LOCUS       sequence-344836-        4347 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 4347)
  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 4347)
  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_YBX3-Halo
FEATURES             Location/Qualifiers
     source          1..4347
                     /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             433..453
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=7xHis
                     /translation="HHHHHHH"
     CDS             1334..1354
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="EDLYFQS"
     CDS             1370..2254
                     /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             2255..2272
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(2283..2301)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      2297..2344
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        2573..2677
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2678..2746,2747..3538)
                     /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(2896..2915)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      complement(3696..4284)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3776..3795)
                     /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 atatgagtga
      421 ggcgggcgag gccaccacca ccaccaccac caccctcccg caggctccga cggaggcggc
      481 cgccgcggct ccccaggacc ccgcgcccaa gagcccggtg ggcagcggtg cgccccaggc
      541 cgcggccccg gcgcccgccg cccacgtcgc aggaaacccc ggtggggacg cggcccccgc
      601 agccacgggc accgcggccg ccgcctcttt agccaccgcc gccggcagcg aagacgcgga
      661 gaaaaaagtt ctcgccacca aagtccttgg cactgtcaaa tggttcaacg tcagaaatgg
      721 atatggattt ataaatcgaa atgacaccaa agaagatgta tttgtacatc agactgccat
      781 caagaagaat aacccacgga aatatctgcg cagtgtagga gatggagaaa ctgtagagtt
      841 tgatgtggtt gaaggagaga agggtgcaga agctgccaat gtgactggcc cggatggagt
      901 tcctgtggaa gggagtcgtt acgctgcaga tcggcgccgt tacagacgtg gctactatgg
      961 aaggcgccgt ggccctcccc ggaatgctgg tgagattgga gagatgaagg atggagtccc
     1021 agagggagca caacttcagg gaccggttca tcgaaatcca acttaccgcc caaggtaccg
     1081 tagcagggga cctcctcgcc cacgacctgc cccagcagtt ggagaggctg aagataaaga
     1141 aaatcagcaa gccaccagtg gtccaaacca gccgtctgtt cgccgtggat accggcgtcc
     1201 ctacaattac cggcgtcgcc cgcgtcctcc taacgctcct tcacaagatg gcaaagaggc
     1261 caaggcaggt gaagcaccaa ctgagaaccc tgctccaccc acccagcaga gcagtgctga
     1321 ggggccaacc actgaggatc tgtactttca gagcgataac gatggatccg aaatcggtac
     1381 tggctttcca ttcgaccccc attatgtgga agtcctgggc gagcgcatgc actacgtcga
     1441 tgttggtccg cgcgatggca cccctgtgct gttcctgcac ggtaacccga cctcctccta
     1501 cgtgtggcgc aacatcatcc cgcatgttgc accgacccat cgctgcattg ctccagacct
     1561 gatcggtatg ggcaaatccg acaaaccaga cctgggttat ttcttcgacg accacgtccg
     1621 cttcatggat gccttcatcg aagccctggg tctggaagag gtcgtcctgg tcattcacga
     1681 ctggggctcc gctctgggtt tccactgggc caagcgcaat ccagagcgcg tcaaaggtat
     1741 tgcatttatg gagttcatcc gccctatccc gacctgggac gaatggccag aatttgcccg
     1801 cgagaccttc caggccttcc gcaccaccga cgtcggccgc aagctgatca tcgatcagaa
     1861 cgtttttatc gagggtacgc tgccgatggg tgtcgtccgc ccgctgactg aagtcgagat
     1921 ggaccattac cgcgagccgt tcctgaatcc tgttgaccgc gagccactgt ggcgcttccc
     1981 aaacgagctg ccaatcgccg gtgagccagc gaacatcgtc gcgctggtcg aagaatacat
     2041 ggactggctg caccagtccc ctgtcccgaa gctgctgttc tggggcaccc caggcgttct
     2101 gatcccaccg gccgaagccg ctcgcctggc caaaagcctg cctaactgca aggctgtgga
     2161 catcggcccg ggtctgaatc tgctgcaaga agacaacccg gacctgatcg gcagcgagat
     2221 cgcgcgctgg ctgtctactc tggagatttc cggtcatcat caccatcacc actaatagtc
     2281 aaccgctgag caataactag cataacccct tggggcctct aaacgggtct tgaggggttt
     2341 tttgctgaaa ggaggaacta tatccggttc gcttgctgtc cataaaaccg cccagtctag
     2401 ctatcgccat gtaagcccac tgcaagctac ctgctttctc tttgcgcttg cgttttccct
     2461 tgtccagata gcccagtagc tgacattcat ccggggtcag caccgtttct gcggactggc
     2521 tttctacgta atggtttctt agacgtcagg tggcactttt cggggaaatg tgcgcggaac
     2581 ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc
     2641 ctgataaatg cttcaataat attgaaaaag gaagagtatg agtattcaac atttccgtgt
     2701 cgcccttatt cccttttttg cggcattttg ccttcctgtt tttgctcacc cagaaacgct
     2761 ggtgaaagta aaagatgctg aagatcagtt gggtgcacga gtgggttaca tcgaactgga
     2821 tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc caatgatgag
     2881 cactttcaaa gttctgctat gtggcgcggt attatcccgt attgacgccg ggcaagagca
     2941 actcggtcgc cgcatacact attctcagaa tgacttggtt gagtactcac cagtcacaga
     3001 aaagcatctt acggatggca tgacagtaag agaattatgc agtgctgcca taaccatgag
     3061 tgataacact gcggccaact tacttctgac aactatcgga ggaccgaagg agctaaccgc
     3121 ttttttgcac aacatggggg atcatgtaac tcgccttgat cgttgggaac cggagctgaa
     3181 tgaagccata ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg caacaacgtt
     3241 gcgcaaacta ttaactggcg aactacttac tctagcttcc cggcaacaat taatagactg
     3301 gatggaggcg gataaagttg caggaccact tctgcgctcg gcccttccgg ctggctggtt
     3361 tattgctgat aaatctggag ccggtgagcg tgggtctcgc ggtatcattg cagcactggg
     3421 gccagatggt aagccctccc gtatcgtagt tatctacacg acggggagtc aggcaactat
     3481 ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc attggtaatt
     3541 cgaaatgacc gaccaagcga cgcccaaccg gtatcagctc actcaaaggc ggtaatacgg
     3601 ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag
     3661 gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac
     3721 gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga
     3781 taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt
     3841 accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc
     3901 tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc
     3961 cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta
     4021 agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat
     4081 gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac tagaaggaca
     4141 gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct
     4201 tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt
     4261 acgcgcagaa aaaaaggatt tcaagaagat cctttgatct tttctacggg gtctgacgct
     4321 cagtggaacg aaaactcacg ttaaggg
//
