LOCUS       sequence-345494-        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_ggHUR-Halo
FEATURES             Location/Qualifiers
     source          1..4460
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     103..122
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        103..121
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      179..188
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1208..1228
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="EDLYFQS"
     CDS             1244..2128
                     /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             2129..2146
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(2157..2175)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      2171..2218
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        2447..2551
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2552..2620,2621..3412)
                     /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(2770..2789)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      complement(3570..4158)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3650..3669)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      4276..4362
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      join(4454..4460,1..21)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
ORIGIN
        1 catcctgacg gatggccttt ttgcgtttct acaaactctt ttgtttattt ttctaaatac
       61 attcaaatat gtatccgctc atggatctcg atcccgcgaa attaatacga ctcactatag
      121 ggtattttaa ctttactaag gagcgatcgc ttccgaattc ctaccgcgga tatctagagc
      181 caccatggcg tctaatggtt atgaagacca catggctgaa gattgcagag atgacatcgg
      241 cagaacaaat ctaattgtga actacctccc ccagaacatg acgcaggatg agctacggag
      301 cctatttagc agtattggcg aagtggaatc tgcaaagctg attcgggaca aagttgcagg
      361 acacagctta ggctatggtt ttgtgaacta cgttactgca aaagatgcag aaagagcaat
      421 aaacacactg aacggcctca gactgcagtc aaaaacgatc aaggtttcct atgctcgtcc
      481 aagttctgaa gttatcaagg acgctaattt atacatcagt gggctcccga ggtcgatgac
      541 acagaaggat gtagaggata tgttctcacg ttttgggcgc atcatcaatt cccgtgtgct
      601 cgtggatcaa accacaggtt tatccagagg ggtcgcattt atccggtttg acaaaaggtc
      661 agaagcagaa gaggcaatta caaatttcaa tggtcacaaa cccccaggtt cctccgaacc
      721 catcacggtg aagtttgcag ccaatcccaa ccagaacaaa aacgtggccc tgttgtcgca
      781 gctgtgccat tcaccagcga ggcggttcgg agggccggtc caccaccagg cgcagagatt
      841 caggttctct cctatgggtg tagatcacat gagtgggctt tctggtgtaa atgttccagg
      901 aaacgcatct tctggctggt gtatcttcat ctacaacctt ggtcaagatg ctgatgaggg
      961 aatcctctgg cagatgtttg gcccctttgg tgcggttacc aatgtgaaag ttattcggga
     1021 tttcaacacc aacaagtgca aaggttttgg ttttgtgacc atgacaaact atgaagaagc
     1081 tgcaatggcc atagcaagtc ttaatggcta ccgcctgggg gacaaaatct tacaggtttc
     1141 cttcaaaacc aacaagtccc acaaaggcgg ctccggctcc tgcaggcgag ctctcgagcc
     1201 aaccactgag gatctgtact ttcagagcga taacgatgga tccgaaatcg gtactggctt
     1261 tccattcgac ccccattatg tggaagtcct gggcgagcgc atgcactacg tcgatgttgg
     1321 tccgcgcgat ggcacccctg tgctgttcct gcacggtaac ccgacctcct cctacgtgtg
     1381 gcgcaacatc atcccgcatg ttgcaccgac ccatcgctgc attgctccag acctgatcgg
     1441 tatgggcaaa tccgacaaac cagacctggg ttatttcttc gacgaccacg tccgcttcat
     1501 ggatgccttc atcgaagccc tgggtctgga agaggtcgtc ctggtcattc acgactgggg
     1561 ctccgctctg ggtttccact gggccaagcg caatccagag cgcgtcaaag gtattgcatt
     1621 tatggagttc atccgcccta tcccgacctg ggacgaatgg ccagaatttg cccgcgagac
