LOCUS       sequence-344835-        5841 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 5841)
  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 5841)
  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_BRD2-Halo
FEATURES             Location/Qualifiers
     source          1..5841
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      55..141
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      233..260
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     primer_bind     342..361
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        342..360
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             2828..2848
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="EDLYFQS"
     CDS             2864..3748
                     /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             3749..3766
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(3777..3795)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      3791..3838
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        4067..4171
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4172..4240,4241..5032)
                     /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(4390..4409)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      complement(5190..5778)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5270..5289)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 catatgatgg tagtgtgggg tctccccatg cgagagtagg gaactgccag gcatcaaata
       61 aaacgaaagg ctcagtcgaa agactgggcc tttcgtttta tctgttgttt gtcggtgaac
      121 gctctcctga gtaggacaaa tccgccggga gcggatttga acgttgcgaa gcaacggccc
      181 ggagggtggc gggcaggacg cccgccataa actgccaggc atcaaattaa gcagaaggcc
      241 atcctgacgg atggcctttt tgcgtttcta caaactcttt tgtttatttt tctaaataca
      301 ttcaaatatg tatccgctca tggatctcga tcccgcgaaa ttaatacgac tcactatagg
      361 gtattttaac tttactaagg agcgatcgct tccgaattcc taccgcggat atatgctgca
      421 aaacgtgact ccccacaata agctccctgg ggaagggaat gcagggttgc tggggctggg
      481 cccagaagca gcagcaccag ggaaaaggat tcgaaaaccc tctctcttgt atgagggctt
      541 tgagagcccc acaatggctt cggtgcctgc tttgcaactt acccctgcca acccaccacc
      601 cccggaggtg tccaatccca aaaagccagg acgagttacc aaccagctgc aatacctaca
      661 caaggtagtg atgaaggctc tgtggaaaca tcagttcgca tggccattcc ggcagcctgt
      721 ggatgctgtc aaactgggtc taccggatta tcacaaaatt ataaaacagc ctatggacat
      781 gggtactatt aagaggagac ttgaaaacaa ttattattgg gctgcttcag agtgtatgca
      841 agattttaat accatgttca ccaactgtta catttacaac aagcccactg atgatattgt
      901 cctaatggca caaacgctgg aaaagatatt cctacagaag gttgcatcaa tgccacaaga
      961 agaacaagag ctggtagtga ccatccctaa gaacagccac aagaaggggg ccaagttggc
     1021 agcgctccag ggcagtgtta ccagtgccca tcaggtgcct gccgtctctt ctgtgtcaca
     1081 cacagccctg tatactcctc cacctgagat acctaccact gtcctcaaca ttccccaccc
     1141 atcagtcatt tcctctccac ttctcaagtc cttgcactct gctggacccc cgctccttgc
     1201 tgttactgca gctcctccag cccagcccct tgccaagaaa aaaggcgtaa agcggaaagc
     1261 agatactacc acccctacac ctacagccat cttggctcct ggttctccag ctagccctcc
     1321 tgggagtctt gagcctaagg cagcacggct tccccctatg cgtagagaga gtggtcgccc
     1381 catcaagccc ccacgcaaag acttgcctga ctctcagcaa caacaccaga gctctaagaa
     1441 aggaaagctt tcagaacagt taaaacattg caatggcatt ttgaaggagt tactctctaa
     1501 gaagcatgct gcctatgctt ggcctttcta taaaccagtg gatgcttctg cacttggcct
     1561 gcatgactac catgacatca ttaagcaccc catggacctc agcactgtca agcggaagat
     1621 ggagaaccgt gattaccggg atgcacagga gtttgctgct gatgtacggc ttatgttctc
     1681 caactgctat aagtacaatc ccccagatca cgatgttgtg gcaatggcac gaaagctaca
     1741 ggatgtattt gagttccgtt atgccaagat gccagatgaa ccactagaac cagggccttt
     1801 accagtctct actgccatgc cccctggctt ggccaaatcg tcttcagagt cctccagtga
     1861 ggaaagtagc agtgagagct cctctgagga agaggaggag gaagatgagg aggacgagga
     1921 ggaagaagag agtgaaagct cagactcaga ggaagaaagg gctcatcgct tagcagaact
     1981 acaggaacag cttcgggcag tacatgaaca actggctgct ctgtcccagg gtccaatatc
     2041 caagcccaag aggaaaagag agaaaaaaga gaaaaagaag aaacggaagg cagagaagca
     2101 tcgaggccga gctggggccg atgaagatga caaggggcct agggcacccc gcccacctca
     2161 acctaagaag tccaagaaag caagtggcag tgggggtggc agtgctgctt taggcccttc
     2221 tggctttgga ccttctggag gaagtggcac caagctcccc aaaaaggcca caaagacagc
     2281 cccacctgcc ctgcctacag gttatgattc agaggaggag gaagagagca ggcccatgag
