LOCUS LC459971 2556 bp DNA circular SYN 25-APR-2019 DEFINITION Cloning vector pIIS18-SapI DNA, complete sequence. ACCESSION LC459971 VERSION LC459971.1 KEYWORDS . SOURCE Cloning vector pIIS18-SapI ORGANISM Cloning vector pIIS18-SapI other sequences; artificial sequences; vectors. REFERENCE 1 (bases 1 to 2556) 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TITLE A new E. coli entry vector series named pIIS18 for seamless cloning using Type IIS restriction enzymes JOURNAL Unpublished (2018) COMMENT FEATURES Location/Qualifiers source 1..2556 /db_xref="taxon:2510005" /mol_type="other DNA" /organism="Cloning vector pIIS18-SapI" CDS complement(61..390) /codon_start=1 /gene="lacZ'" /product="lacZ alpha fragment of beta-galactosidase" /protein_id="BBI37008.1" /transl_table=11 /translation="MTMITNSSSVPGDPRSSRRACRHASLALAVVLQRRDWENPGVTQ LNRLAAHPPFASWRNSEEARTDRPSQQLRSLNGEWRLMRYFLLTHLCGISHRIWCTLS TICSDAA" misc_feature 314..376 /note="modified multiple cloning site to include two facing TypeIIS cleavage sites overlapped by one blunt ended cleavage site" misc_feature 332..338 /note="SapI cleavage site (TypeIIS cleavage site)" misc_feature 337..342 /note="Bsp68I cleavage site (Blunt end enzyme cleavage site)" misc_feature complement(341..347) /note="SapI cleavage site(TypeIIS cleavage site)" protein_bind complement(410..426) /bound_moiety="lac repressor encoded by lacI" /gene="lacO" protein_bind complement(479..500) /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." rep_origin complement(788..1376) /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" CDS complement(1547..2407) /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="BBI37009.1" /transl_table=11 /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGY IELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVE YSPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRL DRWEPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPL LRSALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIA EIGASLIKHW" BASE COUNT 636 a 642 c 652 g 626 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 ggctcttcgc gaagagcgag gatccccggg 361 taccgagctc gaattcgtaa tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc 421 tcacaattcc acacaacata cgagccggaa gcataaagtg taaagcctgg ggtgcctaat 481 gagtgagcta actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc 541 tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg 601 ggcgctattc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag 661 cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag 721 gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc 781 tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc 841 agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc 901 tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt 961 cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg 1021 ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat 1081 ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag 1141 ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt 1201 ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc 1261 cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta 1321 gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag 1381 atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga 1441 ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa 1501 gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa 1561 tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc 1621 ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga 1681 taccgcgaga gccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa 1741 gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt 1801 gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg 1861 ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc 1921 aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg 1981 gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag 2041 cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt 2101 actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt 2161 caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac 2221 gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac 2281 ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag 2341 caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa 2401 tactcatact cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga 2461 gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc 2521 cccgaaaagt gccacctgac gtctaagaaa ccatta //