     1681 cttccaggcc ttccgcacca ccgacgtcgg ccgcaagctg atcatcgatc agaacgtttt
     1741 tatcgagggt acgctgccga tgggtgtcgt ccgcccgctg actgaagtcg agatggacca
     1801 ttaccgcgag ccgttcctga atcctgttga ccgcgagcca ctgtggcgct tcccaaacga
     1861 gctgccaatc gccggtgagc cagcgaacat cgtcgcgctg gtcgaagaat acatggactg
     1921 gctgcaccag tcccctgtcc cgaagctgct gttctggggc accccaggcg ttctgatccc
     1981 accggccgaa gccgctcgcc tggccaaaag cctgcctaac tgcaaggctg tggacatcgg
     2041 cccgggtctg aatctgctgc aagaagacaa cccggacctg atcggcagcg agatcgcgcg
     2101 ctggctgtct actctggaga tttccggtca tcatcaccat caccactaat agtcaaccgc
     2161 tgagcaataa ctagcataac cccttggggc ctctaaacgg gtcttgaggg gttttttgct
     2221 gaaaggagga actatatccg gttcgcttgc tgtccataaa accgcccagt ctagctatcg
     2281 ccatgtaagc ccactgcaag ctacctgctt tctctttgcg cttgcgtttt cccttgtcca
     2341 gatagcccag tagctgacat tcatccgggg tcagcaccgt ttctgcggac tggctttcta
     2401 cgtaatggtt tcttagacgt caggtggcac ttttcgggga aatgtgcgcg gaacccctat
     2461 ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata
     2521 aatgcttcaa taatattgaa aaaggaagag tatgagtatt caacatttcc gtgtcgccct
     2581 tattcccttt tttgcggcat tttgccttcc tgtttttgct cacccagaaa cgctggtgaa
     2641 agtaaaagat gctgaagatc agttgggtgc acgagtgggt tacatcgaac tggatctcaa
     2701 cagcggtaag atccttgaga gttttcgccc cgaagaacgt tttccaatga tgagcacttt
     2761 caaagttctg ctatgtggcg cggtattatc ccgtattgac gccgggcaag agcaactcgg
     2821 tcgccgcata cactattctc agaatgactt ggttgagtac tcaccagtca cagaaaagca
     2881 tcttacggat ggcatgacag taagagaatt atgcagtgct gccataacca tgagtgataa
     2941 cactgcggcc aacttacttc tgacaactat cggaggaccg aaggagctaa ccgctttttt
     3001 gcacaacatg ggggatcatg taactcgcct tgatcgttgg gaaccggagc tgaatgaagc
     3061 cataccaaac gacgagcgtg acaccacgat gcctgtagca atggcaacaa cgttgcgcaa
     3121 actattaact ggcgaactac ttactctagc ttcccggcaa caattaatag actggatgga
     3181 ggcggataaa gttgcaggac cacttctgcg ctcggccctt ccggctggct ggtttattgc
     3241 tgataaatct ggagccggtg agcgtgggtc tcgcggtatc attgcagcac tggggccaga
     3301 tggtaagccc tcccgtatcg tagttatcta cacgacgggg agtcaggcaa ctatggatga
     3361 acgaaataga cagatcgctg agataggtgc ctcactgatt aagcattggt aattcgaaat
     3421 gaccgaccaa gcgacgccca accggtatca gctcactcaa aggcggtaat acggttatcc
     3481 acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg
     3541 aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat
     3601 cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag
     3661 gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga
     3721 tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg
     3781 tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt
     3841 cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac
     3901 gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc
     3961 ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag gacagtattt
     4021 ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc
     4081 ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc
     4141 agaaaaaaag gatttcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg
     4201 aacgaaaact cacgttaagg gcatatgatg gtagtgtggg gtctccccat gcgagagtag
     4261 ggaactgcca ggcatcaaat aaaacgaaag gctcagtcga aagactgggc ctttcgtttt
     4321 atctgttgtt tgtcggtgaa cgctctcctg agtaggacaa atccgccggg agcggatttg
     4381 aacgttgcga agcaacggcc cggagggtgg cgggcaggac gcccgccata aactgccagg
     4441 catcaaatta agcagaaggc
//
