LOCUS       sequence-344834-        4317 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 4317)
  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 4317)
  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_HNRNPC-Halo
FEATURES             Location/Qualifiers
     source          1..4317
                     /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"
     CDS             1304..1324
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="EDLYFQS"
     CDS             1340..2224
                     /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             2225..2242
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(2253..2271)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      2267..2314
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        2543..2647
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2648..2716,2717..3508)
                     /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(2866..2885)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      complement(3666..4254)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3746..3765)
                     /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 atatggccag
      421 caacgttacc aacaagacag atcctcgctc catgaactcc cgtgtattca ttgggaatct
      481 caacactctt gtggtcaaga aatctgatgt ggaggcaatc ttttcgaagt atggcaaaat
      541 tgtgggctgc tctgttcata agggctttgc cttcgttcag tatgttaatg agagaaatgc
      601 ccgggctgct gtagcaggag aggatggcag aatgattgct ggccaggttt tagatattaa
      661 cctggctgca gagccaaaag tgaaccgagg aaaagcaggt gtgaaacgat ctgcagcgga
      721 gatgtacggc tcctcttttg acttggacta tgactttcaa cgggactatt atgataggat
      781 gtacagttac ccagcacgtg tacctcctcc tcctcctatt gctcgggctg tagtgccctc
      841 gaaacgtcag cgtgtatcag gaaacacttc acgaaggggc aaaagtggct tcaattctaa
      901 gagtggacag cggggatctt ccaagtctgg aaagttgaaa ggagatgacc ttcaggccat
      961 taagaaggag ctgacccaga taaaacaaaa agtggattct ctcctggaaa acctggaaaa
     1021 aattgaaaag gaacagagca aacaagcagt agagatgaag aatgataagt cagaagagga
     1081 gcagagcagc agctccgtga agaaagatga gactaatgtg aagatggagt ctgagggggg
     1141 tgcagatgac tctgctgagg agggggacct actggatgat gatgataatg aagatcgggg
     1201 ggatgaccag ctggagttga tcaaggatga tgaaaaagag gctgaggaag gagaggatga
     1261 cagagacagc gccaatggcg aggatgactc tgggccaacc actgaggatc tgtactttca
     1321 gagcgataac gatggatccg aaatcggtac tggctttcca ttcgaccccc attatgtgga
     1381 agtcctgggc gagcgcatgc actacgtcga tgttggtccg cgcgatggca cccctgtgct
     1441 gttcctgcac ggtaacccga cctcctccta cgtgtggcgc aacatcatcc cgcatgttgc
     1501 accgacccat cgctgcattg ctccagacct gatcggtatg ggcaaatccg acaaaccaga
     1561 cctgggttat ttcttcgacg accacgtccg cttcatggat gccttcatcg aagccctggg
     1621 tctggaagag gtcgtcctgg tcattcacga ctggggctcc gctctgggtt tccactgggc
     1681 caagcgcaat ccagagcgcg tcaaaggtat tgcatttatg gagttcatcc gccctatccc
     1741 gacctgggac gaatggccag aatttgcccg cgagaccttc caggccttcc gcaccaccga
     1801 cgtcggccgc aagctgatca tcgatcagaa cgtttttatc gagggtacgc tgccgatggg
     1861 tgtcgtccgc ccgctgactg aagtcgagat ggaccattac cgcgagccgt tcctgaatcc
     1921 tgttgaccgc gagccactgt ggcgcttccc aaacgagctg ccaatcgccg gtgagccagc
     1981 gaacatcgtc gcgctggtcg aagaatacat ggactggctg caccagtccc ctgtcccgaa
     2041 gctgctgttc tggggcaccc caggcgttct gatcccaccg gccgaagccg ctcgcctggc
     2101 caaaagcctg cctaactgca aggctgtgga catcggcccg ggtctgaatc tgctgcaaga
     2161 agacaacccg gacctgatcg gcagcgagat cgcgcgctgg ctgtctactc tggagatttc
     2221 cggtcatcat caccatcacc actaatagtc aaccgctgag caataactag cataacccct
     2281 tggggcctct aaacgggtct tgaggggttt tttgctgaaa ggaggaacta tatccggttc
     2341 gcttgctgtc cataaaaccg cccagtctag ctatcgccat gtaagcccac tgcaagctac
     2401 ctgctttctc tttgcgcttg cgttttccct tgtccagata gcccagtagc tgacattcat
     2461 ccggggtcag caccgtttct gcggactggc tttctacgta atggtttctt agacgtcagg
     2521 tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc
     2581 aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag
     2641 gaagagtatg agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg
     2701 ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta aaagatgctg aagatcagtt
     2761 gggtgcacga gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt
     2821 tcgccccgaa gaacgttttc caatgatgag cactttcaaa gttctgctat gtggcgcggt
     2881 attatcccgt attgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa
     2941 tgacttggtt gagtactcac cagtcacaga aaagcatctt acggatggca tgacagtaag
     3001 agaattatgc agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac
     3061 aactatcgga ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac
     3121 tcgccttgat cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac
     3181 cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg aactacttac
     3241 tctagcttcc cggcaacaat taatagactg gatggaggcg gataaagttg caggaccact
     3301 tctgcgctcg gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg
     3361 tgggtctcgc ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt
     3421 tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat
     3481 aggtgcctca ctgattaagc attggtaatt cgaaatgacc gaccaagcga cgcccaaccg
     3541 gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga
     3601 aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg
     3661 gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag
     3721 aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc
     3781 gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg
     3841 ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt
     3901 cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc
     3961 ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc
     4021 actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg
     4081 tggcctaact acggctacac tagaaggaca gtatttggta tctgcgctct gctgaagcca
     4141 gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc
     4201 ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatt tcaagaagat
     4261 cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaaggg
//
