LOCUS       sequence-345558-        4518 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 4518)
  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 4518)
  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_CCDC86-Halo
FEATURES             Location/Qualifiers
     source          1..4518
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             539..559
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="EDLYFQS"
     CDS             575..1459
                     /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             1460..1477
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(1488..1506)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1502..1549
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        1778..1882
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1883..1951,1952..2743)
                     /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(2101..2120)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      complement(2901..3489)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2981..3000)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      3607..3693
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      3785..3812
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     primer_bind     3894..3913
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        3894..3912
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
        1 acctggctcc ccccggggtc agcatgagcc gagcaagcca cctccagctg gggagacggt
       61 gacaggcggc ttcggggcaa agaagcgaaa gggttcttca tcccaggccc cagcgtccaa
      121 gaagttgaat aaagaggagc ttcctgtaat cccgaagggg aagcccaaat cggggcgagt
      181 gtggaaggac cgctccaaga aaagattctc ccagatgctt caggacaagc ccctgcgcac
      241 atcgtggcag cggaagatga aggaacgaca ggagaggaag ctggccaagg actttgcccg
      301 tcacctggag gaggagaagg agaggcgccg ccaggagaag aaacagcgcc gggctgagaa
      361 cctgaaacgc cgcctggaga atgagcggaa ggcagaggtc gtccaagtga tccgaaaccc
      421 cgccaagctc aagcgggcaa agaagaagca gctgcgctcc attgagaagc gggacaccct
      481 ggccctgctg cagaagcagc cgccccagca gccggcagcc aagatcgggc caaccactga
      541 ggatctgtac tttcagagcg ataacgatgg atccgaaatc ggtactggct ttccattcga
      601 cccccattat gtggaagtcc tgggcgagcg catgcactac gtcgatgttg gtccgcgcga
      661 tggcacccct gtgctgttcc tgcacggtaa cccgacctcc tcctacgtgt ggcgcaacat
      721 catcccgcat gttgcaccga cccatcgctg cattgctcca gacctgatcg gtatgggcaa
      781 atccgacaaa ccagacctgg gttatttctt cgacgaccac gtccgcttca tggatgcctt
      841 catcgaagcc ctgggtctgg aagaggtcgt cctggtcatt cacgactggg gctccgctct
      901 gggtttccac tgggccaagc gcaatccaga gcgcgtcaaa ggtattgcat ttatggagtt
      961 catccgccct atcccgacct gggacgaatg gccagaattt gcccgcgaga ccttccaggc
     1021 cttccgcacc accgacgtcg gccgcaagct gatcatcgat cagaacgttt ttatcgaggg
     1081 tacgctgccg atgggtgtcg tccgcccgct gactgaagtc gagatggacc attaccgcga
     1141 gccgttcctg aatcctgttg accgcgagcc actgtggcgc ttcccaaacg agctgccaat
     1201 cgccggtgag ccagcgaaca tcgtcgcgct ggtcgaagaa tacatggact ggctgcacca
     1261 gtcccctgtc ccgaagctgc tgttctgggg caccccaggc gttctgatcc caccggccga
     1321 agccgctcgc ctggccaaaa gcctgcctaa ctgcaaggct gtggacatcg gcccgggtct
     1381 gaatctgctg caagaagaca acccggacct gatcggcagc gagatcgcgc gctggctgtc
     1441 tactctggag atttccggtc atcatcacca tcaccactaa tagtcaaccg ctgagcaata
     1501 actagcataa ccccttgggg cctctaaacg ggtcttgagg ggttttttgc tgaaaggagg
     1561 aactatatcc ggttcgcttg ctgtccataa aaccgcccag tctagctatc gccatgtaag
     1621 cccactgcaa gctacctgct ttctctttgc gcttgcgttt tcccttgtcc agatagccca
