LOCUS       sequence-345610-        4629 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 4629)
  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 4629)
  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_DNAJA1-Halo
FEATURES             Location/Qualifiers
     source          1..4629
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      31..58
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     primer_bind     140..159
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        140..158
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             1414..1434
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="EDLYFQS"
     CDS             1450..2334
                     /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             2335..2352
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(2363..2381)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      2377..2424
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        2653..2757
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2758..2826,2827..3618)
                     /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(2976..2995)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      complement(3776..4364)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3856..3875)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      4482..4568
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
ORIGIN
        1 cgccataaac tgccaggcat caaattaagc agaaggccat cctgacggat ggcctttttg
       61 cgtttctaca aactcttttg tttatttttc taaatacatt caaatatgta tccgctcatg
      121 gatctcgatc ccgcgaaatt aatacgactc actatagggt attttaactt tactaaggag
      181 cgatcgcttc cgaattccta ccgcggatat atggtgaaag aaacaactta ctacgatgtt
      241 ttgggggtca aacccaatgc tactcaggaa gaattgaaaa aggcttatag gaaactggct
      301 ttgaagtacc atcctgataa gaacccaaat gaaggagaga agtttaaaca gatttctcaa
      361 gcttacgaag ttctctctga tgcaaagaaa agggaattat atgacaaagg aggagaacag
      421 gcaattaaag agggtggagc aggtggcggt tttggctccc ccatggacat ctttgatatg
      481 ttttttggag gaggaggaag gatgcagaga gaaaggagag gtaaaaatgt tgtacatcag
      541 ctctcagtaa ccctagaaga cttatataat ggtgcaacaa gaaaactggc tctgcaaaag
      601 aatgtgattt gtgacaaatg tgaaggtaga ggaggtaaga aaggagcagt agagtgctgt
      661 cccaattgcc gaggtactgg aatgcaaata agaattcatc agataggacc tggaatggtt
      721 cagcaaattc agtctgtgtg catggagtgc cagggccatg gggagcggat cagtcctaaa
      781 gatagatgta aaagctgcaa cggaaggaag atagttcgag agaagaaaat tttagaagtt
      841 catattgaca aaggcatgaa agatggccag aagataacat tccatggtga aggagaccaa
      901 gaaccaggac tggagccagg cgatattatc attgtgttag atcagaagga ccatgctgtt
      961 tttactcgac gaggagaaga ccttttcatg tgtatggaca tacagctcgt tgaagcactg
     1021 tgtggcttcc agaagccaat atctactctt gacaaccgaa ccatcgtcat cacctctcat
     1081 ccaggtcaga ttgtcaagca tggagatatc aagtgtgtac taaatgaagg catgccaatt
     1141 tatcgtagac catatgaaaa gggtcgccta atcatcgaat ttaaggtaaa ctttcctgag
     1201 aatggctttc tctctcctga taaactgtct ttgctggaaa aactcctacc cgagaggaag
     1261 gaagtggaag agactgatga gatggaccaa gtagaactgg tggactttga tccaaatcag
     1321 gaaagacggc gccactacaa tggagaagca tatgaggatg atgaacatca tcccagaggt
     1381 ggtgttcagt gtcagacctc tgggccaacc actgaggatc tgtactttca gagcgataac
     1441 gatggatccg aaatcggtac tggctttcca ttcgaccccc attatgtgga agtcctgggc
     1501 gagcgcatgc actacgtcga tgttggtccg cgcgatggca cccctgtgct gttcctgcac
     1561 ggtaacccga cctcctccta cgtgtggcgc aacatcatcc cgcatgttgc accgacccat
     1621 cgctgcattg ctccagacct gatcggtatg ggcaaatccg acaaaccaga cctgggttat
     1681 ttcttcgacg accacgtccg cttcatggat gccttcatcg aagccctggg tctggaagag
     1741 gtcgtcctgg tcattcacga ctggggctcc gctctgggtt tccactgggc caagcgcaat
     1801 ccagagcgcg tcaaaggtat tgcatttatg gagttcatcc gccctatccc gacctgggac
     1861 gaatggccag aatttgcccg cgagaccttc caggccttcc gcaccaccga cgtcggccgc
     1921 aagctgatca tcgatcagaa cgtttttatc gagggtacgc tgccgatggg tgtcgtccgc
     1981 ccgctgactg aagtcgagat ggaccattac cgcgagccgt tcctgaatcc tgttgaccgc
     2041 gagccactgt ggcgcttccc aaacgagctg ccaatcgccg gtgagccagc gaacatcgtc
     2101 gcgctggtcg aagaatacat ggactggctg caccagtccc ctgtcccgaa gctgctgttc
     2161 tggggcaccc caggcgttct gatcccaccg gccgaagccg ctcgcctggc caaaagcctg
     2221 cctaactgca aggctgtgga catcggcccg ggtctgaatc tgctgcaaga agacaacccg
     2281 gacctgatcg gcagcgagat cgcgcgctgg ctgtctactc tggagatttc cggtcatcat
     2341 caccatcacc actaatagtc aaccgctgag caataactag cataacccct tggggcctct
     2401 aaacgggtct tgaggggttt tttgctgaaa ggaggaacta tatccggttc gcttgctgtc
     2461 cataaaaccg cccagtctag ctatcgccat gtaagcccac tgcaagctac ctgctttctc
     2521 tttgcgcttg cgttttccct tgtccagata gcccagtagc tgacattcat ccggggtcag
     2581 caccgtttct gcggactggc tttctacgta atggtttctt agacgtcagg tggcactttt
     2641 cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat
     2701 ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtatg
     2761 agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt
     2821 tttgctcacc cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcacga
     2881 gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa
     2941 gaacgttttc caatgatgag cactttcaaa gttctgctat gtggcgcggt attatcccgt
     3001 attgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt
     3061 gagtactcac cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc
     3121 agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac aactatcgga
     3181 ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat
     3241 cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct
     3301 gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc
     3361 cggcaacaat taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg
     3421 gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc
     3481 ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg
     3541 acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca
     3601 ctgattaagc attggtaatt cgaaatgacc gaccaagcga cgcccaaccg gtatcagctc
     3661 actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt
     3721 gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc
     3781 ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa
     3841 acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc
     3901 ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg
     3961 cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc
     4021 tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc
     4081 gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca
     4141 ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact
     4201 acggctacac tagaaggaca gtatttggta tctgcgctct gctgaagcca gttaccttcg
     4261 gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt
     4321 ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatt tcaagaagat cctttgatct
     4381 tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggcat atgatggtag
     4441 tgtggggtct ccccatgcga gagtagggaa ctgccaggca tcaaataaaa cgaaaggctc
     4501 agtcgaaaga ctgggccttt cgttttatct gttgtttgtc ggtgaacgct ctcctgagta
     4561 ggacaaatcc gccgggagcg gatttgaacg ttgcgaagca acggcccgga gggtggcggg
     4621 caggacgcc
//
