LOCUS Exported 4347 bp ds-DNA circular SYN 07-OCT-2021 DEFINITION Production of HaloTag fusion protein. ACCESSION . VERSION . KEYWORDS pH6HTC_YBX3-Halo SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 4347) TITLE Kutter Lab HaloTag plasmids 2021 REFERENCE 2 (bases 1 to 4347) AUTHORS . TITLE Direct Submission JOURNAL Exported Oct 7, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..4347 /organism="synthetic DNA construct" /mol_type="other DNA" terminator 55..141 /gene="Escherichia coli rrnB" /label=rrnB T1 terminator /note="transcription terminator T1 from the E. coli rrnB gene" terminator 233..260 /label=rrnB T2 terminator /note="transcription terminator T2 from the E. coli rrnB gene" primer_bind 342..361 /label=T7 /note="T7 promoter, forward primer" promoter 342..360 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" CDS 433..453 /codon_start=1 /product="6xHis affinity tag" /label=7xHis /translation="HHHHHHH" CDS 1334..1354 /codon_start=1 /product="tobacco etch virus (TEV) protease recognition and cleavage site" /label=TEV site /translation="EDLYFQS" CDS 1370..2254 /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 2255..2272 /codon_start=1 /product="6xHis affinity tag" /label=6xHis /translation="HHHHHH" primer_bind complement(2283..2301) /label=T7 Term /note="T7 terminator, reverse primer" terminator 2297..2344 /label=T7 terminator /note="transcription terminator for bacteriophage T7 RNA polymerase" promoter 2573..2677 /gene="bla" /label=AmpR promoter CDS 2678..3538 /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 complement(2896..2915) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin complement(3696..4284) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(3776..3795) /label=pBR322ori-F /note="pBR322 origin, forward primer" ORIGIN 1 catatgatgg tagtgtgggg tctccccatg cgagagtagg gaactgccag gcatcaaata 61 aaacgaaagg ctcagtcgaa agactgggcc tttcgtttta tctgttgttt gtcggtgaac 121 gctctcctga gtaggacaaa tccgccggga gcggatttga acgttgcgaa gcaacggccc 181 ggagggtggc gggcaggacg cccgccataa actgccaggc atcaaattaa gcagaaggcc 241 atcctgacgg atggcctttt tgcgtttcta caaactcttt tgtttatttt tctaaataca 301 ttcaaatatg tatccgctca tggatctcga tcccgcgaaa ttaatacgac tcactatagg 361 gtattttaac tttactaagg agcgatcgct tccgaattcc taccgcggat atatgagtga 421 ggcgggcgag gccaccacca ccaccaccac caccctcccg caggctccga cggaggcggc 481 cgccgcggct ccccaggacc ccgcgcccaa gagcccggtg ggcagcggtg cgccccaggc 541 cgcggccccg gcgcccgccg cccacgtcgc aggaaacccc ggtggggacg cggcccccgc 601 agccacgggc accgcggccg ccgcctcttt agccaccgcc gccggcagcg aagacgcgga 661 gaaaaaagtt ctcgccacca aagtccttgg cactgtcaaa tggttcaacg tcagaaatgg 721 atatggattt ataaatcgaa atgacaccaa agaagatgta tttgtacatc agactgccat 781 caagaagaat aacccacgga aatatctgcg cagtgtagga gatggagaaa ctgtagagtt 841 tgatgtggtt gaaggagaga agggtgcaga agctgccaat gtgactggcc cggatggagt 901 tcctgtggaa gggagtcgtt acgctgcaga tcggcgccgt tacagacgtg gctactatgg 961 aaggcgccgt ggccctcccc ggaatgctgg tgagattgga gagatgaagg atggagtccc 1021 agagggagca caacttcagg gaccggttca tcgaaatcca acttaccgcc caaggtaccg 1081 tagcagggga cctcctcgcc cacgacctgc cccagcagtt ggagaggctg aagataaaga 1141 aaatcagcaa gccaccagtg gtccaaacca gccgtctgtt cgccgtggat accggcgtcc 1201 ctacaattac cggcgtcgcc cgcgtcctcc taacgctcct tcacaagatg gcaaagaggc 1261 caaggcaggt gaagcaccaa ctgagaaccc tgctccaccc acccagcaga gcagtgctga 1321 ggggccaacc actgaggatc tgtactttca gagcgataac gatggatccg aaatcggtac 1381 