LOCUS       sequence-345609-        4917 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 4917)
  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 4917)
  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_FSCN1-Halo
FEATURES             Location/Qualifiers
     source          1..4917
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      33..60
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     primer_bind     142..161
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        142..160
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             1704..1724
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="EDLYFQS"
     CDS             1740..2624
                     /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             2625..2642
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(2653..2671)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      2667..2714
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        2943..3047
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3048..3116,3117..3908)
                     /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(3266..3285)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      complement(4066..4654)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4146..4165)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      4772..4858
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
ORIGIN
        1 cccgccataa actgccaggc atcaaattaa gcagaaggcc atcctgacgg atggcctttt
       61 tgcgtttcta caaactcttt tgtttatttt tctaaataca ttcaaatatg tatccgctca
      121 tggatctcga tcccgcgaaa ttaatacgac tcactatagg gtattttaac tttactaagg
      181 agcgatcgct tccgaattcc taccgcggat atatgaccgc caacggcaca gccgaggcgg
      241 tgcagatcca gttcggcctc atcaactgcg gcaacaagta cctgacggcc gaggcgttcg
      301 ggttcaaggt gaacgcgtcc gccagcagcc tgaagaagaa gcagatctgg acgctggagc
      361 agccccctga cgaggcgggc agcgcggccg tgtgcctgcg cagccacctg ggccgctacc
      421 tggcggcgga caaggacggc aacgtgacct gcgagcgcga ggtgcccggt cccgactgcc
      481 gtttcctcat cgtggcgcac gacgacggtc gctggtcgct gcagtccgag gcgcaccggc
      541 gctacttcgg cggcaccgag gaccgcctgt cctgcttcgc gcagacggtg tcccccgccg
      601 agaagtggag cgtgcacatc gccatgcacc ctcaggtcaa catctacagc gtcacccgta
      661 agcgctacgc gcacctgagc gcgcggccgg ccgacgagat cgccgtggac cgcgacgtgc
      721 cctggggcgt cgactcgctc atcaccctcg ccttccagga ccagcgctac agcgtgcaga
      781 ccgccgacca ccgcttcctg cgccacgacg ggcgcctggt ggcgcgcccc gagccggcca
      841 ctggctacac gctggagttc cgctccggca aggtggcctt ccgcgactgc gagggccgtt
      901 acctggcgcc gtcggggccc agcggcacgc tcaaggcggg caaggccacc aaggtgggca
      961 aggacgagct ctttgctctg gagcagagct gcgcccaggt cgtgctgcag gcggccaacg
     1021 agaggaacgt gtccacgcgc cagggtatgg acctgtctgc caatcaggac gaggagaccg
     1081 accaggagac cttccagctg gagatcgacc gcgacaccaa aaagtgtgcc ttccgtaccc
     1141 acacgggcaa gtactggacg ctgacggcca ccgggggcgt gcagtccacc gcctccagca
     1201 agaatgccag ctgctacttc gacatcgagt ggcgtgaccg gcgcatcaca ctgagggcgt
     1261 ccaatggcaa gtttgtgacc tccaagaaga atgggcagct ggccgcctcg gtggagacag
     1321 caggggactc agagctcttc ctcatgaagc tcatcaaccg ccccatcatc gtgttccgcg
     1381 gggagcatgg cttcatcggc tgccgcaagg tcacgggcac cctggacgcc aaccgctcca
     1441 gctatgacgt cttccagctg gagttcaacg atggcgccta caacatcaaa gactccacag
     1501 gcaaatactg gacggtgggc agtgactccg cggtcaccag cagcggcgac actcctgtgg
     1561 acttcttctt cgagttctgc gactataaca aggtggccat caaggtgggc gggcgctacc
     1621 tgaagggcga ccacgcaggc gtcctgaagg cctcggcgga aaccgtggac cccgcctcgc
     1681 tctgggagta cgggccaacc actgaggatc tgtactttca gagcgataac gatggatccg
     1741 aaatcggtac tggctttcca ttcgaccccc attatgtgga agtcctgggc gagcgcatgc
     1801 actacgtcga tgttggtccg cgcgatggca cccctgtgct gttcctgcac ggtaacccga
