LOCUS       sequence-345602-        4086 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Production of HaloTag fusion protein.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4086)
  AUTHORS   Atanasoai I, Papavasileiou S, Preiß N, Kutter C
  TITLE     Large-scale identification of RBP-RNA interactions by RAPseq refines
            essentials of post-transcriptional gene regulation
  JOURNAL   bioRxiv 2021.11.08.467743
REFERENCE   2  (bases 1 to 4086)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "bioRxiv 
            2021.11.08.467743"
COMMENT     Sequence Label: pH6HTC_RAN-Halo
FEATURES             Location/Qualifiers
     source          1..4086
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      34..61
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     primer_bind     143..162
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        143..161
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             874..894
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="EDLYFQS"
     CDS             910..1794
                     /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             1795..1812
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(1823..1841)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1837..1884
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        2113..2217
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2218..2286,2287..3078)
                     /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(2436..2455)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      complement(3236..3824)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3316..3335)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      3942..4028
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
ORIGIN
        1 gcccgccata aactgccagg catcaaatta agcagaaggc catcctgacg gatggccttt
       61 ttgcgtttct acaaactctt ttgtttattt ttctaaatac attcaaatat gtatccgctc
      121 atggatctcg atcccgcgaa attaatacga ctcactatag ggtattttaa ctttactaag
      181 gagcgatcgc ttccgaattc ctaccgcgga tatatggctg cgcagggaga gccccaggtc
      241 cagttcaaac ttgtattggt tggtgatggt ggtactggaa aaacgacctt cgtgaaacgt
      301 catttgactg gtgaatttga gaagaagtat gtagccacct tgggtgttga ggttcatccc
      361 ctagtgttcc acaccaacag aggacctatt aagttcaatg tatgggacac agccggccag
      421 gagaaattcg gtggactgag agatggctat tatatccaag cccagtgtgc catcataatg
      481 tttgatgtaa catcgagagt tacttacaag aatgtgccta actggcatag agatctggta
      541 cgagtgtgtg aaaacatccc cattgtgttg tgtggcaaca aagtggatat taaggacagg
      601 aaagtgaagg cgaaatccat tgtcttccac cgaaagaaga atcttcagta ctacgacatt
      661 tctgccaaaa gtaactacaa ctttgaaaag cccttcctct ggcttgctag gaagctcatt
      721 ggagacccta acttggaatt tgttgccatg cctgctctcg ccccaccaga agttgtcatg
      781 gacccagctt tggcagcaca gtatgagcac gacttagagg ttgctcagac aactgctctc
      841 ccggatgagg atgatgacct ggagccaacc actgaggatc tgtactttca gagcgataac
      901 gatggatccg aaatcggtac tggctttcca ttcgaccccc attatgtgga agtcctgggc
      961 gagcgcatgc actacgtcga tgttggtccg cgcgatggca cccctgtgct gttcctgcac
     1021 ggtaacccga cctcctccta cgtgtggcgc aacatcatcc cgcatgttgc accgacccat
     1081 cgctgcattg ctccagacct gatcggtatg ggcaaatccg acaaaccaga cctgggttat
     1141 ttcttcgacg accacgtccg cttcatggat gccttcatcg aagccctggg tctggaagag
     1201 gtcgtcctgg tcattcacga ctggggctcc gctctgggtt tccactgggc caagcgcaat
     1261 ccagagcgcg tcaaaggtat tgcatttatg gagttcatcc gccctatccc gacctgggac
     1321 gaatggccag aatttgcccg cgagaccttc caggccttcc gcaccaccga cgtcggccgc
     1381 aagctgatca tcgatcagaa cgtttttatc gagggtacgc tgccgatggg tgtcgtccgc
     1441 ccgctgactg aagtcgagat ggaccattac cgcgagccgt tcctgaatcc tgttgaccgc
     1501 gagccactgt ggcgcttccc aaacgagctg ccaatcgccg gtgagccagc gaacatcgtc
     1561 gcgctggtcg aagaatacat ggactggctg caccagtccc ctgtcccgaa gctgctgttc
     1621 tggggcaccc caggcgttct gatcccaccg gccgaagccg ctcgcctggc caaaagcctg
     1681 cctaactgca aggctgtgga catcggcccg ggtctgaatc tgctgcaaga agacaacccg
     1741 gacctgatcg gcagcgagat cgcgcgctgg ctgtctactc tggagatttc cggtcatcat
     1801 caccatcacc actaatagtc aaccgctgag caataactag cataacccct tggggcctct
     1861 aaacgggtct tgaggggttt tttgctgaaa ggaggaacta tatccggttc gcttgctgtc
     1921 cataaaaccg cccagtctag ctatcgccat gtaagcccac tgcaagctac ctgctttctc
     1981 tttgcgcttg cgttttccct tgtccagata gcccagtagc tgacattcat ccggggtcag
     2041 caccgtttct gcggactggc tttctacgta atggtttctt agacgtcagg tggcactttt
     2101 cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat
     2161 ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtatg
     2221 agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt
     2281 tttgctcacc cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcacga
     2341 gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa
     2401 gaacgttttc caatgatgag cactttcaaa gttctgctat gtggcgcggt attatcccgt
     2461 attgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt
     2521 gagtactcac cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc
     2581 agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac aactatcgga
     2641 ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat
     2701 cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct
     2761 gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc
     2821 cggcaacaat taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg
     2881 gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc
     2941 ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg
     3001 acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca
     3061 ctgattaagc attggtaatt cgaaatgacc gaccaagcga cgcccaaccg gtatcagctc
     3121 actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt
     3181 gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc
     3241 ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa
     3301 acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc
     3361 ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg
     3421 cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc
     3481 tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc
     3541 gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca
     3601 ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact
     3661 acggctacac tagaaggaca gtatttggta tctgcgctct gctgaagcca gttaccttcg
     3721 gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt
     3781 ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatt tcaagaagat cctttgatct
     3841 tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggcat atgatggtag
     3901 tgtggggtct ccccatgcga gagtagggaa ctgccaggca tcaaataaaa cgaaaggctc
     3961 agtcgaaaga ctgggccttt cgttttatct gttgtttgtc ggtgaacgct ctcctgagta
     4021 ggacaaatcc gccgggagcg gatttgaacg ttgcgaagca acggcccgga gggtggcggg
     4081 caggac
//
