LOCUS Exported 3226 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION C-terminal Pk6-tag. ACCESSION . VERSION . KEYWORDS pJet1.2 Pk6-tag SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 3226) 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 3226) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..3226 /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(1058..1088) /label=lac UV5 promoter /note="E. coli lac promoter with an ""up"" mutation" protein_bind 1103..1124 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(1244..1261) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(1415..2003) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(1495..1514) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(2174..3034) /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 2797..2816 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(3035..3139) /gene="bla" /label=AmpR promoter primer_bind 3207..3225 /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 ctcaagcggt tctgctgcta gaggtattcc taacccttta 421 ttgggattag atggaatccc aaatcctttg ctaggtcttg acggcatacc caatccatta 481 ttaggcttgg atggatcctc taggggtatt cctaaccctt tattgggatt agatggaatc 541 ccaaatcctt tgctaggtct tgacggcata cccaatccat tattaggctt ggatggatcc 601 tctagctaag tgagacccaa tgaatctttc tagaagatct cctacaatat tctcagctgc 661 catggaaaat cgatgttctt cttttattct ctcaagattt tcaggctgta tattaaaact 721 tatattaaga actatgctaa ccacctcatc aggaaccgtt gtaggtggcg tgggttttct 781 tggcaatcga ctctcatgaa aactacgagc taaatattca atatgttcct cttgaccaac 841 tttattctgc attttttttg aacgaggttt agagcaagct tcaggaaact gagacaggaa 901 ttttattaaa aatttaaatt ttgaagaaag ttcagggtta atagcatcca ttttttgctt 961 tgcaagttcc tcagcattct taacaaaaga cgtctctttt gacatgttta aagtttaaac 1021 ctcctgtgtg aaattattat ccgctcataa ttccacacat tatacgagcc ggaagcataa 1081 agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac 1141 tgccaattgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac 1201 gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc 1261 tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt 1321 tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg 1381 ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg 1441 agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat 1501 accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta 1561 ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct 1621 gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc 1681 ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa 1741 gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg 1801 taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag 1861 tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt 1921 gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta 1981 cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc 2041 agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca 2101 cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa 2161 cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat 2221 ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct 2281 taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt 2341 tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat 2401 ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta 2461 atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg 2521 gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt 2581 tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg 2641 cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg 2701 taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc 2761 ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa 2821 ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac 2881 cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt 2941 ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg 3001 gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa 3061 gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata 3121 aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc taagaaacca 3181 ttattatcat gacattaacc tataaaaata ggcgtatcac gaggcc //