     2341 ttacgatgag aagcggcagc tgagcctgga catcaacaaa ttacctgggg agaagctggg
     2401 ccgagttgtg catataatcc aagccaggga gccctcttta cgtgattcaa acccagaaga
     2461 gattgagatt gattttgaaa cactcaagcc atccacactt agagagcttg agcgctatgt
     2521 cctttcctgc ctacgtaaga aaccccggaa gccctacacc attaagaagc ctgtgggaaa
     2581 gacaaaggag gaactggctt tggagaaaaa gcgggaatta gaaaagcggt tacaagatgt
     2641 cagcggacag ctcaattcta ctaaaaagcc ccccaagaaa gcgaatgaga aaacagagtc
     2701 atcctctgca cagcaagtag cagtgtcacg ccttagcgct tccagctcca gctcagattc
     2761 cagctcctcc tcttcctcgt cgtcgtcttc agacaccagt gattcagact caggcgggcc
     2821 aaccactgag gatctgtact ttcagagcga taacgatgga tccgaaatcg gtactggctt
     2881 tccattcgac ccccattatg tggaagtcct gggcgagcgc atgcactacg tcgatgttgg
     2941 tccgcgcgat ggcacccctg tgctgttcct gcacggtaac ccgacctcct cctacgtgtg
     3001 gcgcaacatc atcccgcatg ttgcaccgac ccatcgctgc attgctccag acctgatcgg
     3061 tatgggcaaa tccgacaaac cagacctggg ttatttcttc gacgaccacg tccgcttcat
     3121 ggatgccttc atcgaagccc tgggtctgga agaggtcgtc ctggtcattc acgactgggg
     3181 ctccgctctg ggtttccact gggccaagcg caatccagag cgcgtcaaag gtattgcatt
     3241 tatggagttc atccgcccta tcccgacctg ggacgaatgg ccagaatttg cccgcgagac
     3301 cttccaggcc ttccgcacca ccgacgtcgg ccgcaagctg atcatcgatc agaacgtttt
     3361 tatcgagggt acgctgccga tgggtgtcgt ccgcccgctg actgaagtcg agatggacca
     3421 ttaccgcgag ccgttcctga atcctgttga ccgcgagcca ctgtggcgct tcccaaacga
     3481 gctgccaatc gccggtgagc cagcgaacat cgtcgcgctg gtcgaagaat acatggactg
     3541 gctgcaccag tcccctgtcc cgaagctgct gttctggggc accccaggcg ttctgatccc
     3601 accggccgaa gccgctcgcc tggccaaaag cctgcctaac tgcaaggctg tggacatcgg
     3661 cccgggtctg aatctgctgc aagaagacaa cccggacctg atcggcagcg agatcgcgcg
     3721 ctggctgtct actctggaga tttccggtca tcatcaccat caccactaat agtcaaccgc
     3781 tgagcaataa ctagcataac cccttggggc ctctaaacgg gtcttgaggg gttttttgct
     3841 gaaaggagga actatatccg gttcgcttgc tgtccataaa accgcccagt ctagctatcg
     3901 ccatgtaagc ccactgcaag ctacctgctt tctctttgcg cttgcgtttt cccttgtcca
     3961 gatagcccag tagctgacat tcatccgggg tcagcaccgt ttctgcggac tggctttcta
     4021 cgtaatggtt tcttagacgt caggtggcac ttttcgggga aatgtgcgcg gaacccctat
     4081 ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata
     4141 aatgcttcaa taatattgaa aaaggaagag tatgagtatt caacatttcc gtgtcgccct
     4201 tattcccttt tttgcggcat tttgccttcc tgtttttgct cacccagaaa cgctggtgaa
     4261 agtaaaagat gctgaagatc agttgggtgc acgagtgggt tacatcgaac tggatctcaa
     4321 cagcggtaag atccttgaga gttttcgccc cgaagaacgt tttccaatga tgagcacttt
     4381 caaagttctg ctatgtggcg cggtattatc ccgtattgac gccgggcaag agcaactcgg
     4441 tcgccgcata cactattctc agaatgactt ggttgagtac tcaccagtca cagaaaagca
     4501 tcttacggat ggcatgacag taagagaatt atgcagtgct gccataacca tgagtgataa
     4561 cactgcggcc aacttacttc tgacaactat cggaggaccg aaggagctaa ccgctttttt
     4621 gcacaacatg ggggatcatg taactcgcct tgatcgttgg gaaccggagc tgaatgaagc
     4681 cataccaaac gacgagcgtg acaccacgat gcctgtagca atggcaacaa cgttgcgcaa
     4741 actattaact ggcgaactac ttactctagc ttcccggcaa caattaatag actggatgga
     4801 ggcggataaa gttgcaggac cacttctgcg ctcggccctt ccggctggct ggtttattgc
     4861 tgataaatct ggagccggtg agcgtgggtc tcgcggtatc attgcagcac tggggccaga
     4921 tggtaagccc tcccgtatcg tagttatcta cacgacgggg agtcaggcaa ctatggatga
     4981 acgaaataga cagatcgctg agataggtgc ctcactgatt aagcattggt aattcgaaat
     5041 gaccgaccaa gcgacgccca accggtatca gctcactcaa aggcggtaat acggttatcc
     5101 acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg
     5161 aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat
     5221 cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag
     5281 gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga
     5341 tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg
     5401 tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt
     5461 cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac
     5521 gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc
     5581 ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag gacagtattt
     5641 ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc
     5701 ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc
     5761 agaaaaaaag gatttcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg
     5821 aacgaaaact cacgttaagg g
//
