LOCUS LC459975 2562 bp DNA circular SYN 25-APR-2019 DEFINITION Cloning vector pIIS18-BfuAI-2 DNA, complete sequence. ACCESSION LC459975 VERSION LC459975.1 KEYWORDS . SOURCE Cloning vector pIIS18-BfuAI-2 ORGANISM Cloning vector pIIS18-BfuAI-2 other sequences; artificial sequences; vectors. REFERENCE 1 (bases 1 to 2562) AUTHORS Altaib,H.E. and Suzuki,T. TITLE Direct Submission JOURNAL Submitted (24-JAN-2019) to the DDBJ/EMBL/GenBank databases. Contact:Hend Essam Altaib Gifu University, Faculty of Applied Biological Sciences; 1-1 yanagido, Gifu, Gifu prefecture 501-1193, Japan REFERENCE 2 AUTHORS Altaib,H., Takiguchi,Y., Kozakai,T., Badr,Y., Nomura,I. and Suzuki,T. TITLE A new E. coli entry vector series named pIIS18 for seamless cloning using Type IIS restriction enzymes JOURNAL Unpublished (2018) COMMENT FEATURES Location/Qualifiers source 1..2562 /db_xref="taxon:2510009" /mol_type="other DNA" /organism="Cloning vector pIIS18-BfuAI-2" CDS complement(61..396) /codon_start=1 /gene="LacZ'" /product="lacZ alpha fragment of beta glactosidase" /protein_id="BBI37016.1" /transl_table=11 /translation="MTMITNSSSVPGDPHLHLNAGGCRHASLALAVVLQRRDWENPGV TQLNRLAAHPPFASWRNSEEARTDRPSQQLRSLNGEWRLMRYFLLTHLCGISHRIWCT LSTICSDAA" 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 /note="BfuAI cleavage site(TypeIIS cleavage site)" misc_feature 340..347 /note="SwaI cleavage site(Blunt end enzyme cleavage site)" misc_feature complement(348..353) /note="BfuAI cleavage site(TypeIIS cleavage site)" protein_bind complement(416..432) /bound_moiety="lac repressor encoded by lacI" /gene="lacO" protein_bind complement(485..506) /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." rep_origin complement(794..1382) /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" CDS complement(1553..2413) /codon_start=1 /gene="bla(AmpR)" /note="confers resistance to ampicillin, carbenicillin, and related antibiotics.It has single nucleotide change(with no effect on function)to remove unnecessary TypeIIS BsaI cleavage site." /product="beta-lactamase" /protein_id="BBI37017.1" /transl_table=11 /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGY IELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVE YSPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRL DRWEPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPL LRSALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIA EIGASLIKHW" BASE COUNT 638 a 644 c 651 g 629 t 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 //