LOCUS       62056_12845        2562 bp DNA     circular SYN 25-APR-2019
DEFINITION  Cloning vector pIIS18-BfuAI-2 DNA, complete sequence.
ACCESSION   LC459975
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2562)
  AUTHORS   Altaib H, Takiguchi Y, Kozakai T, Badr Y, Nomura I, Suzuki T.
  TITLE     A new E. coli entry vector series named pIIS18 for seamless cloning 
            using Type IIS restriction enzymes
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 2562)
  AUTHORS   Altaib HE, Suzuki T.
  TITLE     Direct Submission
  JOURNAL   Submitted (24-JAN-2019) Contact:Hend Essam Altaib Gifu University, 
            Faculty of Applied Biological Sciences; 1-1 yanagido, Gifu, Gifu 
            prefecture 501-1193, Japan
REFERENCE   3  (bases 1 to 2562)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (24-JAN-2019) Contact:Hend Essam Altaib Gifu University, Faculty of 
            Applied Biological Sciences; 1-1 yanagido, Gifu, Gifu prefecture 
            501-1193, Japan"
FEATURES             Location/Qualifiers
     source          1..2562
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     294..310
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    314..382
                     /note="modified multiple cloning site to include two facing
                     TypeIIS cleavage sites overlapped by one blunt ended 
                     cleavage site."
     misc_feature    334..339
                     /label=BfuAI cleavage site(TypeIIS cleavage site)
                     /note="BfuAI cleavage site(TypeIIS cleavage site)"
     misc_feature    340..347
                     /label=SwaI cleavage site(Blunt end enzyme cleavage site)
                     /note="SwaI cleavage site(Blunt end enzyme cleavage site)"
     misc_feature    complement(348..353)
                     /gene="LacZ'"
                     /label=BfuAI cleavage site(TypeIIS cleavage site)
                     /note="BfuAI cleavage site(TypeIIS cleavage site)"
     primer_bind     complement(392..408)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(416..432)
                     /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        complement(440..470)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(485..506)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(794..1382)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(1556..2413)
                     /label=AmpR
                     /note="beta-lactamase"
     promoter        complement(2414..2518)
                     /label=AmpR promoter
ORIGIN
        1 gagcagacaa gcccgtcagg gcgcgtcagc gggtgttggc gggtgtcggg gctggcttaa
       61 ctatgcggca tcagagcaga ttgtactgag agtgcaccat atgcggtgtg aaataccgca
      121 cagatgcgta aggagaaaat accgcatcag gcgccattcg ccattcaggc tgcgcaactg
      181 ttgggaaggg cgatcggtgc gggcctcttc gctattacgc cagctggcga aagggggatg
      241 tgctgcaagg cgattaagtt gggtaacgcc agggttttcc cagtcacgac gttgtaaaac
      301 gacggccagt gccaagcttg catgcctgca gccacctgca tttaaatgca ggtgcggatc
      361 cccgggtacc gagctcgaat tcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt
      421 atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg
      481 cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg
      541 gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc
      601 gtattgggcg ctattccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc
      661 ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata
      721 acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg
      781 cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct
      841 caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa
      901 gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc
      961 tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt
     1021 aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg
     1081 ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg
     1141 cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct
     1201 tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc
     1261 tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg
     1321 ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc
     1381 aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt
     1441 aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa
     1501 aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat
     1561 gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct
     1621 gactccccgt cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg
     1681 caatgatacc gcgagagcca cgctcaccgg ctccagattt atcagcaata aaccagccag
     1741 ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta
     1801 attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg
     1861 ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg
     1921 gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct
     1981 ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta
     2041 tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg
     2101 gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc
     2161 cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg
     2221 gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga
     2281 tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg
     2341 ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat
     2401 gttgaatact catactcttc ctttttcaat attattgaag catttatcag ggttattgtc
     2461 tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca
     2521 catttccccg aaaagtgcca cctgacgtct aagaaaccat ta
//
