LOCUS       sequence-345493-        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_mdHUR-Halo
FEATURES             Location/Qualifiers
     source          1..4460
                     /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      1119..1128
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1248..1268
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="EDLYFQS"
     CDS             1284..2168
                     /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             2169..2186
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(2197..2215)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      2211..2258
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        2487..2591
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2592..2660,2661..3452)
                     /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(2810..2829)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      complement(3610..4198)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3690..3709)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      4316..4402
                     /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 tctaatggtt
      241 atgaagacca catggctgaa gattgcaggg atgatatcgg tagaacgaat ttaattgtca
      301 actacctccc tcagaatatg acacaggatg aattgcgaag tttgtttagc agtattggtg
      361 aagttgaatc tgcaaaactt attcgtgata aagtagcagg acacagctta ggctatggct
      421 ttgtgaatta cgtgactgca aaagatgccg aaagagcaat aaacacactg aatggcctga
      481 gactccagtc aaaaaccatt aaggtatcct atgctcgacc aagctcagaa gttatcaaag
      541 atgccaattt gtatattagt ggactcccaa ggacaatgac acagaaagat gtggaggaca
      601 tgttttcacg atttggacgc atcatcaact cccgtgtact tgtggatcag acaacaggtt
      661 tgtcgagagg ggttgcattt atccggtttg acaaaagatc agaagcagaa gaggcaatta
      721 ccagtttcaa tggccataaa cccccaggtt cttccgaacc cataacagtg aagtttgcag
      781 ccaatcctaa ccagaataaa aatgtggcgc tgctctctca gctgtatcac tcaccagcta
      841 gacgatttgg agggccagtt catcatcagg cacaaagatt caggttctcc cctatgggtg
      901 tagatcacat gactgggctt tctggtgtaa atgtcccagg aaacgcatct tctggctggt
      961 gcatcttcat ctacaacctt ggtcaagatg ctgatgaggg aatcctctgg cagatgtttg
     1021 gcccctttgg tgcggttacc aatgtgaaag ttattcgcga tttcaacacc aacaagtgca
     1081 aaggttttgg ttttgtgacc atgacaaact atgaagaagc cgccatggcc atagcaagtc
     1141 tcaatggcta ccgcctgggg gacaaaatct tgcaggtttc cttcaaaacc aacaaggccc
     1201 acaaaggcgg ctccggctcc tgcaggcgag ctctcgagcc aaccactgag gatctgtact
     1261 ttcagagcga taacgatgga tccgaaatcg gtactggctt tccattcgac ccccattatg
     1321 tggaagtcct gggcgagcgc atgcactacg tcgatgttgg tccgcgcgat ggcacccctg
     1381 tgctgttcct gcacggtaac ccgacctcct cctacgtgtg gcgcaacatc atcccgcatg
     1441 ttgcaccgac ccatcgctgc attgctccag acctgatcgg tatgggcaaa tccgacaaac
     1501 cagacctggg ttatttcttc gacgaccacg tccgcttcat ggatgccttc atcgaagccc
     1561 tgggtctgga agaggtcgtc ctggtcattc acgactgggg ctccgctctg ggtttccact
     1621 gggccaagcg caatccagag cgcgtcaaag gtattgcatt tatggagttc atccgcccta
     1681 tcccgacctg ggacgaatgg ccagaatttg cccgcgagac cttccaggcc ttccgcacca
     1741 ccgacgtcgg ccgcaagctg atcatcgatc agaacgtttt tatcgagggt acgctgccga
     1801 tgggtgtcgt ccgcccgctg actgaagtcg agatggacca ttaccgcgag ccgttcctga
     1861 atcctgttga ccgcgagcca ctgtggcgct tcccaaacga gctgccaatc gccggtgagc
     1921 cagcgaacat cgtcgcgctg gtcgaagaat acatggactg gctgcaccag tcccctgtcc
     1981 cgaagctgct gttctggggc accccaggcg ttctgatccc accggccgaa gccgctcgcc
     2041 tggccaaaag cctgcctaac tgcaaggctg tggacatcgg cccgggtctg aatctgctgc
     2101 aagaagacaa cccggacctg atcggcagcg agatcgcgcg ctggctgtct actctggaga
     2161 tttccggtca tcatcaccat caccactaat agtcaaccgc tgagcaataa ctagcataac
     2221 cccttggggc ctctaaacgg gtcttgaggg gttttttgct gaaaggagga actatatccg
     2281 gttcgcttgc tgtccataaa accgcccagt ctagctatcg ccatgtaagc ccactgcaag
     2341 ctacctgctt tctctttgcg cttgcgtttt cccttgtcca gatagcccag tagctgacat
     2401 tcatccgggg tcagcaccgt ttctgcggac tggctttcta cgtaatggtt tcttagacgt
     2461 caggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac
     2521 attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa
     2581 aaaggaagag tatgagtatt caacatttcc gtgtcgccct tattcccttt tttgcggcat
     2641 tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat gctgaagatc
     2701 agttgggtgc acgagtgggt tacatcgaac tggatctcaa cagcggtaag atccttgaga
     2761 gttttcgccc cgaagaacgt tttccaatga tgagcacttt caaagttctg ctatgtggcg
     2821 cggtattatc ccgtattgac gccgggcaag agcaactcgg tcgccgcata cactattctc
     2881 agaatgactt ggttgagtac tcaccagtca cagaaaagca tcttacggat ggcatgacag
     2941 taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc aacttacttc
     3001 tgacaactat cggaggaccg aaggagctaa ccgctttttt gcacaacatg ggggatcatg
     3061 taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac gacgagcgtg
     3121 acaccacgat gcctgtagca atggcaacaa cgttgcgcaa actattaact ggcgaactac
     3181 ttactctagc ttcccggcaa caattaatag actggatgga ggcggataaa gttgcaggac
     3241 cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct ggagccggtg
     3301 agcgtgggtc tcgcggtatc attgcagcac tggggccaga tggtaagccc tcccgtatcg
     3361 tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga cagatcgctg
     3421 agataggtgc ctcactgatt aagcattggt aattcgaaat gaccgaccaa gcgacgccca
     3481 accggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc
     3541 aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt
     3601 gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag
     3661 tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc
     3721 cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc
     3781 ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt
     3841 cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt
     3901 atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc
     3961 agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa
     4021 gtggtggcct aactacggct acactagaag gacagtattt ggtatctgcg ctctgctgaa
     4081 gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg
     4141 tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatttcaaga
     4201 agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg
     4261 gcatatgatg gtagtgtggg gtctccccat gcgagagtag ggaactgcca ggcatcaaat
     4321 aaaacgaaag gctcagtcga aagactgggc ctttcgtttt atctgttgtt tgtcggtgaa
     4381 cgctctcctg agtaggacaa atccgccggg agcggatttg aacgttgcga agcaacggcc
     4441 cggagggtgg cgggcaggac
//
