LOCUS       sequence-345073-        4460 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 4460)
  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 4460)
  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_mmHUR-Halo
FEATURES             Location/Qualifiers
     source          1..4460
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      48..75
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     primer_bind     157..176
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        157..175
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      233..242
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1262..1282
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="EDLYFQS"
     CDS             1298..2182
                     /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             2183..2200
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(2211..2229)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      2225..2272
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        2501..2605
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2606..2674,2675..3466)
                     /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(2824..2843)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      complement(3624..4212)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3704..3723)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      4330..4416
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
ORIGIN
        1 gtggcgggca ggacgcccgc cataaactgc caggcatcaa attaagcaga aggccatcct
       61 gacggatggc ctttttgcgt ttctacaaac tcttttgttt atttttctaa atacattcaa
      121 atatgtatcc gctcatggat ctcgatcccg cgaaattaat acgactcact atagggtatt
      181 ttaactttac taaggagcga tcgcttccga attcctaccg cggatatcta gagccaccat
      241 ggcgtctaat ggttatgaag accacatggc ggaagactgc agggatgaca ttgggagaac
      301 gaatttaatt gtcaactacc tccctcagaa catgacccaa gaggaactac gaagtctgtt
      361 cagcagcatt ggcgaggttg aatctgcaaa gcttattcgg gataaagtag caggacacag
      421 cttgggctac ggttttgtga actatgtgac tgcaaaagat gcagagagag caatcagcac
      481 actgaacggc ttgagactcc agtccaaaac cattaaggtg tcatatgctc gcccaagctc
      541 agaggtcatc aaagatgcca acttatacat cagtgggctc ccaaggacca tgacacagaa
      601 ggatgtggaa gacatgtttt ctcggtttgg gcgaatcatc aactccaggg tccttgtgga
      661 tcagaccaca ggtttgtcca gaggggttgc ctttatccgg tttgacaaac ggtcagaagc
      721 agaagaggca attaccagtt tcaatggtca taaaccccca ggttcctccg agcccatcac
      781 agtgaagttt gcagccaatc ccaaccagaa caaaaacatg gctctcctct cgcagctgta
      841 ccactcgcct gctaggcggt ttggaggccc tgtacaccac caggcacaga gattcaggtt
      901 ctcccctatg ggtgtagatc acatgagtgg gatttctggt gtcaatgtcc ccggcaatgc
      961 ttcctcgggc tggtgcatct tcatctacaa ccttgggcaa gacgccgatg aggggatcct
     1021 ctggcagatg tttggcccct ttggtgcagt taccaatgtg aaagtgattc gtgatttcaa
     1081 caccaacaag tgcaaagggt ttggttttgt gaccatgaca aactatgaag aagctgcaat
     1141 ggccatagca agtctgaacg gctaccgcct gggggacaaa attttacagg tttccttcaa
     1201 aaccaacaag tcccacaaag gcggctccgg ctcctgcagg cgagctctcg agccaaccac
     1261 tgaggatctg tactttcaga gcgataacga tggatccgaa atcggtactg gctttccatt
     1321 cgacccccat tatgtggaag tcctgggcga gcgcatgcac tacgtcgatg ttggtccgcg
     1381 cgatggcacc cctgtgctgt tcctgcacgg taacccgacc tcctcctacg tgtggcgcaa
     1441 catcatcccg catgttgcac cgacccatcg ctgcattgct ccagacctga tcggtatggg
     1501 caaatccgac aaaccagacc tgggttattt cttcgacgac cacgtccgct tcatggatgc
     1561 cttcatcgaa gccctgggtc tggaagaggt cgtcctggtc attcacgact ggggctccgc
