LOCUS       sequence-345486-        3473 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 3473)
  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 3473)
  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
FEATURES             Location/Qualifiers
     source          1..3473
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      39..66
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     primer_bind     148..167
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        148..166
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             266..286
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="EDLYFQS"
     CDS             302..1186
                     /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             1187..1204
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(1215..1233)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1229..1276
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        1505..1609
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1610..1678,1679..2470)
                     /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(1828..1847)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      complement(2628..3216)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2708..2727)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      3334..3420
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
ORIGIN
        1 aggacgcccg ccataaactg ccaggcatca aattaagcag aaggccatcc tgacggatgg
       61 cctttttgcg tttctacaaa ctcttttgtt tatttttcta aatacattca aatatgtatc
      121 cgctcatgga tctcgatccc gcgaaattaa tacgactcac tatagggtat tttaacttta
      181 ctaaggagcg atcgcttccg aattcctacc gcggatatct agatttgggc ccaattcctg
      241 caggcgagct ctcgagccaa ccactgagga tctgtacttt cagagcgata acgatggatc
      301 cgaaatcggt actggctttc cattcgaccc ccattatgtg gaagtcctgg gcgagcgcat
      361 gcactacgtc gatgttggtc cgcgcgatgg cacccctgtg ctgttcctgc acggtaaccc
      421 gacctcctcc tacgtgtggc gcaacatcat cccgcatgtt gcaccgaccc atcgctgcat
      481 tgctccagac ctgatcggta tgggcaaatc cgacaaacca gacctgggtt atttcttcga
      541 cgaccacgtc cgcttcatgg atgccttcat cgaagccctg ggtctggaag aggtcgtcct
      601 ggtcattcac gactggggct ccgctctggg tttccactgg gccaagcgca atccagagcg
      661 cgtcaaaggt attgcattta tggagttcat ccgccctatc ccgacctggg acgaatggcc
      721 agaatttgcc cgcgagacct tccaggcctt ccgcaccacc gacgtcggcc gcaagctgat
      781 catcgatcag aacgttttta tcgagggtac gctgccgatg ggtgtcgtcc gcccgctgac
      841 tgaagtcgag atggaccatt accgcgagcc gttcctgaat cctgttgacc gcgagccact
      901 gtggcgcttc ccaaacgagc tgccaatcgc cggtgagcca gcgaacatcg tcgcgctggt
      961 cgaagaatac atggactggc tgcaccagtc ccctgtcccg aagctgctgt tctggggcac
     1021 cccaggcgtt ctgatcccac cggccgaagc cgctcgcctg gccaaaagcc tgcctaactg
     1081 caaggctgtg gacatcggcc cgggtctgaa tctgctgcaa gaagacaacc cggacctgat
     1141 cggcagcgag atcgcgcgct ggctgtctac tctggagatt tccggtcatc atcaccatca
     1201 ccactaatag tcaaccgctg agcaataact agcataaccc cttggggcct ctaaacgggt
     1261 cttgaggggt tttttgctga aaggaggaac tatatccggt tcgcttgctg tccataaaac
     1321 cgcccagtct agctatcgcc atgtaagccc actgcaagct acctgctttc tctttgcgct
     1381 tgcgttttcc cttgtccaga tagcccagta gctgacattc atccggggtc agcaccgttt
     1441 ctgcggactg gctttctacg taatggtttc ttagacgtca ggtggcactt ttcggggaaa
     1501 tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat
     1561 gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagta tgagtattca
     1621 acatttccgt gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca
     1681 cccagaaacg ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac gagtgggtta
     1741 catcgaactg gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgttt
     1801 tccaatgatg agcactttca aagttctgct atgtggcgcg gtattatccc gtattgacgc
     1861 cgggcaagag caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtactc
     1921 accagtcaca gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc
     1981 cataaccatg agtgataaca ctgcggccaa cttacttctg acaactatcg gaggaccgaa
     2041 ggagctaacc gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga
     2101 accggagctg aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgtagcaat
     2161 ggcaacaacg ttgcgcaaac tattaactgg cgaactactt actctagctt cccggcaaca
     2221 attaatagac tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc
     2281 ggctggctgg tttattgctg ataaatctgg agccggtgag cgtgggtctc gcggtatcat
     2341 tgcagcactg gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag
     2401 tcaggcaact atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa
     2461 gcattggtaa ttcgaaatga ccgaccaagc gacgcccaac cggtatcagc tcactcaaag
     2521 gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa
     2581 ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc
     2641 cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca
     2701 ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg
     2761 accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct
     2821 catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt
     2881 gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag
     2941 tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc
     3001 agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac
     3061 actagaagga cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga
     3121 gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc
     3181 aagcagcaga ttacgcgcag aaaaaaagga tttcaagaag atcctttgat cttttctacg
     3241 gggtctgacg ctcagtggaa cgaaaactca cgttaagggc atatgatggt agtgtggggt
     3301 ctccccatgc gagagtaggg aactgccagg catcaaataa aacgaaaggc tcagtcgaaa
     3361 gactgggcct ttcgttttat ctgttgtttg tcggtgaacg ctctcctgag taggacaaat
     3421 ccgccgggag cggatttgaa cgttgcgaag caacggcccg gagggtggcg ggc
//