     1861 cctcctccta cgtgtggcgc aacatcatcc cgcatgttgc accgacccat cgctgcattg
     1921 ctccagacct gatcggtatg ggcaaatccg acaaaccaga cctgggttat ttcttcgacg
     1981 accacgtccg cttcatggat gccttcatcg aagccctggg tctggaagag gtcgtcctgg
     2041 tcattcacga ctggggctcc gctctgggtt tccactgggc caagcgcaat ccagagcgcg
     2101 tcaaaggtat tgcatttatg gagttcatcc gccctatccc gacctgggac gaatggccag
     2161 aatttgcccg cgagaccttc caggccttcc gcaccaccga cgtcggccgc aagctgatca
     2221 tcgatcagaa cgtttttatc gagggtacgc tgccgatggg tgtcgtccgc ccgctgactg
     2281 aagtcgagat ggaccattac cgcgagccgt tcctgaatcc tgttgaccgc gagccactgt
     2341 ggcgcttccc aaacgagctg ccaatcgccg gtgagccagc gaacatcgtc gcgctggtcg
     2401 aagaatacat ggactggctg caccagtccc ctgtcccgaa gctgctgttc tggggcaccc
     2461 caggcgttct gatcccaccg gccgaagccg ctcgcctggc caaaagcctg cctaactgca
     2521 aggctgtgga catcggcccg ggtctgaatc tgctgcaaga agacaacccg gacctgatcg
     2581 gcagcgagat cgcgcgctgg ctgtctactc tggagatttc cggtcatcat caccatcacc
     2641 actaatagtc aaccgctgag caataactag cataacccct tggggcctct aaacgggtct
     2701 tgaggggttt tttgctgaaa ggaggaacta tatccggttc gcttgctgtc cataaaaccg
     2761 cccagtctag ctatcgccat gtaagcccac tgcaagctac ctgctttctc tttgcgcttg
     2821 cgttttccct tgtccagata gcccagtagc tgacattcat ccggggtcag caccgtttct
     2881 gcggactggc tttctacgta atggtttctt agacgtcagg tggcactttt cggggaaatg
     2941 tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga
     3001 gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtatg agtattcaac
     3061 atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt tttgctcacc
     3121 cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcacga gtgggttaca
     3181 tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc
     3241 caatgatgag cactttcaaa gttctgctat gtggcgcggt attatcccgt attgacgccg
     3301 ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt gagtactcac
     3361 cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc agtgctgcca
     3421 taaccatgag tgataacact gcggccaact tacttctgac aactatcgga ggaccgaagg
     3481 agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat cgttgggaac
     3541 cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg
     3601 caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc cggcaacaat
     3661 taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg gcccttccgg
     3721 ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc ggtatcattg
     3781 cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg acggggagtc
     3841 aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc
     3901 attggtaatt cgaaatgacc gaccaagcga cgcccaaccg gtatcagctc actcaaaggc
     3961 ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg
     4021 ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg
     4081 cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg
     4141 actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac
     4201 cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca
     4261 tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt
     4321 gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc
     4381 caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag
     4441 agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac
     4501 tagaaggaca gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt
     4561 tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa
     4621 gcagcagatt acgcgcagaa aaaaaggatt tcaagaagat cctttgatct tttctacggg
     4681 gtctgacgct cagtggaacg aaaactcacg ttaagggcat atgatggtag tgtggggtct
     4741 ccccatgcga gagtagggaa ctgccaggca tcaaataaaa cgaaaggctc agtcgaaaga
     4801 ctgggccttt cgttttatct gttgtttgtc ggtgaacgct ctcctgagta ggacaaatcc
     4861 gccgggagcg gatttgaacg ttgcgaagca acggcccgga gggtggcggg caggacg
//
