LOCUS       sequence-345488-        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_hsHUR-Halo
FEATURES             Location/Qualifiers
     source          1..4460
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      27..54
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     primer_bind     136..155
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        136..154
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      212..221
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1241..1261
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="EDLYFQS"
     CDS             1277..2161
                     /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             2162..2179
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(2190..2208)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      2204..2251
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        2480..2584
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2585..2653,2654..3445)
                     /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(2803..2822)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      complement(3603..4191)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3683..3702)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      4309..4395
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
ORIGIN
        1 ataaactgcc aggcatcaaa ttaagcagaa ggccatcctg acggatggcc tttttgcgtt
       61 tctacaaact cttttgttta tttttctaaa tacattcaaa tatgtatccg ctcatggatc
      121 tcgatcccgc gaaattaata cgactcacta tagggtattt taactttact aaggagcgat
      181 cgcttccgaa ttcctaccgc ggatatctag agccaccatg gcgtctaatg gttatgaaga
      241 ccacatggcc gaagactgca ggggtgacat cgggagaacg aatttgatcg tcaactacct
      301 ccctcagaac atgacccagg atgagttacg aagcctgttc agcagcattg gtgaagttga
      361 atctgcaaaa cttattcggg ataaagtagc aggacacagc ttgggctatg gctttgtgaa
      421 ctacgtgacc gcgaaggatg cagagagagc gatcaacacg ctgaacggct tgaggctcca
      481 gtcaaaaacc attaaggtgt cgtatgctcg cccgagctca gaggtgatca aagacgccaa
      541 cttgtacatc agcgggctcc cgcggaccat gacccagaag gacgtagaag acatgttctc
      601 tcggtttggg cggatcatca actcgcgggt cctcgtggat cagactacag gtttgtccag
      661 aggggttgcg tttatccggt ttgacaaacg gtcggaggca gaagaggcaa ttaccagttt
      721 caatggtcat aaacccccag gttcctctga gcccatcaca gtgaagtttg cagccaaccc
      781 caaccagaac aaaaacgtgg cactcctctc gcagctgtac cactcgccag cgcgacggtt
      841 cggaggcccc gttcaccacc aggcgcagag attcaggttc tcccccatgg gcgtcgatca
      901 catgagcggg ctctctggcg tcaacgtgcc aggaaacgcc tcctccggct ggtgcatttt
      961 catctacaac ctggggcagg atgccgacga ggggatcctc tggcagatgt ttgggccgtt
     1021 tggtgccgtc accaatgtga aagtgatccg cgacttcaac accaacaagt gcaaagggtt
     1081 tggctttgtg accatgacaa actatgaaga agccgcgatg gccatagcca gcctgaacgg
     1141 ctaccgcctg ggggacaaaa tcttacaggt ttccttcaaa accaacaagt cccacaaagg
     1201 cggctccggc tcctgcaggc gagctctcga gccaaccact gaggatctgt actttcagag
     1261 cgataacgat ggatccgaaa tcggtactgg ctttccattc gacccccatt atgtggaagt
     1321 cctgggcgag cgcatgcact acgtcgatgt tggtccgcgc gatggcaccc ctgtgctgtt
     1381 cctgcacggt aacccgacct cctcctacgt gtggcgcaac atcatcccgc atgttgcacc
     1441 gacccatcgc tgcattgctc cagacctgat cggtatgggc aaatccgaca aaccagacct
     1501 gggttatttc ttcgacgacc acgtccgctt catggatgcc ttcatcgaag ccctgggtct
     1561 ggaagaggtc gtcctggtca ttcacgactg gggctccgct ctgggtttcc actgggccaa
     1621 gcgcaatcca gagcgcgtca aaggtattgc atttatggag ttcatccgcc ctatcccgac
     1681 ctgggacgaa tggccagaat ttgcccgcga gaccttccag gccttccgca ccaccgacgt
     1741 cggccgcaag ctgatcatcg atcagaacgt ttttatcgag ggtacgctgc cgatgggtgt
     1801 cgtccgcccg ctgactgaag tcgagatgga ccattaccgc gagccgttcc tgaatcctgt
     1861 tgaccgcgag ccactgtggc gcttcccaaa cgagctgcca atcgccggtg agccagcgaa
     1921 catcgtcgcg ctggtcgaag aatacatgga ctggctgcac cagtcccctg tcccgaagct
     1981 gctgttctgg ggcaccccag gcgttctgat cccaccggcc gaagccgctc gcctggccaa
     2041 aagcctgcct aactgcaagg ctgtggacat cggcccgggt ctgaatctgc tgcaagaaga
     2101 caacccggac ctgatcggca gcgagatcgc gcgctggctg tctactctgg agatttccgg
     2161 tcatcatcac catcaccact aatagtcaac cgctgagcaa taactagcat aaccccttgg
     2221 ggcctctaaa cgggtcttga ggggtttttt gctgaaagga ggaactatat ccggttcgct
     2281 tgctgtccat aaaaccgccc agtctagcta tcgccatgta agcccactgc aagctacctg
     2341 ctttctcttt gcgcttgcgt tttcccttgt ccagatagcc cagtagctga cattcatccg
     2401 gggtcagcac cgtttctgcg gactggcttt ctacgtaatg gtttcttaga cgtcaggtgg
     2461 cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa
     2521 tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa
     2581 gagtatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct
     2641 tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg
     2701 tgcacgagtg ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg
     2761 ccccgaagaa cgttttccaa tgatgagcac tttcaaagtt ctgctatgtg gcgcggtatt
     2821 atcccgtatt gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga
     2881 cttggttgag tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga
     2941 attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac
     3001 tatcggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg
     3061 ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac
     3121 gatgcctgta gcaatggcaa caacgttgcg caaactatta actggcgaac tacttactct
     3181 agcttcccgg caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct
     3241 gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg
     3301 gtctcgcggt atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat
     3361 ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg
     3421 tgcctcactg attaagcatt ggtaattcga aatgaccgac caagcgacgc ccaaccggta
     3481 tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag
     3541 aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg
     3601 tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg
     3661 tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg
     3721 cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga
     3781 agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc
     3841 tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt
     3901 aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact
     3961 ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg
     4021 cctaactacg gctacactag aaggacagta tttggtatct gcgctctgct gaagccagtt
     4081 accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt
     4141 ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatttca agaagatcct
     4201 ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggcatatg
     4261 atggtagtgt ggggtctccc catgcgagag tagggaactg ccaggcatca aataaaacga
     4321 aaggctcagt cgaaagactg ggcctttcgt tttatctgtt gtttgtcggt gaacgctctc
     4381 ctgagtagga caaatccgcc gggagcggat ttgaacgttg cgaagcaacg gcccggaggg
     4441 tggcgggcag gacgcccgcc
//
