LOCUS       sequence-344844-        5159 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 5159)
  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 5159)
  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_YTHDF1-Halo
FEATURES             Location/Qualifiers
     source          1..5159
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      55..141
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      233..260
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     primer_bind     342..361
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        342..360
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      418..427
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2146..2166
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="EDLYFQS"
     CDS             2182..3066
                     /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             3067..3084
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(3095..3113)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      3109..3156
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        3385..3489
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3490..3558,3559..4350)
                     /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(3708..3727)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      complement(4508..5096)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4588..4607)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 catatgatgg tagtgtgggg tctccccatg cgagagtagg gaactgccag gcatcaaata
       61 aaacgaaagg ctcagtcgaa agactgggcc tttcgtttta tctgttgttt gtcggtgaac
      121 gctctcctga gtaggacaaa tccgccggga gcggatttga acgttgcgaa gcaacggccc
      181 ggagggtggc gggcaggacg cccgccataa actgccaggc atcaaattaa gcagaaggcc
      241 atcctgacgg atggcctttt tgcgtttcta caaactcttt tgtttatttt tctaaataca
      301 ttcaaatatg tatccgctca tggatctcga tcccgcgaaa ttaatacgac tcactatagg
      361 gtattttaac tttactaagg agcgatcgct tccgaattcc taccgcggat atctagagcc
      421 accatggcgt cggccaccag cgtggacacc cagagaacaa aaggacaaga taataaagta
      481 caaaatggtt cgttacatca gaaggataca gttcatgaca atgactttga gccctacctt
      541 actggacagt caaatcagag taacagttac ccctcaatga gcgaccccta cctgtccagc
      601 tattacccgc cgtccattgg atttccttac tccctcaatg aggctccgtg gtctactgca
      661 ggggaccctc cgattccata cctcaccacc tacggacagc tcagtaacgg agaccatcat
      721 tttatgcacg atgctgtttt tgggcagcct gggggcctgg ggaacaacat ctatcagcac
      781 aggttcaatt ttttccctga aaaccctgcg ttctcagcat gggggacaag tgggtctcaa
      841 ggtcagcaga cccagagctc cgcgtatggg agcagctaca cctacccccc gagctccctg
      901 ggtggcacgg tggttgatgg gcagccaggc tttcacagcg acaccctcag caaggccccc
      961 gggatgaaca gcctggagca gggcatggtt ggcctgaaga ttggggacgt cagctcctcc
     1021 gctgtcaaga cggtgggctc tgtcgtcagc agcgtggcac tgactggtgt cctttctggc
     1081 aacggtggga caaatgtgaa catgccagtt tcaaagccga cctcgtgggc tgccattgcc
     1141 agcaagcctg caaaaccaca gcctaaaatg aaaacaaaga gcgggcctgt catggggggt
     1201 gggctgcccc ctccacccat aaagcataac atggacattg gcacctggga taacaagggg
     1261 cctgtgccga aggccccagt cccccagcag gcaccctctc cacaggctgc cccacagccc
     1321 cagcaggtgg ctcagcctct cccagcacag cccccagctt tggctcaacc gcagtatcag
     1381 agccctcagc agccacccca gacccgctgg gttgccccac gcaacagaaa cgcggcgttt
     1441 gggcagagcg gaggggctgg cagcgatagc aactctcctg gaaacgtcca gcctaattct
     1501 gcccccagcg tcgaatccca ccccgtcctt gaaaaactga aggctgctca cagctacaac
     1561 ccgaaagagt ttgagtggaa tctgaaaagc gggcgtgtgt tcatcatcaa gagctactct
     1621 gaggacgaca tccaccgctc cattaagtac tccatctggt gtagcacaga gcacggcaac
     1681 aagcgcctgg acagcgcctt ccgctgcatg agcagcaagg ggcccgtcta cctgctcttc
     1741 agcgtcaatg ggagtgggca tttttgtggg gtggccgaga tgaagtcccc cgtggactac
     1801 ggcaccagtg ccggggtctg gtctcaggac aagtggaagg ggaagtttga tgtccagtgg
     1861 atttttgtta aggatgtacc caataaccag ctccggcaca tcaggctgga gaataacgac
     1921 aacaaaccgg tcacaaactc ccgggacacc caggaggtgc ccttagaaaa agccaagcaa