     1621 tctgggtttc cactgggcca agcgcaatcc agagcgcgtc aaaggtattg catttatgga
     1681 gttcatccgc cctatcccga cctgggacga atggccagaa tttgcccgcg agaccttcca
     1741 ggccttccgc accaccgacg tcggccgcaa gctgatcatc gatcagaacg tttttatcga
     1801 gggtacgctg ccgatgggtg tcgtccgccc gctgactgaa gtcgagatgg accattaccg
     1861 cgagccgttc ctgaatcctg ttgaccgcga gccactgtgg cgcttcccaa acgagctgcc
     1921 aatcgccggt gagccagcga acatcgtcgc gctggtcgaa gaatacatgg actggctgca
     1981 ccagtcccct gtcccgaagc tgctgttctg gggcacccca ggcgttctga tcccaccggc
     2041 cgaagccgct cgcctggcca aaagcctgcc taactgcaag gctgtggaca tcggcccggg
     2101 tctgaatctg ctgcaagaag acaacccgga cctgatcggc agcgagatcg cgcgctggct
     2161 gtctactctg gagatttccg gtcatcatca ccatcaccac taatagtcaa ccgctgagca
     2221 ataactagca taaccccttg gggcctctaa acgggtcttg aggggttttt tgctgaaagg
     2281 aggaactata tccggttcgc ttgctgtcca taaaaccgcc cagtctagct atcgccatgt
     2341 aagcccactg caagctacct gctttctctt tgcgcttgcg ttttcccttg tccagatagc
     2401 ccagtagctg acattcatcc ggggtcagca ccgtttctgc ggactggctt tctacgtaat
     2461 ggtttcttag acgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt
     2521 atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct
     2581 tcaataatat tgaaaaagga agagtatgag tattcaacat ttccgtgtcg cccttattcc
     2641 cttttttgcg gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa
     2701 agatgctgaa gatcagttgg gtgcacgagt gggttacatc gaactggatc tcaacagcgg
     2761 taagatcctt gagagttttc gccccgaaga acgttttcca atgatgagca ctttcaaagt
     2821 tctgctatgt ggcgcggtat tatcccgtat tgacgccggg caagagcaac tcggtcgccg
     2881 catacactat tctcagaatg acttggttga gtactcacca gtcacagaaa agcatcttac
     2941 ggatggcatg acagtaagag aattatgcag tgctgccata accatgagtg ataacactgc
     3001 ggccaactta cttctgacaa ctatcggagg accgaaggag ctaaccgctt ttttgcacaa
     3061 catgggggat catgtaactc gccttgatcg ttgggaaccg gagctgaatg aagccatacc
     3121 aaacgacgag cgtgacacca cgatgcctgt agcaatggca acaacgttgc gcaaactatt
     3181 aactggcgaa ctacttactc tagcttcccg gcaacaatta atagactgga tggaggcgga
     3241 taaagttgca ggaccacttc tgcgctcggc ccttccggct ggctggttta ttgctgataa
     3301 atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc cagatggtaa
     3361 gccctcccgt atcgtagtta tctacacgac ggggagtcag gcaactatgg atgaacgaaa
     3421 tagacagatc gctgagatag gtgcctcact gattaagcat tggtaattcg aaatgaccga
     3481 ccaagcgacg cccaaccggt atcagctcac tcaaaggcgg taatacggtt atccacagaa
     3541 tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt
     3601 aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa
     3661 aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt
     3721 ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg
     3781 tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc
     3841 agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc
     3901 gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta
     3961 tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct
     4021 acagagttct tgaagtggtg gcctaactac ggctacacta gaaggacagt atttggtatc
     4081 tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa
     4141 caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa
     4201 aaaggatttc aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa
     4261 aactcacgtt aagggcatat gatggtagtg tggggtctcc ccatgcgaga gtagggaact
     4321 gccaggcatc aaataaaacg aaaggctcag tcgaaagact gggcctttcg ttttatctgt
     4381 tgtttgtcgg tgaacgctct cctgagtagg acaaatccgc cgggagcgga tttgaacgtt
     4441 gcgaagcaac ggcccggagg
//
