LOCUS Exported 4009 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Spectinomycin resistance cassette. ACCESSION . VERSION . KEYWORDS pJet1.2 specR cassette SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 4009) AUTHORS Diebold-Durand ML, Burmann F, Gruber S TITLE High-Throughput Allelic Replacement Screening in Bacillus subtilis. JOURNAL Methods Mol Biol. 2019;2004:49-61. doi: 10.1007/978-1-4939-9520-2_5. PUBMED 31147909 REFERENCE 2 (bases 1 to 4009) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..4009 /organism="synthetic DNA construct" /mol_type="other DNA" primer_bind 305..324 /label=T7 /note="T7 promoter, forward primer" promoter 305..323 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" promoter complement(1841..1871) /label=lac UV5 promoter /note="E. coli lac promoter with an ""up"" mutation" protein_bind 1886..1907 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(2027..2044) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(2198..2786) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(2278..2297) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(2957..3817) /codon_start=1 /gene="bla" /product="beta-lactamase" /label=AmpR /note="confers resistance to ampicillin, carbenicillin, and related antibiotics" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS LIKHW" primer_bind 3580..3599 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(3818..3922) /gene="bla" /label=AmpR promoter primer_bind 3990..4008 /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" ORIGIN 1 gcccctgcag ccgaattata ttatttttgc caaataattt ttaacaaaag ctctgaagtc 61 ttcttcattt aaattcttag atgatacttc atctggaaaa ttgtcccaat tagtagcatc 121 acgctgtgag taagttctaa accatttttt tattgttgta ttatctctaa tcttactact 181 cgatgagttt tcggtattat ctctattttt aacttggagc aggttccatt cattgttttt 241 ttcatcatag tgaataaaat caactgcttt aacacttgtg cctgaacacc atatccatcc 301 ggcgtaatac gactcactat agggagagcg gccgccagat cttccggatg gctcgagttt 361 ttcagcaaga tgttacaggt ctcgaatatc ttcttacttc tgtataataa agaataatta 421 ttaatctgta gacaaattgt gaaaggatgt acttaaacgc taacggtcag ctttattgaa 481 cagtaattta agtatatgtc caatctaggg taagtaaatt gagtatcaat ataaacttta 541 tatgaacata atcaacgagg tgaaatcatg agcaatttga ttaacggaaa aataccaaat 601 caagcgattc aaacattaaa aatcgtaaaa gatttatttg gaagttcaat agttggagta 661 tatctatttg gttcagcagt aaatggtggt ttacgcatta acagcgatgt agatgttcta 721 gtcgtcgtga atcatagttt acctcaatta actcgaaaaa aactaacaga aagactaatg 781 actatatcag gaaagattgg aaatacggat tctgttagac cacttgaagt tacggttata 841 aataggagtg aagttgtccc ttggcaatat cctccaaaaa gagaatttat atacggtgag 901 tggctcaggg gtgaatttga gaatggacaa attcaggaac caagctatga tcctgatttg 961 gctattgttt tagcacaagc aagaaagaat agtatttctc tatttggtcc tgattcttca 1021 agtatacttg tctccgtacc tttgacagat attcgaagag caattaagga ttctttgcca 1081 gaactaattg aggggataaa aggtgatgag cgtaatgtaa ttttaaccct agctcgaatg 1141 tggcaaacag tgactactgg tgaaattacc tcgaaagatg tcgctgcaga atgggctata 1201 cctcttttac ctaaagagca tgtaacttta ctggatatag ctagaaaagg ctatcgggga 1261 gagtgtgatg ataagtggga aggactatat tcaaaggtga aagcactcgt taagtatatg 1321 aaaaattcta tagaaacttc tctcaattaa gaaaaaggcc ccaacatctt ggggcctttt 1381 tcttttttat ctacgagacc caatgaatct ttctagaaga tctcctacaa tattctcagc 1441 tgccatggaa aatcgatgtt cttcttttat tctctcaaga ttttcaggct gtatattaaa 1501 acttatatta agaactatgc taaccacctc atcaggaacc gttgtaggtg gcgtgggttt 1561 tcttggcaat cgactctcat gaaaactacg agctaaatat tcaatatgtt cctcttgacc 1621 aactttattc tgcatttttt ttgaacgagg tttagagcaa gcttcaggaa actgagacag 1681 gaattttatt aaaaatttaa attttgaaga aagttcaggg ttaatagcat ccattttttg 1741 ctttgcaagt tcctcagcat tcttaacaaa agacgtctct tttgacatgt ttaaagttta 1801 aacctcctgt gtgaaattat tatccgctca taattccaca cattatacga gccggaagca 1861 taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct 1921 cactgccaat tgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc 1981 aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact 2041 cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac 2101 ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa 2161 aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg 2221 acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa 2281 gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc 2341 ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac 2401 gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac 2461 cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg 2521 taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt 2581 atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagga 2641 cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct 2701 cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga 2761 ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg 2821 ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct 2881 tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt 2941 aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc 3001 tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg 3061 gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag 3121 atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt 3181 tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag 3241 ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt 3301 ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca 3361 tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg 3421 ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat 3481 ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta 3541 tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca 3601 gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct 3661 taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat 3721 cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa 3781 agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt 3841 gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa 3901 ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac gtctaagaaa 3961 ccattattat catgacatta acctataaaa ataggcgtat cacgaggcc //