LOCUS       62056_12840        2559 bp DNA     circular SYN 25-APR-2019
DEFINITION  Cloning vector pIIS18-BfuAI-1 DNA, complete sequence.
ACCESSION   LC459974
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2559)
  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  JOURNAL   Unpublished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 2559)
  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 2559)
  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            "Unpublished"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..2559
                     /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..379
                     /note="modified multiple cloning site to include two facing
                     TypeIIS cleavage sites overlapped by one blunt ended 
                     cleavage site."
     misc_feature    333..338
                     /label=BfuAI cleavage site(TypeIIS cleavage site)
                     /note="BfuAI cleavage site(TypeIIS cleavage site)"
     misc_feature    340..345
                     /note="EcoRV cleavage site(Blunt end enzyme cleavage site)"
     misc_feature    complement(347..352)
                     /gene="lacZ'"
                     /label=BfuAI cleavage site(TypeIIS cleavage site)
                     /note="BfuAI cleavage site(TypeIIS cleavage site)"
     primer_bind     complement(389..405)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(413..429)
                     /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(437..467)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(482..503)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(791..1379)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(1553..2410)
                     /label=AmpR
                     /note="beta-lactamase"
     promoter        complement(2411..2515)
                     /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 gcacctgcag atatctgcag gtggatcccc
      361 gggtaccgag ctcgaattcg taatcatggt catagctgtt tcctgtgtga aattgttatc
      421 cgctcacaat tccacacaac atacgagccg gaagcataaa gtgtaaagcc tggggtgcct
      481 aatgagtgag ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa
      541 acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta
      601 ttgggcgcta ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc
      661 gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg
      721 caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt
      781 tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa
      841 gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct
      901 ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc
      961 cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg
     1021 tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct
     1081 tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag
     1141 cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga
     1201 agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga
     1261 agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg
     1321 gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag
     1381 aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag
     1441 ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat
     1501 gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct
     1561 taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac
     1621 tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa
     1681 tgataccgcg agagccacgc tcaccggctc cagatttatc agcaataaac cagccagccg
     1741 gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt
     1801 gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca
     1861 ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt
     1921 cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct
     1981 tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg
     2041 cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg
     2101 agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg
     2161 cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa
     2221 aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt
     2281 aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt
     2341 gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt
     2401 gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca
     2461 tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat
     2521 ttccccgaaa agtgccacct gacgtctaag aaaccatta
//
