LOCUS       sequence-345611-        4242 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 4242)
  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 4242)
  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_MAPRE1-Halo
FEATURES             Location/Qualifiers
     source          1..4242
                     /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             1460..1480
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="EDLYFQS"
     CDS             1496..2380
                     /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             2381..2398
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(2409..2427)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      2423..2470
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        2699..2803
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2804..2872,2873..3664)
                     /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(3022..3041)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      complement(join(3822..4242,1..168))
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3902..3921)
                     /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 tatatggcag tgaacgtata
      661 ctcaacgtca gtgaccagtg ataacctaag tcgacatgac atgctggcct ggatcaatga
      721 gtctctgcag ttgaatctga caaagatcga acagttgtgc tcaggggctg cgtattgtca
      781 gtttatggac atgctgttcc ctggctccat tgccttgaag aaagtgaaat tccaagctaa
      841 gctagaacac gagtacatcc agaacttcaa aatactacaa gcaggtttta agagaatggg
      901 tgttgacaaa ataattcctg tggacaaatt agtaaaagga aagtttcagg acaattttga
      961 attcgttcag tggttcaaga agtttttcga tgcaaactat gatggaaaag actatgaccc
     1021 tgtggctgcc agacaaggtc aagaaactgc agtggctcct tcccttgttg ctccagctct
     1081 gaataaaccg aagaaacctc tcacttctag cagtgcagct ccccagaggc ccatctcaac
     1141 acagagaacc gctgcggctc ctaaggctgg ccctggtgtg gtgcgaaaga accctggtgt
     1201 gggcaacgga gacgacgagg cagctgagtt gatgcagcag gtcaacgtat tgaaacttac
     1261 tgttgaagac ttggagaaag agagggattt ctacttcgga aagctacgga acattgaatt
     1321 gatttgccag gagaacgagg gggaaaacga ccctgtattg cagaggattg tagacattct
     1381 gtatgccaca gatgaaggct ttgtgatacc tgatgaaggg ggcccacagg aggagcaaga
     1441 agagtatgag ccaaccactg aggatctgta ctttcagagc gataacgatg gatccgaaat
     1501 cggtactggc tttccattcg acccccatta tgtggaagtc ctgggcgagc gcatgcacta
     1561 cgtcgatgtt ggtccgcgcg atggcacccc tgtgctgttc ctgcacggta acccgacctc
     1621 ctcctacgtg tggcgcaaca tcatcccgca tgttgcaccg acccatcgct gcattgctcc
     1681 agacctgatc ggtatgggca aatccgacaa accagacctg ggttatttct tcgacgacca
     1741 cgtccgcttc atggatgcct tcatcgaagc cctgggtctg gaagaggtcg tcctggtcat
     1801 tcacgactgg ggctccgctc tgggtttcca ctgggccaag cgcaatccag agcgcgtcaa
     1861 aggtattgca tttatggagt tcatccgccc tatcccgacc tgggacgaat ggccagaatt
     1921 tgcccgcgag accttccagg ccttccgcac caccgacgtc ggccgcaagc tgatcatcga
     1981 tcagaacgtt tttatcgagg gtacgctgcc gatgggtgtc gtccgcccgc tgactgaagt
     2041 cgagatggac cattaccgcg agccgttcct gaatcctgtt gaccgcgagc cactgtggcg
     2101 cttcccaaac gagctgccaa tcgccggtga gccagcgaac atcgtcgcgc tggtcgaaga
     2161 atacatggac tggctgcacc agtcccctgt cccgaagctg ctgttctggg gcaccccagg
     2221 cgttctgatc ccaccggccg aagccgctcg cctggccaaa agcctgccta actgcaaggc
     2281 tgtggacatc ggcccgggtc tgaatctgct gcaagaagac aacccggacc tgatcggcag
     2341 cgagatcgcg cgctggctgt ctactctgga gatttccggt catcatcacc atcaccacta
     2401 atagtcaacc gctgagcaat aactagcata accccttggg gcctctaaac gggtcttgag
     2461 gggttttttg ctgaaaggag gaactatatc cggttcgctt gctgtccata aaaccgccca
     2521 gtctagctat cgccatgtaa gcccactgca agctacctgc tttctctttg cgcttgcgtt
     2581 ttcccttgtc cagatagccc agtagctgac attcatccgg ggtcagcacc gtttctgcgg
     2641 actggctttc tacgtaatgg tttcttagac gtcaggtggc acttttcggg gaaatgtgcg
     2701 cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca
     2761 ataaccctga taaatgcttc aataatattg aaaaaggaag agtatgagta ttcaacattt
     2821 ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg ctcacccaga
     2881 aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg gttacatcga
     2941 actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac gttttccaat
     3001 gatgagcact ttcaaagttc tgctatgtgg cgcggtatta tcccgtattg acgccgggca
     3061 agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt actcaccagt
     3121 cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg ctgccataac
     3181 catgagtgat aacactgcgg ccaacttact tctgacaact atcggaggac cgaaggagct
     3241 aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt gggaaccgga
     3301 gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgtag caatggcaac
     3361 aacgttgcgc aaactattaa ctggcgaact acttactcta gcttcccggc aacaattaat
     3421 agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc ttccggctgg
     3481 ctggtttatt gctgataaat ctggagccgg tgagcgtggg tctcgcggta tcattgcagc
     3541 actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg ggagtcaggc
     3601 aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga ttaagcattg
     3661 gtaattcgaa atgaccgacc aagcgacgcc caaccggtat cagctcactc aaaggcggta
     3721 atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag
     3781 caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc
     3841 cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta
     3901 taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg
     3961 ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc
     4021 tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac
     4081 gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac
     4141 ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg
     4201 aggtatgtag gcggtgctac agagttcttg aagtggtggc ct
//