tggctttcca ttcgaccccc attatgtgga agtcctgggc gagcgcatgc actacgtcga 1441 tgttggtccg cgcgatggca cccctgtgct gttcctgcac ggtaacccga cctcctccta 1501 cgtgtggcgc aacatcatcc cgcatgttgc accgacccat cgctgcattg ctccagacct 1561 gatcggtatg ggcaaatccg acaaaccaga cctgggttat ttcttcgacg accacgtccg 1621 cttcatggat gccttcatcg aagccctggg tctggaagag gtcgtcctgg tcattcacga 1681 ctggggctcc gctctgggtt tccactgggc caagcgcaat ccagagcgcg tcaaaggtat 1741 tgcatttatg gagttcatcc gccctatccc gacctgggac gaatggccag aatttgcccg 1801 cgagaccttc caggccttcc gcaccaccga cgtcggccgc aagctgatca tcgatcagaa 1861 cgtttttatc gagggtacgc tgccgatggg tgtcgtccgc ccgctgactg aagtcgagat 1921 ggaccattac cgcgagccgt tcctgaatcc tgttgaccgc gagccactgt ggcgcttccc 1981 aaacgagctg ccaatcgccg gtgagccagc gaacatcgtc gcgctggtcg aagaatacat 2041 ggactggctg caccagtccc ctgtcccgaa gctgctgttc tggggcaccc caggcgttct 2101 gatcccaccg gccgaagccg ctcgcctggc caaaagcctg cctaactgca aggctgtgga 2161 catcggcccg ggtctgaatc tgctgcaaga agacaacccg gacctgatcg gcagcgagat 2221 cgcgcgctgg ctgtctactc tggagatttc cggtcatcat caccatcacc actaatagtc 2281 aaccgctgag caataactag cataacccct tggggcctct aaacgggtct tgaggggttt 2341 tttgctgaaa ggaggaacta tatccggttc gcttgctgtc cataaaaccg cccagtctag 2401 ctatcgccat gtaagcccac tgcaagctac ctgctttctc tttgcgcttg cgttttccct 2461 tgtccagata gcccagtagc tgacattcat ccggggtcag caccgtttct gcggactggc 2521 tttctacgta atggtttctt agacgtcagg tggcactttt cggggaaatg tgcgcggaac 2581 ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc 2641 ctgataaatg cttcaataat attgaaaaag gaagagtatg agtattcaac atttccgtgt 2701 cgcccttatt cccttttttg cggcattttg ccttcctgtt tttgctcacc cagaaacgct 2761 ggtgaaagta aaagatgctg aagatcagtt gggtgcacga gtgggttaca tcgaactgga 2821 tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc caatgatgag 2881 cactttcaaa gttctgctat gtggcgcggt attatcccgt attgacgccg ggcaagagca 2941 actcggtcgc cgcatacact attctcagaa tgacttggtt gagtactcac cagtcacaga 3001 aaagcatctt acggatggca tgacagtaag agaattatgc agtgctgcca taaccatgag 3061 tgataacact gcggccaact tacttctgac aactatcgga ggaccgaagg agctaaccgc 3121 ttttttgcac aacatggggg atcatgtaac tcgccttgat cgttgggaac cggagctgaa 3181 tgaagccata ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg caacaacgtt 3241 gcgcaaacta ttaactggcg aactacttac tctagcttcc cggcaacaat taatagactg 3301 gatggaggcg gataaagttg caggaccact tctgcgctcg gcccttccgg ctggctggtt 3361 tattgctgat aaatctggag ccggtgagcg tgggtctcgc ggtatcattg cagcactggg 3421 gccagatggt aagccctccc gtatcgtagt tatctacacg acggggagtc aggcaactat 3481 ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc attggtaatt 3541 cgaaatgacc gaccaagcga cgcccaaccg gtatcagctc actcaaaggc ggtaatacgg 3601 ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag 3661 gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac 3721 gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga 3781 taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt 3841 accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc 3901 tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc 3961 cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta 4021 agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat 4081 gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac tagaaggaca 4141 gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct 4201 tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt 4261 acgcgcagaa aaaaaggatt tcaagaagat cctttgatct tttctacggg gtctgacgct 4321 cagtggaacg aaaactcacg ttaaggg //