LOCUS       sequence-345074-        5307 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 5307)
  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 5307)
  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_TKT-Halo
FEATURES             Location/Qualifiers
     source          1..5307
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      65..151
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      243..270
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     primer_bind     352..371
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        352..370
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             2304..2324
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="EDLYFQS"
     CDS             2340..3224
                     /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             3225..3242
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(3253..3271)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      3267..3314
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        3543..3647
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3648..3716,3717..4508)
                     /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(3866..3885)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      complement(4666..5254)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4746..4765)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 acgttaaggg catatgatgg tagtgtgggg tctccccatg cgagagtagg gaactgccag
       61 gcatcaaata aaacgaaagg ctcagtcgaa agactgggcc tttcgtttta tctgttgttt
      121 gtcggtgaac gctctcctga gtaggacaaa tccgccggga gcggatttga acgttgcgaa
      181 gcaacggccc ggagggtggc gggcaggacg cccgccataa actgccaggc atcaaattaa
      241 gcagaaggcc atcctgacgg atggcctttt tgcgtttcta caaactcttt tgtttatttt
      301 tctaaataca ttcaaatatg tatccgctca tggatctcga tcccgcgaaa ttaatacgac
      361 tcactatagg gtattttaac tttactaagg agcgatcgct tccgaattcc taccgcggat
      421 atatggagag ctaccacaag cctgaccagc agaagctgca ggccttgaag gacacggcca
      481 accgcctacg tatcagctcc atccaggcca ccactgcggc gggctctggc caccccacgt
      541 catgctgcag cgccgcagag atcatggctg tcctcttttt ccacaccatg cgctacaagt
      601 cccaggaccc ccggaatccg cacaatgacc gctttgtgct ctccaagggc catgcagctc
      661 ccatcctcta cgcggtctgg gctgaagctg gtttcctggc cgaggcggag ctgctgaacc
      721 tgaggaagat cagctccgac ttggacgggc acccggtccc gaaacaagct ttcaccgacg
      781 tggccactgg ctccctgggc cagggcctcg gggccgcttg tgggatggcc tacaccggca
      841 aatacttcga caaggccagc taccgagtct attgcttgct gggagacggg gagctgtcag
      901 agggctctgt atgggaggcc atggccttcg ccagcatcta taagctggac aaccttgtgg
      961 ccattctaga catcaatcgc ctgggccaga gtgacccggc cccactgcag caccagatgg
     1021 acatctacca gaagcggtgc gaggccttcg gttggcatgc catcatcgtg gatggacaca
     1081 gcgtggagga gctgtgcaag gcctttggcc aggccaagca ccagccaaca gccatcattg
     1141 ccaagacctt caagggccga gggatcacgg gggtagaaga taaggagtct tggcatggga
     1201 agcccctccc caaaaacatg gctgagcaga tcatccagga gatctacagc cagatccaga
     1261 gcaaaaagaa gatcctggca acccctccac aggaggacgc accctcagtg gacattgcca
     1321 acatccgcat gcccagcctg cccagctaca aagttgggga caagatagcc acccgcaagg
     1381 cctacgggca ggcactggcc aagctgggcc atgccagtga ccgcatcatc gccctggatg
     1441 gggacaccaa aaattccacc ttctcggaga tcttcaaaaa ggagcacccg gaccgcttca
     1501 tcgagtgcta cattgctgag cagaacatgg tgagcatcgc ggtgggctgt gccacccgca
     1561 acaggacggt gcccttctgc agcacttttg cagccttctt cacgcgggcc tttgaccaga
     1621 ttcgcatggc cgccatctcc gagagcaaca tcaacctctg cggctcccac tgcggcgttt
     1681 ccatcgggga agacgggccc tcccagatgg ccctagaaga tctggctatg tttcggtcag
     1741 tccccacatc aactgtcttt tacccaagtg atggcgttgc tacagagaag gcagtggaac
     1801 tagccgccaa tacaaagggt atctgcttca tccggaccag ccgcccagaa aatgccatca
     1861 tctataacaa caatgaggac ttccaggtcg gacaagccaa ggtggtcctg aagagcaagg
     1921 atgaccaggt gaccgttatc ggggctgggg tgaccctgca cgaggccttg gccgctgccg
     1981 aactgctgaa gaaagaaaag atcaacatcc gcgtgctgga ccccttcacc atcaagcccc
