LOCUS Exported 3922 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION C-terminal Tev Halo-Tag. ACCESSION . VERSION . KEYWORDS pJet1.2 Tev-Halo tag SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 3922) 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 3922) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..3922 /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" CDS complement(383..1267) /codon_start=1 /product="modified bacterial dehalogenase that forms covalent bonds with chloroalkane derivatives" /label=HaloTag(R) /translation="EIGTGFPFDPHYVEVLGERMHYVDVGPRDGTPVLFLHGNPTSSYV WRNIIPHVAPTHRCIAPDLIGMGKSDKPDLGYFFDDHVRFMDAFIEALGLEEVVLVIHD WGSALGFHWAKRNPERVKGIAFMEFIRPIPTWDEWPEFARETFQAFRTTDVGRKLIIDQ NVFIEGTLPMGVVRPLTEVEMDHYREPFLNPVDREPLWRFPNELPIAGEPANIVALVEE YMDWLHQSPVPKLLFWGTPGVLIPPAEAARLAKSLPNCKAVDIGPGLNLLQEDNPDLIG SEIARWLSTLEISG" CDS complement(1280..1300) /codon_start=1 /product="tobacco etch virus (TEV) protease recognition and cleavage site" /label=TEV site /translation="ENLYFQG" promoter complement(1754..1784) /label=lac UV5 promoter /note="E. coli lac promoter with an ""up"" mutation" protein_bind 1799..1820 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(1940..1957) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(2111..2699) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(2191..2210) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(2870..3730) /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 3493..3512 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(3731..3835) /gene="bla" /label=AmpR promoter primer_bind 3903..3921 /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 tggtctcgct taaccggaaa tctccagagt agacagccag cgcgcgatct 421 cgctgccgat caggtccggg ttgtcttctt gcagcagatt cagacccggg ccgatgtcca 481 cagccttgca gttaggcagg cttttggcca ggcgagcggc ttcggccggt gggatcagaa 541 cgcctggggt gccccagaac agcagcttcg ggacagggga ctggtgcagc cagtccatgt 601 attcttcgac cagcgcgacg atgttcgctg gctcaccggc gattggcagc tcgtttggga 661 agcgccacag tggctcgcgg tcaacaggat tcaggaacgg ctcgcggtaa tggtccatct 721 cgacttcagt cagcgggcgg acgacaccca tcggcagcgt accctcgata aaaacgttct 781 gatcgatgat cagcttgcgg ccgacgtcgg tggtgcggaa ggcctggaag gtttcgcggg 841 caaattctgg ccattcgtcc caggtcggga tagggcggat gaactccata aatgcaatac 901 ctttgacgcg ctctggattg cgcttggccc agtggaaacc cagagcggag ccccagtcgt 961 gaatgaccag gacgacctct tccagaccca gggcttcgat gaaggcatcc atgaagcgga 1021 cgtggtcgtc gaagaaataa cccaggtctg gtttgtcgga tttgcccata ccgatcaggt 1081 ctggagcaat gcagcgatgg gtcggtgcaa catgcgggat gatgttgcgc cacacgtagg 1141 aggaggtcgg gttaccgtgc aggaacagca caggggtgcc atcgcgcgga ccaacatcga 1201 cgtagtgcat gcgctcgccc aggacttcca cataatgggg gtcgaatgga aagccagtac 1261 cgatttcgct tccgctagca ccctgaaaat acaaattctc tccgcttccg ctcgagacca 1321 tctttctaga agatctccta caatattctc agctgccatg gaaaatcgat gttcttcttt 1381 tattctctca agattttcag gctgtatatt aaaacttata ttaagaacta tgctaaccac 1441 ctcatcagga accgttgtag gtggcgtggg ttttcttggc aatcgactct catgaaaact 1501 acgagctaaa tattcaatat gttcctcttg accaacttta ttctgcattt tttttgaacg 1561 aggtttagag caagcttcag gaaactgaga caggaatttt attaaaaatt taaattttga 1621 agaaagttca gggttaatag catccatttt ttgctttgca agttcctcag cattcttaac 1681 aaaagacgtc tcttttgaca tgtttaaagt ttaaacctcc tgtgtgaaat tattatccgc 1741 tcataattcc acacattata cgagccggaa gcataaagtg taaagcctgg ggtgcctaat 1801 gagtgagcta actcacatta attgcgttgc gctcactgcc aattgctttc cagtcgggaa 1861 acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta 1921 ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc 1981 gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg 2041 caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt 2101 tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa 2161 gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct 2221 ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc 2281 cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg 2341 tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct 2401 tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag 2461 cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga 2521 agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga 2581 agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg 2641 gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag 2701 aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag 2761 ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat 2821 gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct 2881 taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac 2941 tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa 3001 tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg 3061 gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt 3121 gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca 3181 ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt 3241 cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct 3301 tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg 3361 cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg 3421 agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg 3481 cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa 3541 aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt 3601 aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt 3661 gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt 3721 gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca 3781 tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat 3841 ttccccgaaa agtgccacct gacgtctaag aaaccattat tatcatgaca ttaacctata 3901 aaaataggcg tatcacgagg cc //