LOCUS       sequence-345581-        4161 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 4161)
  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 4161)
  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_CCDC59-Halo
FEATURES             Location/Qualifiers
     source          1..4161
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      286..372
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      464..491
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     primer_bind     573..592
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        573..591
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             1379..1399
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="EDLYFQS"
     CDS             1415..2299
                     /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             2300..2317
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(2328..2346)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      2342..2389
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        2618..2722
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2723..2791,2792..3583)
                     /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(2941..2960)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      complement(join(3741..4161,1..168))
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3821..3840)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 aactacggct acactagaag gacagtattt ggtatctgcg ctctgctgaa gccagttacc
       61 ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt
      121 ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatttcaaga agatcctttg
      181 atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg gcatatgatg
      241 gtagtgtggg gtctccccat gcgagagtag ggaactgcca ggcatcaaat aaaacgaaag
      301 gctcagtcga aagactgggc ctttcgtttt atctgttgtt tgtcggtgaa cgctctcctg
      361 agtaggacaa atccgccggg agcggatttg aacgttgcga agcaacggcc cggagggtgg
      421 cgggcaggac gcccgccata aactgccagg catcaaatta agcagaaggc catcctgacg
      481 gatggccttt ttgcgtttct acaaactctt ttgtttattt ttctaaatac attcaaatat
      541 gtatccgctc atggatctcg atcccgcgaa attaatacga ctcactatag ggtattttaa
      601 ctttactaag gagcgatcgc ttccgaattc ctaccgcgga tatatggcgc cggtgaggcg
      661 gtccgcgaag tggcggcctg gtggtattga ggcgcgtggt gaaggggttt ccactgtcgg
      721 gtacaggaat aagaatgtga gacagaagac atggcggcct aaccacccgc aagccttcgt
      781 ggggagcgtt cgcgagggac aaggctttgc ttttcgaaga aaactgaaaa tacagcaaag
      841 ttacaagaaa ttgctacgga aggaaaagaa ggctcaaacg tcactggaat ctcaattcac
      901 agatcgatac ccagataatc tgaaacatct ctatttagct gaagaggaaa gacataggaa
      961 gcaagcaaga aaagtcgacc atcctttgtc agaacaagtt caccagccgt tgcttgaaga
     1021 acagtgtagc attgacgagc ctttatttga agatcagtgt agctttgacc agcctcagcc
     1081 agaagaacaa tgtattaaaa cagtaaactc ctttacaatt ccaaagaaaa ataaaaagaa
     1141 aacatcaaat caaaaagcac aagaagaata tgaacagata caagccaaac gtgctgctaa
     1201 gaaacaagaa ttcgagagga gaaaacagga gagagaagaa gcccaaaggc agtacaaaaa
     1261 gaagaaaatg gaagtgttta aaatactgaa caaaaagact aaaaagggcc aaccaaactt
     1321 gaatgtacaa atggagtacc ttcttcaaaa aatacaagaa aaatgtgggc caaccactga
     1381 ggatctgtac tttcagagcg ataacgatgg atccgaaatc ggtactggct ttccattcga
     1441 cccccattat gtggaagtcc tgggcgagcg catgcactac gtcgatgttg gtccgcgcga
     1501 tggcacccct gtgctgttcc tgcacggtaa cccgacctcc tcctacgtgt ggcgcaacat
     1561 catcccgcat gttgcaccga cccatcgctg cattgctcca gacctgatcg gtatgggcaa
     1621 atccgacaaa ccagacctgg gttatttctt cgacgaccac gtccgcttca tggatgcctt
     1681 catcgaagcc ctgggtctgg aagaggtcgt cctggtcatt cacgactggg gctccgctct
     1741 gggtttccac tgggccaagc gcaatccaga gcgcgtcaaa ggtattgcat ttatggagtt
     1801 catccgccct atcccgacct gggacgaatg gccagaattt gcccgcgaga ccttccaggc
     1861 cttccgcacc accgacgtcg gccgcaagct gatcatcgat cagaacgttt ttatcgaggg
     1921 tacgctgccg atgggtgtcg tccgcccgct gactgaagtc gagatggacc attaccgcga
     1981 gccgttcctg aatcctgttg accgcgagcc actgtggcgc ttcccaaacg agctgccaat
     2041 cgccggtgag ccagcgaaca tcgtcgcgct ggtcgaagaa tacatggact ggctgcacca
     2101 gtcccctgtc ccgaagctgc tgttctgggg caccccaggc gttctgatcc caccggccga
     2161 agccgctcgc ctggccaaaa gcctgcctaa ctgcaaggct gtggacatcg gcccgggtct
     2221 gaatctgctg caagaagaca acccggacct gatcggcagc gagatcgcgc gctggctgtc
     2281 tactctggag atttccggtc atcatcacca tcaccactaa tagtcaaccg ctgagcaata
     2341 actagcataa ccccttgggg cctctaaacg ggtcttgagg ggttttttgc tgaaaggagg
     2401 aactatatcc ggttcgcttg ctgtccataa aaccgcccag tctagctatc gccatgtaag
     2461 cccactgcaa gctacctgct ttctctttgc gcttgcgttt tcccttgtcc agatagccca
     2521 gtagctgaca ttcatccggg gtcagcaccg tttctgcgga ctggctttct acgtaatggt
     2581 ttcttagacg tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt
     2641 tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca
     2701 ataatattga aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc ttattccctt
     2761 ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctggtga aagtaaaaga
     2821 tgctgaagat cagttgggtg cacgagtggg ttacatcgaa ctggatctca acagcggtaa
     2881 gatccttgag agttttcgcc ccgaagaacg ttttccaatg atgagcactt tcaaagttct
     2941 gctatgtggc gcggtattat cccgtattga cgccgggcaa gagcaactcg gtcgccgcat
     3001 acactattct cagaatgact tggttgagta ctcaccagtc acagaaaagc atcttacgga
     3061 tggcatgaca gtaagagaat tatgcagtgc tgccataacc atgagtgata acactgcggc
     3121 caacttactt ctgacaacta tcggaggacc gaaggagcta accgcttttt tgcacaacat
     3181 gggggatcat gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa
     3241 cgacgagcgt gacaccacga tgcctgtagc aatggcaaca acgttgcgca aactattaac
     3301 tggcgaacta cttactctag cttcccggca acaattaata gactggatgg aggcggataa
     3361 agttgcagga ccacttctgc gctcggccct tccggctggc tggtttattg ctgataaatc
     3421 tggagccggt gagcgtgggt ctcgcggtat cattgcagca ctggggccag atggtaagcc
     3481 ctcccgtatc gtagttatct acacgacggg gagtcaggca actatggatg aacgaaatag
     3541 acagatcgct gagataggtg cctcactgat taagcattgg taattcgaaa tgaccgacca
     3601 agcgacgccc aaccggtatc agctcactca aaggcggtaa tacggttatc cacagaatca
     3661 ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa
     3721 aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat
     3781 cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc
     3841 cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc
     3901 gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt
     3961 tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac
     4021 cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg
     4081 ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca
     4141 gagttcttga agtggtggcc t
//