     2041 tggacagaaa actcattctc gacagcgctc gtgccaccaa gggcaggatc ctcaccgtgg
     2101 aggaccatta ttatgaaggt ggcattggtg aggctgtgtc cagtgcagta gtgggcgagc
     2161 ctggcatcac tgtcacccac ctggcagtta accgggtacc aagaagtggg aagccggctg
     2221 agctgctgaa gatgtttggt atcgacaggg atgccattgc acaagctgtg aggggcctca
     2281 tcaccaaggc cgggccaacc actgaggatc tgtactttca gagcgataac gatggatccg
     2341 aaatcggtac tggctttcca ttcgaccccc attatgtgga agtcctgggc gagcgcatgc
     2401 actacgtcga tgttggtccg cgcgatggca cccctgtgct gttcctgcac ggtaacccga
     2461 cctcctccta cgtgtggcgc aacatcatcc cgcatgttgc accgacccat cgctgcattg
     2521 ctccagacct gatcggtatg ggcaaatccg acaaaccaga cctgggttat ttcttcgacg
     2581 accacgtccg cttcatggat gccttcatcg aagccctggg tctggaagag gtcgtcctgg
     2641 tcattcacga ctggggctcc gctctgggtt tccactgggc caagcgcaat ccagagcgcg
     2701 tcaaaggtat tgcatttatg gagttcatcc gccctatccc gacctgggac gaatggccag
     2761 aatttgcccg cgagaccttc caggccttcc gcaccaccga cgtcggccgc aagctgatca
     2821 tcgatcagaa cgtttttatc gagggtacgc tgccgatggg tgtcgtccgc ccgctgactg
     2881 aagtcgagat ggaccattac cgcgagccgt tcctgaatcc tgttgaccgc gagccactgt
     2941 ggcgcttccc aaacgagctg ccaatcgccg gtgagccagc gaacatcgtc gcgctggtcg
     3001 aagaatacat ggactggctg caccagtccc ctgtcccgaa gctgctgttc tggggcaccc
     3061 caggcgttct gatcccaccg gccgaagccg ctcgcctggc caaaagcctg cctaactgca
     3121 aggctgtgga catcggcccg ggtctgaatc tgctgcaaga agacaacccg gacctgatcg
     3181 gcagcgagat cgcgcgctgg ctgtctactc tggagatttc cggtcatcat caccatcacc
     3241 actaatagtc aaccgctgag caataactag cataacccct tggggcctct aaacgggtct
     3301 tgaggggttt tttgctgaaa ggaggaacta tatccggttc gcttgctgtc cataaaaccg
     3361 cccagtctag ctatcgccat gtaagcccac tgcaagctac ctgctttctc tttgcgcttg
     3421 cgttttccct tgtccagata gcccagtagc tgacattcat ccggggtcag caccgtttct
     3481 gcggactggc tttctacgta atggtttctt agacgtcagg tggcactttt cggggaaatg
     3541 tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga
     3601 gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtatg agtattcaac
     3661 atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt tttgctcacc
     3721 cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcacga gtgggttaca
     3781 tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc
     3841 caatgatgag cactttcaaa gttctgctat gtggcgcggt attatcccgt attgacgccg
     3901 ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt gagtactcac
     3961 cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc agtgctgcca
     4021 taaccatgag tgataacact gcggccaact tacttctgac aactatcgga ggaccgaagg
     4081 agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat cgttgggaac
     4141 cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg
     4201 caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc cggcaacaat
     4261 taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg gcccttccgg
     4321 ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc ggtatcattg
     4381 cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg acggggagtc
     4441 aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc
     4501 attggtaatt cgaaatgacc gaccaagcga cgcccaaccg gtatcagctc actcaaaggc
     4561 ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg
     4621 ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg
     4681 cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg
     4741 actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac
     4801 cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca
     4861 tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt
     4921 gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc
     4981 caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag
     5041 agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac
     5101 tagaaggaca gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt
     5161 tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa
     5221 gcagcagatt acgcgcagaa aaaaaggatt tcaagaagat cctttgatct tttctacggg
     5281 gtctgacgct cagtggaacg aaaactc
//