     1981 gtgctgaaaa ttatcagttc ctacaagcac acaacctcca tcttcgacga ctttgctcac
     2041 tacgagaagc gccaggagga ggaggaggtg gtgcgcaagg aacggcagag tcgaaacaaa
     2101 caaggcggct ccggctcctg caggcgagct ctcgagccaa ccactgagga tctgtacttt
     2161 cagagcgata acgatggatc cgaaatcggt actggctttc cattcgaccc ccattatgtg
     2221 gaagtcctgg gcgagcgcat gcactacgtc gatgttggtc cgcgcgatgg cacccctgtg
     2281 ctgttcctgc acggtaaccc gacctcctcc tacgtgtggc gcaacatcat cccgcatgtt
     2341 gcaccgaccc atcgctgcat tgctccagac ctgatcggta tgggcaaatc cgacaaacca
     2401 gacctgggtt atttcttcga cgaccacgtc cgcttcatgg atgccttcat cgaagccctg
     2461 ggtctggaag aggtcgtcct ggtcattcac gactggggct ccgctctggg tttccactgg
     2521 gccaagcgca atccagagcg cgtcaaaggt attgcattta tggagttcat ccgccctatc
     2581 ccgacctggg acgaatggcc agaatttgcc cgcgagacct tccaggcctt ccgcaccacc
     2641 gacgtcggcc gcaagctgat catcgatcag aacgttttta tcgagggtac gctgccgatg
     2701 ggtgtcgtcc gcccgctgac tgaagtcgag atggaccatt accgcgagcc gttcctgaat
     2761 cctgttgacc gcgagccact gtggcgcttc ccaaacgagc tgccaatcgc cggtgagcca
     2821 gcgaacatcg tcgcgctggt cgaagaatac atggactggc tgcaccagtc ccctgtcccg
     2881 aagctgctgt tctggggcac cccaggcgtt ctgatcccac cggccgaagc cgctcgcctg
     2941 gccaaaagcc tgcctaactg caaggctgtg gacatcggcc cgggtctgaa tctgctgcaa
     3001 gaagacaacc cggacctgat cggcagcgag atcgcgcgct ggctgtctac tctggagatt
     3061 tccggtcatc atcaccatca ccactaatag tcaaccgctg agcaataact agcataaccc
     3121 cttggggcct ctaaacgggt cttgaggggt tttttgctga aaggaggaac tatatccggt
     3181 tcgcttgctg tccataaaac cgcccagtct agctatcgcc atgtaagccc actgcaagct
     3241 acctgctttc tctttgcgct tgcgttttcc cttgtccaga tagcccagta gctgacattc
     3301 atccggggtc agcaccgttt ctgcggactg gctttctacg taatggtttc ttagacgtca
     3361 ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat
     3421 tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa
     3481 aggaagagta tgagtattca acatttccgt gtcgccctta ttcccttttt tgcggcattt
     3541 tgccttcctg tttttgctca cccagaaacg ctggtgaaag taaaagatgc tgaagatcag
     3601 ttgggtgcac gagtgggtta catcgaactg gatctcaaca gcggtaagat ccttgagagt
     3661 tttcgccccg aagaacgttt tccaatgatg agcactttca aagttctgct atgtggcgcg
     3721 gtattatccc gtattgacgc cgggcaagag caactcggtc gccgcataca ctattctcag
     3781 aatgacttgg ttgagtactc accagtcaca gaaaagcatc ttacggatgg catgacagta
     3841 agagaattat gcagtgctgc cataaccatg agtgataaca ctgcggccaa cttacttctg
     3901 acaactatcg gaggaccgaa ggagctaacc gcttttttgc acaacatggg ggatcatgta
     3961 actcgccttg atcgttggga accggagctg aatgaagcca taccaaacga cgagcgtgac
     4021 accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg cgaactactt
     4081 actctagctt cccggcaaca attaatagac tggatggagg cggataaagt tgcaggacca
     4141 cttctgcgct cggcccttcc ggctggctgg tttattgctg ataaatctgg agccggtgag
     4201 cgtgggtctc gcggtatcat tgcagcactg gggccagatg gtaagccctc ccgtatcgta
     4261 gttatctaca cgacggggag tcaggcaact atggatgaac gaaatagaca gatcgctgag
     4321 ataggtgcct cactgattaa gcattggtaa ttcgaaatga ccgaccaagc gacgcccaac
     4381 cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag
     4441 gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc
     4501 tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc
     4561 agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc
     4621 tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt
     4681 cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg
     4741 ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat
     4801 ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag
     4861 ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt
     4921 ggtggcctaa ctacggctac actagaagga cagtatttgg tatctgcgct ctgctgaagc
     4981 cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta
     5041 gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tttcaagaag
     5101 atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggg
//
