LOCUS       sequence-345582-        4107 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 4107)
  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 4107)
  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_CCDC124-Halo
FEATURES             Location/Qualifiers
     source          1..4107
                     /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             900..920
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="EDLYFQS"
     CDS             936..1820
                     /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             1821..1838
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(1849..1867)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1863..1910
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        2139..2243
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2244..2312,2313..3104)
                     /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(2462..2481)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      complement(3262..3850)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3342..3361)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      3968..4054
                     /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 gcggatatat gcccaagaag ttccagggtg
      241 agaacaccaa gtcggcagcg gcccgggcac gtagggcaga ggccaaggcg gccgctgatg
      301 ccaagaagca gaaggagctg gaggatgcct actggaagga cgacgacaaa cacgtcatga
      361 ggaaggagca gcgcaaggag gagaaggaga agcggcgcct cgaccagctg gaacgtaaga
      421 aggagacgca gcgcctactg gaggaggagg actccaagct caagggcggc aaggcgccgc
      481 gggtggccac gtccagcaag gtcacccggg cccagatcga ggacacgctg cgccgagacc
      541 atcagctcag ggaggccccg gacacagccg agaaagccaa gagccatctg gaggtgccgc
      601 tggaggagaa cgtgaaccgc cgcgtgctgg aggagggcag cgtggaggcg cgcaccatcg
      661 aggacgccat tgcagtgctc agcgtggcgg aggaggcggc cgaccggcac ccagaaagac
      721 gcatgcgggc agccttcaca gcctttgagg aagcccagct gccgcggctc aaacaagaga
      781 accccaacat gcggctgtcg cagctgaaac agctgctcaa gaaggagtgg ctccgctctc
      841 ctgacaaccc catgaaccag cgggccgtgc ccttcaatgc tcccaagggg ccaaccactg
      901 aggatctgta ctttcagagc gataacgatg gatccgaaat cggtactggc tttccattcg
      961 acccccatta tgtggaagtc ctgggcgagc gcatgcacta cgtcgatgtt ggtccgcgcg
     1021 atggcacccc tgtgctgttc ctgcacggta acccgacctc ctcctacgtg tggcgcaaca
     1081 tcatcccgca tgttgcaccg acccatcgct gcattgctcc agacctgatc ggtatgggca
     1141 aatccgacaa accagacctg ggttatttct tcgacgacca cgtccgcttc atggatgcct
     1201 tcatcgaagc cctgggtctg gaagaggtcg tcctggtcat tcacgactgg ggctccgctc
     1261 tgggtttcca ctgggccaag cgcaatccag agcgcgtcaa aggtattgca tttatggagt
     1321 tcatccgccc tatcccgacc tgggacgaat ggccagaatt tgcccgcgag accttccagg
     1381 ccttccgcac caccgacgtc ggccgcaagc tgatcatcga tcagaacgtt tttatcgagg
     1441 gtacgctgcc gatgggtgtc gtccgcccgc tgactgaagt cgagatggac cattaccgcg
     1501 agccgttcct gaatcctgtt gaccgcgagc cactgtggcg cttcccaaac gagctgccaa
     1561 tcgccggtga gccagcgaac atcgtcgcgc tggtcgaaga atacatggac tggctgcacc
     1621 agtcccctgt cccgaagctg ctgttctggg gcaccccagg cgttctgatc ccaccggccg
     1681 aagccgctcg cctggccaaa agcctgccta actgcaaggc tgtggacatc ggcccgggtc
     1741 tgaatctgct gcaagaagac aacccggacc tgatcggcag cgagatcgcg cgctggctgt
     1801 ctactctgga gatttccggt catcatcacc atcaccacta atagtcaacc gctgagcaat
     1861 aactagcata accccttggg gcctctaaac gggtcttgag gggttttttg ctgaaaggag
     1921 gaactatatc cggttcgctt gctgtccata aaaccgccca gtctagctat cgccatgtaa
     1981 gcccactgca agctacctgc tttctctttg cgcttgcgtt ttcccttgtc cagatagccc
     2041 agtagctgac attcatccgg ggtcagcacc gtttctgcgg actggctttc tacgtaatgg
     2101 tttcttagac gtcaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat
     2161 ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc
     2221 aataatattg aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc cttattccct
     2281 tttttgcggc attttgcctt cctgtttttg ctcacccaga aacgctggtg aaagtaaaag
     2341 atgctgaaga tcagttgggt gcacgagtgg gttacatcga actggatctc aacagcggta
     2401 agatccttga gagttttcgc cccgaagaac gttttccaat gatgagcact ttcaaagttc
     2461 tgctatgtgg cgcggtatta tcccgtattg acgccgggca agagcaactc ggtcgccgca
     2521 tacactattc tcagaatgac ttggttgagt actcaccagt cacagaaaag catcttacgg
     2581 atggcatgac agtaagagaa ttatgcagtg ctgccataac catgagtgat aacactgcgg
     2641 ccaacttact tctgacaact atcggaggac cgaaggagct aaccgctttt ttgcacaaca
     2701 tgggggatca tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa gccataccaa
     2761 acgacgagcg tgacaccacg atgcctgtag caatggcaac aacgttgcgc aaactattaa
     2821 ctggcgaact acttactcta gcttcccggc aacaattaat agactggatg gaggcggata
     2881 aagttgcagg accacttctg cgctcggccc ttccggctgg ctggtttatt gctgataaat
     2941 ctggagccgg tgagcgtggg tctcgcggta tcattgcagc actggggcca gatggtaagc
     3001 cctcccgtat cgtagttatc tacacgacgg ggagtcaggc aactatggat gaacgaaata
     3061 gacagatcgc tgagataggt gcctcactga ttaagcattg gtaattcgaa atgaccgacc
     3121 aagcgacgcc caaccggtat cagctcactc aaaggcggta atacggttat ccacagaatc
     3181 aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa
     3241 aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa
     3301 tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc
     3361 ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc
     3421 cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag
     3481 ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga
     3541 ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc
     3601 gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac
     3661 agagttcttg aagtggtggc ctaactacgg ctacactaga aggacagtat ttggtatctg
     3721 cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca
     3781 aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa
     3841 aggatttcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa
     3901 ctcacgttaa gggcatatga tggtagtgtg gggtctcccc atgcgagagt agggaactgc
     3961 caggcatcaa ataaaacgaa aggctcagtc gaaagactgg gcctttcgtt ttatctgttg
     4021 tttgtcggtg aacgctctcc tgagtaggac aaatccgccg ggagcggatt tgaacgttgc
     4081 gaagcaacgg cccggagggt ggcgggc
//
