LOCUS       sequence-344840-        3441 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 3441)
  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 3441)
  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_minus_cer-region_minus_EcoRV-XhoI
FEATURES             Location/Qualifiers
     source          1..3441
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      244..330
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      422..449
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     primer_bind     531..550
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        531..549
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             617..637
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="EDLYFQS"
     CDS             653..1537
                     /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             1538..1555
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(1566..1584)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1580..1627
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        1856..1960
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1961..2029,2030..2821)
                     /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(2179..2198)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      complement(join(2979..3441,1..126))
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3059..3078)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc
       61 accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga
      121 tttcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca
      181 cgttaagggc atatgatggt agtgtggggt ctccccatgc gagagtaggg aactgccagg
      241 catcaaataa aacgaaaggc tcagtcgaaa gactgggcct ttcgttttat ctgttgtttg
      301 tcggtgaacg ctctcctgag taggacaaat ccgccgggag cggatttgaa cgttgcgaag
      361 caacggcccg gagggtggcg ggcaggacgc ccgccataaa ctgccaggca tcaaattaag
      421 cagaaggcca tcctgacgga tggccttttt gcgtttctac aaactctttt gtttattttt
      481 ctaaatacat tcaaatatgt atccgctcat ggatctcgat cccgcgaaat taatacgact
      541 cactataggg tattttaact ttactaagga gcgatcgctt ccgaattcct accgcggata
      601 tatggggcca accactgagg atctgtactt tcagagcgat aacgatggat ccgaaatcgg
      661 tactggcttt ccattcgacc cccattatgt ggaagtcctg ggcgagcgca tgcactacgt
      721 cgatgttggt ccgcgcgatg gcacccctgt gctgttcctg cacggtaacc cgacctcctc
      781 ctacgtgtgg cgcaacatca tcccgcatgt tgcaccgacc catcgctgca ttgctccaga
      841 cctgatcggt atgggcaaat ccgacaaacc agacctgggt tatttcttcg acgaccacgt
      901 ccgcttcatg gatgccttca tcgaagccct gggtctggaa gaggtcgtcc tggtcattca
      961 cgactggggc tccgctctgg gtttccactg ggccaagcgc aatccagagc gcgtcaaagg
     1021 tattgcattt atggagttca tccgccctat cccgacctgg gacgaatggc cagaatttgc
     1081 ccgcgagacc ttccaggcct tccgcaccac cgacgtcggc cgcaagctga tcatcgatca
     1141 gaacgttttt atcgagggta cgctgccgat gggtgtcgtc cgcccgctga ctgaagtcga
     1201 gatggaccat taccgcgagc cgttcctgaa tcctgttgac cgcgagccac tgtggcgctt
     1261 cccaaacgag ctgccaatcg ccggtgagcc agcgaacatc gtcgcgctgg tcgaagaata
     1321 catggactgg ctgcaccagt cccctgtccc gaagctgctg ttctggggca ccccaggcgt
     1381 tctgatccca ccggccgaag ccgctcgcct ggccaaaagc ctgcctaact gcaaggctgt
     1441 ggacatcggc ccgggtctga atctgctgca agaagacaac ccggacctga tcggcagcga
     1501 gatcgcgcgc tggctgtcta ctctggagat ttccggtcat catcaccatc accactaata
     1561 gtcaaccgct gagcaataac tagcataacc ccttggggcc tctaaacggg tcttgagggg
     1621 ttttttgctg aaaggaggaa ctatatccgg ttcgcttgct gtccataaaa ccgcccagtc
     1681 tagctatcgc catgtaagcc cactgcaagc tacctgcttt ctctttgcgc ttgcgttttc
     1741 ccttgtccag atagcccagt agctgacatt catccggggt cagcaccgtt tctgcggact
     1801 ggctttctac gtaatggttt cttagacgtc aggtggcact tttcggggaa atgtgcgcgg
     1861 aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata
     1921 accctgataa atgcttcaat aatattgaaa aaggaagagt atgagtattc aacatttccg
     1981 tgtcgccctt attccctttt ttgcggcatt ttgccttcct gtttttgctc acccagaaac
     2041 gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt acatcgaact
     2101 ggatctcaac agcggtaaga tccttgagag ttttcgcccc gaagaacgtt ttccaatgat
     2161 gagcactttc aaagttctgc tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga
     2221 gcaactcggt cgccgcatac actattctca gaatgacttg gttgagtact caccagtcac
     2281 agaaaagcat cttacggatg gcatgacagt aagagaatta tgcagtgctg ccataaccat
     2341 gagtgataac actgcggcca acttacttct gacaactatc ggaggaccga aggagctaac
     2401 cgcttttttg cacaacatgg gggatcatgt aactcgcctt gatcgttggg aaccggagct
     2461 gaatgaagcc ataccaaacg acgagcgtga caccacgatg cctgtagcaa tggcaacaac
     2521 gttgcgcaaa ctattaactg gcgaactact tactctagct tcccggcaac aattaataga
     2581 ctggatggag gcggataaag ttgcaggacc acttctgcgc tcggcccttc cggctggctg
     2641 gtttattgct gataaatctg gagccggtga gcgtgggtct cgcggtatca ttgcagcact
     2701 ggggccagat ggtaagccct cccgtatcgt agttatctac acgacgggga gtcaggcaac
     2761 tatggatgaa cgaaatagac agatcgctga gataggtgcc tcactgatta agcattggta
     2821 attcgaaatg accgaccaag cgacgcccaa ccggtatcag ctcactcaaa ggcggtaata
     2881 cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa
     2941 aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct
     3001 gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa
     3061 agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg
     3121 cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca
     3181 cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa
     3241 ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg
     3301 gtaagacacg acttatcgcc actggcagca gccactggta acaggattag cagagcgagg
     3361 tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta cactagaagg
     3421 acagtatttg gtatctgcgc t
//
