LOCUS       sequence-345598-        5043 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 5043)
  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 5043)
  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_CCT2-Halo
FEATURES             Location/Qualifiers
     source          1..5043
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             1614..1634
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="EDLYFQS"
     CDS             1650..2534
                     /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             2535..2552
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(2563..2581)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      2577..2624
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        2853..2957
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2958..3026,3027..3818)
                     /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(3176..3195)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      complement(3976..4564)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4056..4075)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      4682..4768
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      4860..4887
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     primer_bind     4969..4988
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        4969..4987
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
        1 cgtccctttc ccttgcacct gttaacatct ttaaggcagg agctgatgaa gagagagcag
       61 agacagctcg tctgacttct tttattggtg ccatcgccat tggagacttg gtaaagagca
      121 ccttgggacc caaaggcatg gacaaaattc ttctaagcag tggacgagat gcctctctta
      181 tggtaaccaa tgatggtgcc actattctaa aaaacattgg tgttgacaat ccagcagcta
      241 aagttttagt tgatatgtca agggttcaag atgatgaagt tggtgatggc actacctctg
      301 ttaccgtttt agcagcagaa ttattaaggg aagcagaatc tttaattgca aaaaagattc
      361 atccacagac catcatagcg ggttggagag aagccacgaa ggctgcaaga gaggcgctgt
      421 tgagttctgc agttgatcat ggttccgatg aagttaaatt ccgtcaagat ttaatgaata
      481 ttgcgggcac aacattatcc tcaaaacttc ttactcatca caaagaccac tttacaaagt
      541 tagctgtaga agcagttctc agactgaaag gctctggcaa cctggaggca attcatatta
      601 tcaagaagct aggaggaagt ttggcagatt cctatttaga tgaaggcttc ctgttggata
      661 aaaaaattgg agtaaatcaa ccaaaacgaa ttgaaaatgc taaaattctt attgcaaata
      721 ctggtatgga tacagacaaa ataaagatat ttggttcccg ggtaagagtt gactctacag
      781 caaaggttgc agaaatagaa catgcggaaa aggaaaaaat gaaggagaaa gttgaacgta
      841 ttcttaagca tggaataaat tgctttatta acaggcaatt aatttataat tatcctgaac
      901 agctctttgg tgctgctggt gtcatggcta ttgagcatgc agattttgca ggtgtggaac
      961 gcctagctct tgtcacaggt ggtgaaattg cctctacctt tgatcaccca gaactggtga
     1021 agcttggaag ttgcaaactt atcgaggaag tcatgattgg agaagacaaa ctcattcact
     1081 tttctggggt tgcccttggt gaggcttgta ccattgtttt gcgtggtgcc actcaacaaa
     1141 ttttagatga agcagaaaga tcattgcatg atgctctttg tgttcttgcg caaactgtaa
     1201 aggactctag aacagtttat ggaggaggct gttctgagat gttgatggct catgctgtga
     1261 cacagcttgc caatagaaca ccaggcaaag aagctgttgc aatggagtct tatgctaaag
     1321 cactgagaat gttgccaacc atcatagctg acaatgcagg ctatgacagt gcagacctgg
     1381 tggcacagct cagggctgct cacagtgaag gcaataccac tgctggattg gatatgaggg
     1441 aaggcaccat tggagatatg gctatcctgg gtataacaga aagttttcaa gtgaagcgac
     1501 aggttcttct gagtgcagct gaagcagcag aggtgattct gcgtgtggac aacatcatca
     1561 aagcggcacc caggaaacgt gtccctgatc accacccctg tgggccaacc actgaggatc
     1621 tgtactttca gagcgataac gatggatccg aaatcggtac tggctttcca ttcgaccccc
     1681 attatgtgga agtcctgggc gagcgcatgc actacgtcga tgttggtccg cgcgatggca
     1741 cccctgtgct gttcctgcac ggtaacccga cctcctccta cgtgtggcgc aacatcatcc
     1801 cgcatgttgc accgacccat cgctgcattg ctccagacct gatcggtatg ggcaaatccg
     1861 acaaaccaga cctgggttat ttcttcgacg accacgtccg cttcatggat gccttcatcg
