     /label=lambda Pl promoter
                     /note="lambda Pl promoter"
                     /regulatory_class="promoter"
     promoter        4109..4127
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    4128..4152
                     /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        4286..4316
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an 'up' mutation"
     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"
     terminator      4793..4879
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     terminator      4971..4998
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB
                     gene"
ORIGIN
        1 aattcacctc gaaagcaagc tgataaaccg atacaattaa aggctccttt tggagccttt
       61 ttttttggag attttcaacg tgaaaaaatt attattcgca attccaagct aattcacctc
      121 gaaagcaagc tgataaaccg atacaattaa aggctccttt tggagccttt ttttttggag
      181 attttcaacg tgaaaaaatt attattcgca attccaagct ctgcctcgcg cgtttcggtg
      241 atgacggtga aaacctctga cacatgcagc tcccggagac ggtcacagct tgtctgtaag
      301 cggatgcaga tcacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc
      361 gcagcgtgac cgctacactt gccagcgccc tagcgcccgc tcctttcgct ttcttccctt
      421 cctttctcgc cacgttcgcc agctttcccc gtcaagctct aaatcggggg ctccctttag
      481 ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgattag ggtgatggtt
      541 cacgtagtgg gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt
      601 tctttaatag tggactcttg ttccaaactg gaacaacact caaccctatc tcggtctatt
      661 cttttgattt ataagggatt ttgccgattt cggcctattg gttaaaaaat gagctgattt
      721 aacaaaaatt taacgcgaat tttaacaaaa tattaacgtt tacaatttga tctgcgctcg
      781 gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca
      841 gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac
      901 cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac
      961 aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg
     1021 tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac
     1081 ctctccgcct ttctcccttc gggaagcgtg gcgctttctc aatgctcacg ctgtaggtat
     1141 ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag
     1201 cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac
     1261 ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt
     1321 gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaaggac agtatttggt
     1381 atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc
     1441 aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga
     1501 aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tgactggaac
     1561 gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc
     1621 cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct
     1681 cacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca
     1741 tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct
     1801 ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca
     1861 ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc
     1921 atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg
     1981 cgcaacgttg ttgccattgc tgcaggcatc gtggtgtcac gctcgtcgtt tggtatggct
     2041 tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa
     2101 aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta
     2161 tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc
     2221 ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg
     2281 agttgctctt gcccggcgtc aacacgggat aataccgcgc cacatagcag aactttaaaa
     2341 gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg
     2401 agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc
     2461 accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg
     2521 gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg aagcagacag
     2581 ttttattgtt catgatgata tatttttatc ttgtgcaatg taacatcaga gattttgaga
     2641 aacaacgtgg ctttgttgaa taaatcgaac ttttgctgag ttgactcccc gcgcgcgatg
     2701 ggtcgaattt gctttcgaaa aaaaagcccg ctcattaggc gggctaaaaa aaagcccgct
     2761 cattaggcgg gctcgaattt ctgccattca tccgcttatt atcacttatt caggcgtagc
     2821 aaccaggcgt ttaagggcac caataagtgc cttaaaaaaa ttacgccccg ccctgccact
     2881 catcgcagta ctgttgtaat tcattaagca ttctgccgac atggaagcca tcacagacgg
     2941 catgatgaac ctgaatcgcc agcggcatca gcaccttgtc gccttgcgta taatatttgc
     3001 ccatagtgaa aacgggggcg aagaagttgt ccatattcgc cacgtttaaa tcaaaactgg
     3061 tgaaactcac ccagggattg gctgagacga aaaacatatt ctcaataaac cctttaggga
     3121 aataggccag gttttcaccg taacacgcca catcttgcga atatatgtgt agaaactgcc
     3181 ggaaatcgtc gtggtattca ctccagagcg atgaaaacgt ttcagtttgc tcatggaaaa
     3241 cggtgtaaca agggtgaaca ctatcccata tcaccagctc accgtctttc attgccatac
     3301 gaaattccgg atgagcattc atcaggcggg caagaatgtg aataaaggcc ggataaaact
     3361 tgtgcttatt tttctttacg gtctttaaaa aggccgtaat atccagctga acggtctggt
     3421 tataggtaca ttgagcaact gactgaaatg cctcaaaatg ttctttacga tgccattggg
     3481 atatatcaac ggtggtatat ccagtgattt ttttctccat tttagcttcc ttagctcctg
     3541 aaaatctcga taactcaaaa aatacgcccg gtagtgatct tatttcatta tggtgaaagt
     3601 tggaacctct tacgtgccga tcaacgtctc attttcgcca aaagttggcc cagggcttcc
     3661 cggtatcaac agggacacca ggatttattt attctgcgaa gtgatcttcc gtcacaggta
     3721 tttattcgaa gacgaaaggg catcgcgcgc ggggaattgg ccacgatgcg tccggcgtag
     3781 aggatctctc acctaccaaa caatgccccc ctgcaaaaaa taaattcata taaaaaacat
     3841 atagataacc atctgcggtg ataaattatc tctggcggtg ttgacataaa taccactggc
     3901 ggtgatactg agcacatcag caggacgcac tgaccaccat gaaggtgacg ctcttaaaat
     3961 taagccctga agaagggcag cattcaaagc agaaggcttt ggggtgtgtg atacgaaacg
     4021 aagcattgga attctacaac ttgcttggat tcctacaaag aagcagcaat tttcagtgtc
     4081 agaagtcgag atctcgatcc cgcgaaatta atacgactca ctatagggga attgtgagcg
     4141 gataacaatt cccctctagc gcgtattcag gctgaccctg cgcgctgcgc agggctttat
     4201 tgattccatt tttacactga tgaatgttcc gttgcgctgc ccggattaca ggggaattaa
     4261 ttctcactca ttaggcaccc caggctttac actttatgct tccggctcgt ataatgtgtg
     4321 gaattgtgag cggataacaa tttcacacag gaaacagaat taattcttac acttagttaa
     4381 attgctaact ttatagatta caaaacttag gagggttttt accatggggg atcccaagct
     4441 tcttctagag gtaccgcatg cgatatcgag ctctcccggg aattcactgg ccgtcgtttt
     4501 acaacgtcgt gactgggaaa accctggcgt tacccaactt aatcgccttg cagcacatcc
     4561 ccctttcgcc agctggcgta atagcgaaga ggcccgcacc gatcgccctt cccaacagtt
     4621 gcgcagcctg atccggctgc taacaaagcc cgaaaggaag ctgagttggc tgctgccacc
     4681 gctgagcaat aactagcata accccttggg gcctctaaac gggtcttgag gggttttttg
     4741 ctgaaaggag gaactatatc cggatatgcg agagtaggga actgccaggc atcaaataaa
     4801 acgaaaggct cagtcgaaag actgggcctt tcgttttatc tgttgtttgt cggtgaacgc
     4861 tctcctgagt aggacaaatc cgccgggagc ggatttgaac gttgcgaagc aacggcccgg
     4921 agggtggcgg gcaggacgcc cgccataaac tgccaggcat caaattaagc agaaggccat
     4981 cctgacggat ggcctttttg cgtttctaca aactcttttg tttatttttc taaatacatt
     5041 caaatatgta tccgctcatg ctagaggtcg a
//
