LOCUS       sequence-350741-        4056 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 4056)
  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 4056)
  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_BTF3-Halo
FEATURES             Location/Qualifiers
     source          1..4056
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             680..700
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="EDLYFQS"
     CDS             716..1600
                     /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             1601..1618
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(1629..1647)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1643..1690
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        1919..2023
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2024..2092,2093..2884)
                     /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(2242..2261)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      complement(3042..3630)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3122..3141)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      3748..3834
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      3926..3953
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     primer_bind     4035..4054
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        4035..4053
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
        1 ttttaacttt actaaggagc gatcgcttcc gaattcctac cgcggatata tgcgacggac
       61 aggcgcaccc gctcaggctg actctcgggg gcgaggtcga gccaggggcg gctgccctgg
      121 gggcgaggcg acgctgtctc aacctccacc tcgcggcgga acccgaggac aggagcctca
      181 gatgaaagaa acaatcatga accaggaaaa actcgccaaa ctgcaggcac aagtgcgcat
      241 tggtgggaaa ggaactgctc gcagaaagaa gaaggtggtt catagaacag ccacagcaga
      301 tgacaaaaaa cttcagttct ccttaaagaa gttaggggta aacaatatct ctggtattga
      361 agaggtgaat atgtttacaa accaaggaac agtgatccac tttaacaacc ctaaagttca
      421 ggcatctctg gcagcgaaca ctttcaccat tacaggccat gctgagacaa agcagctgac
      481 agaaatgcta cccagcatct taaaccagct tggtgcggat agtctgacta gtttaaggag
      541 actggccgaa gctctgccca aacaatctgt ggatggaaaa gcaccacttg ctactggaga
      601 ggatgatgat gatgaagttc cagatcttgt ggagaatttt gatgaggctt ccaagaatga
      661 ggcaaacggg ccaaccactg aggatctgta ctttcagagc gataacgatg gatccgaaat
      721 cggtactggc tttccattcg acccccatta tgtggaagtc ctgggcgagc gcatgcacta
      781 cgtcgatgtt ggtccgcgcg atggcacccc tgtgctgttc ctgcacggta acccgacctc
      841 ctcctacgtg tggcgcaaca tcatcccgca tgttgcaccg acccatcgct gcattgctcc
      901 agacctgatc ggtatgggca aatccgacaa accagacctg ggttatttct tcgacgacca
      961 cgtccgcttc atggatgcct tcatcgaagc cctgggtctg gaagaggtcg tcctggtcat
     1021 tcacgactgg ggctccgctc tgggtttcca ctgggccaag cgcaatccag agcgcgtcaa
     1081 aggtattgca tttatggagt tcatccgccc tatcccgacc tgggacgaat ggccagaatt
     1141 tgcccgcgag accttccagg ccttccgcac caccgacgtc ggccgcaagc tgatcatcga
     1201 tcagaacgtt tttatcgagg gtacgctgcc gatgggtgtc gtccgcccgc tgactgaagt
     1261 cgagatggac cattaccgcg agccgttcct gaatcctgtt gaccgcgagc cactgtggcg
     1321 cttcccaaac gagctgccaa tcgccggtga gccagcgaac atcgtcgcgc tggtcgaaga
     1381 atacatggac tggctgcacc agtcccctgt cccgaagctg ctgttctggg gcaccccagg
     1441 cgttctgatc ccaccggccg aagccgctcg cctggccaaa agcctgccta actgcaaggc
     1501 tgtggacatc ggcccgggtc tgaatctgct gcaagaagac aacccggacc tgatcggcag
     1561 cgagatcgcg cgctggctgt ctactctgga gatttccggt catcatcacc atcaccacta
     1621 atagtcaacc gctgagcaat aactagcata accccttggg gcctctaaac gggtcttgag
     1681 gggttttttg ctgaaaggag gaactatatc cggttcgctt gctgtccata aaaccgccca
     1741 gtctagctat cgccatgtaa gcccactgca agctacctgc tttctctttg cgcttgcgtt
     1801 ttcccttgtc cagatagccc agtagctgac attcatccgg ggtcagcacc gtttctgcgg
     1861 actggctttc tacgtaatgg tttcttagac gtcaggtggc acttttcggg gaaatgtgcg
     1921 cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca
     1981 ataaccctga taaatgcttc aataatattg aaaaaggaag agtatgagta ttcaacattt
     2041 ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg ctcacccaga
     2101 aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg gttacatcga
     2161 actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac gttttccaat
     2221 gatgagcact ttcaaagttc tgctatgtgg cgcggtatta tcccgtattg acgccgggca
     2281 agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt actcaccagt
     2341 cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg ctgccataac
     2401 catgagtgat aacactgcgg ccaacttact tctgacaact atcggaggac cgaaggagct
     2461 aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt gggaaccgga
     2521 gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgtag caatggcaac
     2581 aacgttgcgc aaactattaa ctggcgaact acttactcta gcttcccggc aacaattaat
     2641 agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc ttccggctgg
     2701 ctggtttatt gctgataaat ctggagccgg tgagcgtggg tctcgcggta tcattgcagc
     2761 actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg ggagtcaggc
     2821 aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga ttaagcattg
     2881 gtaattcgaa atgaccgacc aagcgacgcc caaccggtat cagctcactc aaaggcggta
     2941 atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag
     3001 caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc
     3061 cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta
     3121 taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg
     3181 ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc
     3241 tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac
     3301 gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac
     3361 ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg
     3421 aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga
     3481 aggacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt
     3541 agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag
     3601 cagattacgc gcagaaaaaa aggatttcaa gaagatcctt tgatcttttc tacggggtct
     3661 gacgctcagt ggaacgaaaa ctcacgttaa gggcatatga tggtagtgtg gggtctcccc
     3721 atgcgagagt agggaactgc caggcatcaa ataaaacgaa aggctcagtc gaaagactgg
     3781 gcctttcgtt ttatctgttg tttgtcggtg aacgctctcc tgagtaggac aaatccgccg
     3841 ggagcggatt tgaacgttgc gaagcaacgg cccggagggt ggcgggcagg acgcccgcca
     3901 taaactgcca ggcatcaaat taagcagaag gccatcctga cggatggcct ttttgcgttt
     3961 ctacaaactc ttttgtttat ttttctaaat acattcaaat atgtatccgc tcatggatct
     4021 cgatcccgcg aaattaatac gactcactat agggta
//
