LOCUS       sequence-345601-        4110 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 4110)
  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 4110)
  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_PRDX6-Halo
FEATURES             Location/Qualifiers
     source          1..4110
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      273..359
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      451..478
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     primer_bind     560..579
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        560..578
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             1315..1335
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="EDLYFQS"
     CDS             1351..2235
                     /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             2236..2253
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(2264..2282)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      2278..2325
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        2554..2658
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2659..2727,2728..3519)
                     /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(2877..2896)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      complement(join(3677..4110,1..155))
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3757..3776)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ctagaaggac agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag
       61 ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca
      121 agcagcagat tacgcgcaga aaaaaaggat ttcaagaaga tcctttgatc ttttctacgg
      181 ggtctgacgc tcagtggaac gaaaactcac gttaagggca tatgatggta gtgtggggtc
      241 tccccatgcg agagtaggga actgccaggc atcaaataaa acgaaaggct cagtcgaaag
      301 actgggcctt tcgttttatc tgttgtttgt cggtgaacgc tctcctgagt aggacaaatc
      361 cgccgggagc ggatttgaac gttgcgaagc aacggcccgg agggtggcgg gcaggacgcc
      421 cgccataaac tgccaggcat caaattaagc agaaggccat cctgacggat ggcctttttg
      481 cgtttctaca aactcttttg tttatttttc taaatacatt caaatatgta tccgctcatg
      541 gatctcgatc ccgcgaaatt aatacgactc actatagggt attttaactt tactaaggag
      601 cgatcgcttc cgaattccta ccgcggatat atgcccggag gtctgcttct cggggacgtg
      661 gctcccaact ttgaggccaa taccaccgtc ggccgcatcc gtttccacga ctttctggga
      721 gactcatggg gcattctctt ctcccaccct cgggacttta ccccagtgtg caccacagag
      781 cttggcagag ctgcaaagct ggcaccagaa tttgccaaga ggaatgttaa gttgattgcc
      841 ctttcaatag acagtgttga ggaccatctt gcctggagca aggatatcaa tgcttacaat
      901 tgtgaagagc ccacagaaaa gttacctttt cccatcatcg atgataggaa tcgggagctt
      961 gccatcctgt tgggcatgct ggatccagca gagaaggatg aaaagggcat gcctgtgaca
     1021 gctcgtgtgg tgtttgtttt tggtcctgat aagaagctga agctgtctat cctctaccca
     1081 gctaccactg gcaggaactt tgatgagatt ctcagggtag tcatctctct ccagctgaca
     1141 gcagaaaaaa gggttgccac cccagttgat tggaaggatg gggatagtgt gatggtcctt
     1201 ccaaccatcc ctgaagaaga agccaaaaaa cttttcccga aaggagtctt caccaaagag
     1261 ctcccatctg gcaagaaata cctccgctac acaccccagc ctgagccaac cactgaggat
     1321 ctgtactttc agagcgataa cgatggatcc gaaatcggta ctggctttcc attcgacccc
     1381 cattatgtgg aagtcctggg cgagcgcatg cactacgtcg atgttggtcc gcgcgatggc
     1441 acccctgtgc tgttcctgca cggtaacccg acctcctcct acgtgtggcg caacatcatc
     1501 ccgcatgttg caccgaccca tcgctgcatt gctccagacc tgatcggtat gggcaaatcc
     1561 gacaaaccag acctgggtta tttcttcgac gaccacgtcc gcttcatgga tgccttcatc
     1621 gaagccctgg gtctggaaga ggtcgtcctg gtcattcacg actggggctc cgctctgggt
     1681 ttccactggg ccaagcgcaa tccagagcgc gtcaaaggta ttgcatttat ggagttcatc
     1741 cgccctatcc cgacctggga cgaatggcca gaatttgccc gcgagacctt ccaggccttc
     1801 cgcaccaccg acgtcggccg caagctgatc atcgatcaga acgtttttat cgagggtacg
     1861 ctgccgatgg gtgtcgtccg cccgctgact gaagtcgaga tggaccatta ccgcgagccg
     1921 ttcctgaatc ctgttgaccg cgagccactg tggcgcttcc caaacgagct gccaatcgcc
     1981 ggtgagccag cgaacatcgt cgcgctggtc gaagaataca tggactggct gcaccagtcc
     2041 cctgtcccga agctgctgtt ctggggcacc ccaggcgttc tgatcccacc ggccgaagcc
     2101 gctcgcctgg ccaaaagcct gcctaactgc aaggctgtgg acatcggccc gggtctgaat
     2161 ctgctgcaag aagacaaccc ggacctgatc ggcagcgaga tcgcgcgctg gctgtctact
     2221 ctggagattt ccggtcatca tcaccatcac cactaatagt caaccgctga gcaataacta
     2281 gcataacccc ttggggcctc taaacgggtc ttgaggggtt ttttgctgaa aggaggaact
     2341 atatccggtt cgcttgctgt ccataaaacc gcccagtcta gctatcgcca tgtaagccca
     2401 ctgcaagcta cctgctttct ctttgcgctt gcgttttccc ttgtccagat agcccagtag
     2461 ctgacattca tccggggtca gcaccgtttc tgcggactgg ctttctacgt aatggtttct
     2521 tagacgtcag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc
     2581 taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa
     2641 tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat tccctttttt
     2701 gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct
     2761 gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag cggtaagatc
     2821 cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcactttcaa agttctgcta
     2881 tgtggcgcgg tattatcccg tattgacgcc gggcaagagc aactcggtcg ccgcatacac
     2941 tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct tacggatggc
     3001 atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac
     3061 ttacttctga caactatcgg aggaccgaag gagctaaccg cttttttgca caacatgggg
     3121 gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat accaaacgac
     3181 gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact attaactggc
     3241 gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc ggataaagtt
     3301 gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga taaatctgga
     3361 gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg taagccctcc
     3421 cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag
     3481 atcgctgaga taggtgcctc actgattaag cattggtaat tcgaaatgac cgaccaagcg
     3541 acgcccaacc ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg
     3601 ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg
     3661 ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac
     3721 gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg
     3781 gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct
     3841 ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg
     3901 tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct
     3961 gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac
     4021 tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt
     4081 tcttgaagtg gtggcctaac tacggctaca
//
