LOCUS       sequence-345608-        4454 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 4454)
  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 4454)
  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_drHUR-Halo
FEATURES             Location/Qualifiers
     source          1..4454
                     /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"
     regulatory      219..228
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     regulatory      1113..1122
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1242..1262
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="EDLYFQS"
     CDS             1278..2162
                     /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             2163..2180
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(2191..2209)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      2205..2252
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        2481..2585
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2586..2654,2655..3446)
                     /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(2804..2823)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      complement(3604..4192)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3684..3703)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      4310..4396
                     /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 tatctagagc caccatggcg tcgaacggtt
      241 acgaagacca catggccgat gagctcatcg actccaaaac caaccttatc gtcaactacc
      301 tgccacagaa tatgagccag gatgagctgc ggagtctctt cagcagcatt ggggaggtgg
      361 agtctgccaa acttatccga gacaaagtag caggccacag tttagggtac ggatttgtta
      421 actatcttaa ccctaatgat gcagaaagag caatcagtac tctcaatgga ctgagactac
      481 agtctaaaac tatcaaggtg tcatatgcca ggccaagctc tgactccatc aaggatgcta
      541 atctttacat cagtgggctg cctaaaacaa tgacacaaaa ggatgttgaa gaaatgttcg
      601 gacgttatgg tcgaataatc aactcccgtg tacttgttga tcaggcatca ggactctctc
      661 gtggtgtggc tttcattcga tttgacaaaa gggcagaggc agaggacgca atcaaggact
      721 tgaatggaca aaaaccacca ggcgctgctg agcagatgac tgtgaagttt gcagccagtc
      781 ccaaccaagt gaaaaacaca caagttattc cccaggtgta tcaccaacag tctcgccgct
      841 ttggaggacc cgtccaccac caggcccaga gattcaggtt ttctccgatg agcgtggacc
      901 acatgagcgg catgtctggt gtaaacgtgc ctggaaactc ttcatcaggc tggtgcatct
      961 tcatctacaa cttgggccag gacgcagatg agggcattct gtggcagatg ttcgggccct
     1021 tcggcgcagt cacaaacgtc aaagttatcc gtgacttcaa caccaacaag tgcaaaggat
     1081 ttgggtttgt taccatgaca cactacgaag aggccgccat ggctatcgcc agtctcaacg
     1141 gctaccgtct cggggacaag attttacaag tgtcattcaa aacaagcaag tcgcacaagg
     1201 gcggctccgg ctcctgcagg cgagctctcg agccaaccac tgaggatctg tactttcaga
     1261 gcgataacga tggatccgaa atcggtactg gctttccatt cgacccccat tatgtggaag
     1321 tcctgggcga gcgcatgcac tacgtcgatg ttggtccgcg cgatggcacc cctgtgctgt
     1381 tcctgcacgg taacccgacc tcctcctacg tgtggcgcaa catcatcccg catgttgcac
     1441 cgacccatcg ctgcattgct ccagacctga tcggtatggg caaatccgac aaaccagacc
     1501 tgggttattt cttcgacgac cacgtccgct tcatggatgc cttcatcgaa gccctgggtc
     1561 tggaagaggt cgtcctggtc attcacgact ggggctccgc tctgggtttc cactgggcca
     1621 agcgcaatcc agagcgcgtc aaaggtattg catttatgga gttcatccgc cctatcccga
     1681 cctgggacga atggccagaa tttgcccgcg agaccttcca ggccttccgc accaccgacg
     1741 tcggccgcaa gctgatcatc gatcagaacg tttttatcga gggtacgctg ccgatgggtg
     1801 tcgtccgccc gctgactgaa gtcgagatgg accattaccg cgagccgttc ctgaatcctg
     1861 ttgaccgcga gccactgtgg cgcttcccaa acgagctgcc aatcgccggt gagccagcga
     1921 acatcgtcgc gctggtcgaa gaatacatgg actggctgca ccagtcccct gtcccgaagc
     1981 tgctgttctg gggcacccca ggcgttctga tcccaccggc cgaagccgct cgcctggcca
     2041 aaagcctgcc taactgcaag gctgtggaca tcggcccggg tctgaatctg ctgcaagaag
     2101 acaacccgga cctgatcggc agcgagatcg cgcgctggct gtctactctg gagatttccg
     2161 gtcatcatca ccatcaccac taatagtcaa ccgctgagca ataactagca taaccccttg
     2221 gggcctctaa acgggtcttg aggggttttt tgctgaaagg aggaactata tccggttcgc
     2281 ttgctgtcca taaaaccgcc cagtctagct atcgccatgt aagcccactg caagctacct
     2341 gctttctctt tgcgcttgcg ttttcccttg tccagatagc ccagtagctg acattcatcc
     2401 ggggtcagca ccgtttctgc ggactggctt tctacgtaat ggtttcttag acgtcaggtg
     2461 gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa
     2521 atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga
     2581 agagtatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc
     2641 ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg
     2701 gtgcacgagt gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc
     2761 gccccgaaga acgttttcca atgatgagca ctttcaaagt tctgctatgt ggcgcggtat
     2821 tatcccgtat tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg
     2881 acttggttga gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag
     2941 aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa
     3001 ctatcggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc
     3061 gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca
     3121 cgatgcctgt agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc
     3181 tagcttcccg gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc
     3241 tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg
     3301 ggtctcgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta
     3361 tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag
     3421 gtgcctcact gattaagcat tggtaattcg aaatgaccga ccaagcgacg cccaaccggt
     3481 atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa
     3541 gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc
     3601 gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag
     3661 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt
     3721 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg
     3781 aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg
     3841 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg
     3901 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac
     3961 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg
     4021 gcctaactac ggctacacta gaaggacagt atttggtatc tgcgctctgc tgaagccagt
     4081 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg
     4141 tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatttc aagaagatcc
     4201 tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggcatat
     4261 gatggtagtg tggggtctcc ccatgcgaga gtagggaact gccaggcatc aaataaaacg
     4321 aaaggctcag tcgaaagact gggcctttcg ttttatctgt tgtttgtcgg tgaacgctct
     4381 cctgagtagg acaaatccgc cgggagcgga tttgaacgtt gcgaagcaac ggcccggagg
     4441 gtggcgggca ggac
//