     1921 aagccctggg tctggaagag gtcgtcctgg tcattcacga ctggggctcc gctctgggtt
     1981 tccactgggc caagcgcaat ccagagcgcg tcaaaggtat tgcatttatg gagttcatcc
     2041 gccctatccc gacctgggac gaatggccag aatttgcccg cgagaccttc caggccttcc
     2101 gcaccaccga cgtcggccgc aagctgatca tcgatcagaa cgtttttatc gagggtacgc
     2161 tgccgatggg tgtcgtccgc ccgctgactg aagtcgagat ggaccattac cgcgagccgt
     2221 tcctgaatcc tgttgaccgc gagccactgt ggcgcttccc aaacgagctg ccaatcgccg
     2281 gtgagccagc gaacatcgtc gcgctggtcg aagaatacat ggactggctg caccagtccc
     2341 ctgtcccgaa gctgctgttc tggggcaccc caggcgttct gatcccaccg gccgaagccg
     2401 ctcgcctggc caaaagcctg cctaactgca aggctgtgga catcggcccg ggtctgaatc
     2461 tgctgcaaga agacaacccg gacctgatcg gcagcgagat cgcgcgctgg ctgtctactc
     2521 tggagatttc cggtcatcat caccatcacc actaatagtc aaccgctgag caataactag
     2581 cataacccct tggggcctct aaacgggtct tgaggggttt tttgctgaaa ggaggaacta
     2641 tatccggttc gcttgctgtc cataaaaccg cccagtctag ctatcgccat gtaagcccac
     2701 tgcaagctac ctgctttctc tttgcgcttg cgttttccct tgtccagata gcccagtagc
     2761 tgacattcat ccggggtcag caccgtttct gcggactggc tttctacgta atggtttctt
     2821 agacgtcagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct
     2881 aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat
     2941 attgaaaaag gaagagtatg agtattcaac atttccgtgt cgcccttatt cccttttttg
     3001 cggcattttg ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta aaagatgctg
     3061 aagatcagtt gggtgcacga gtgggttaca tcgaactgga tctcaacagc ggtaagatcc
     3121 ttgagagttt tcgccccgaa gaacgttttc caatgatgag cactttcaaa gttctgctat
     3181 gtggcgcggt attatcccgt attgacgccg ggcaagagca actcggtcgc cgcatacact
     3241 attctcagaa tgacttggtt gagtactcac cagtcacaga aaagcatctt acggatggca
     3301 tgacagtaag agaattatgc agtgctgcca taaccatgag tgataacact gcggccaact
     3361 tacttctgac aactatcgga ggaccgaagg agctaaccgc ttttttgcac aacatggggg
     3421 atcatgtaac tcgccttgat cgttgggaac cggagctgaa tgaagccata ccaaacgacg
     3481 agcgtgacac cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg
     3541 aactacttac tctagcttcc cggcaacaat taatagactg gatggaggcg gataaagttg
     3601 caggaccact tctgcgctcg gcccttccgg ctggctggtt tattgctgat aaatctggag
     3661 ccggtgagcg tgggtctcgc ggtatcattg cagcactggg gccagatggt aagccctccc
     3721 gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga
     3781 tcgctgagat aggtgcctca ctgattaagc attggtaatt cgaaatgacc gaccaagcga
     3841 cgcccaaccg gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga
     3901 taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc
     3961 cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg
     4021 ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg
     4081 aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt
     4141 tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt
     4201 gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg
     4261 cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact
     4321 ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt
     4381 cttgaagtgg tggcctaact acggctacac tagaaggaca gtatttggta tctgcgctct
     4441 gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac
     4501 cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatt
     4561 tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg
     4621 ttaagggcat atgatggtag tgtggggtct ccccatgcga gagtagggaa ctgccaggca
     4681 tcaaataaaa cgaaaggctc agtcgaaaga ctgggccttt cgttttatct gttgtttgtc
     4741 ggtgaacgct ctcctgagta ggacaaatcc gccgggagcg gatttgaacg ttgcgaagca
     4801 acggcccgga gggtggcggg caggacgccc gccataaact gccaggcatc aaattaagca
     4861 gaaggccatc ctgacggatg gcctttttgc gtttctacaa actcttttgt ttatttttct
     4921 aaatacattc aaatatgtat ccgctcatgg atctcgatcc cgcgaaatta atacgactca
     4981 ctatagggta ttttaacttt actaaggagc gatcgcttcc gaattcctac cgcggatata
     5041 tgg
//