     1681 gtagctgaca ttcatccggg gtcagcaccg tttctgcgga ctggctttct acgtaatggt
     1741 ttcttagacg tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt
     1801 tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca
     1861 ataatattga aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc ttattccctt
     1921 ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctggtga aagtaaaaga
     1981 tgctgaagat cagttgggtg cacgagtggg ttacatcgaa ctggatctca acagcggtaa
     2041 gatccttgag agttttcgcc ccgaagaacg ttttccaatg atgagcactt tcaaagttct
     2101 gctatgtggc gcggtattat cccgtattga cgccgggcaa gagcaactcg gtcgccgcat
     2161 acactattct cagaatgact tggttgagta ctcaccagtc acagaaaagc atcttacgga
     2221 tggcatgaca gtaagagaat tatgcagtgc tgccataacc atgagtgata acactgcggc
     2281 caacttactt ctgacaacta tcggaggacc gaaggagcta accgcttttt tgcacaacat
     2341 gggggatcat gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa
     2401 cgacgagcgt gacaccacga tgcctgtagc aatggcaaca acgttgcgca aactattaac
     2461 tggcgaacta cttactctag cttcccggca acaattaata gactggatgg aggcggataa
     2521 agttgcagga ccacttctgc gctcggccct tccggctggc tggtttattg ctgataaatc
     2581 tggagccggt gagcgtgggt ctcgcggtat cattgcagca ctggggccag atggtaagcc
     2641 ctcccgtatc gtagttatct acacgacggg gagtcaggca actatggatg aacgaaatag
     2701 acagatcgct gagataggtg cctcactgat taagcattgg taattcgaaa tgaccgacca
     2761 agcgacgccc aaccggtatc agctcactca aaggcggtaa tacggttatc cacagaatca
     2821 ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa
     2881 aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat
     2941 cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc
     3001 cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc
     3061 gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt
     3121 tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac
     3181 cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg
     3241 ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca
     3301 gagttcttga agtggtggcc taactacggc tacactagaa ggacagtatt tggtatctgc
     3361 gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa
     3421 accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa
     3481 ggatttcaag aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac
     3541 tcacgttaag ggcatatgat ggtagtgtgg ggtctcccca tgcgagagta gggaactgcc
     3601 aggcatcaaa taaaacgaaa ggctcagtcg aaagactggg cctttcgttt tatctgttgt
     3661 ttgtcggtga acgctctcct gagtaggaca aatccgccgg gagcggattt gaacgttgcg
     3721 aagcaacggc ccggagggtg gcgggcagga cgcccgccat aaactgccag gcatcaaatt
     3781 aagcagaagg ccatcctgac ggatggcctt tttgcgtttc tacaaactct tttgtttatt
     3841 tttctaaata cattcaaata tgtatccgct catggatctc gatcccgcga aattaatacg
     3901 actcactata gggtatttta actttactaa ggagcgatcg cttccgaatt cctaccgcgg
     3961 atatatggat acaccgttaa ggcgcagccg acggctggga ggcctaaggc ccgaatcccc
     4021 cgagagcctc acctcagttt cgcggacgag acgggccctt gtggagttcg agtcgaaccc
     4081 agaagaaacg agggagcccg ggtctcctcc gagtgtgcag cgggctggcc tggggtcccc
     4141 cgaaaggccg ccgaagacaa gcccaggatc accccgtctg cagcagggtg caggcttgga
     4201 gtcaccccaa gggcagccag agccaggcgc agcgtccccc cagcgtcagc aagacctaca
     4261 cctggagtcg cctcaaagac agccagagta cagtcctgaa tccccacgat gtcagccgaa
     4321 gccaagtgag gaggcaccaa agtgttctca ggaccaggga gtactggcct cggagttggc
     4381 ccagaataag gaggagctga ccccgggggc cccccagcat cagctaccgc cggtcccagg
     4441 atcaccagag ccttaccccg gtcagcaagc tcccggtccg gagccctctc agccactact
     4501 ggagctgaca cccagggc
//
