LOCUS       sequence-345605-        4611 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 4611)
  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 4611)
  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_NAP1L1-Halo
FEATURES             Location/Qualifiers
     source          1..4611
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      307..393
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      485..512
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     primer_bind     594..613
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        594..612
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             1850..1870
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="EDLYFQS"
     CDS             1886..2770
                     /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             2771..2788
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(2799..2817)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      2813..2860
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        3089..3193
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3194..3262,3263..4054)
                     /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(3412..3431)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      complement(join(4212..4611,1..189))
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4292..4311)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gagttcttga agtggtggcc taactacggc tacactagaa ggacagtatt tggtatctgc
       61 gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa
      121 accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa
      181 ggatttcaag aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac
      241 tcacgttaag ggcatatgat ggtagtgtgg ggtctcccca tgcgagagta gggaactgcc
      301 aggcatcaaa taaaacgaaa ggctcagtcg aaagactggg cctttcgttt tatctgttgt
      361 ttgtcggtga acgctctcct gagtaggaca aatccgccgg gagcggattt gaacgttgcg
      421 aagcaacggc ccggagggtg gcgggcagga cgcccgccat aaactgccag gcatcaaatt
      481 aagcagaagg ccatcctgac ggatggcctt tttgcgtttc tacaaactct tttgtttatt
      541 tttctaaata cattcaaata tgtatccgct catggatctc gatcccgcga aattaatacg
      601 actcactata gggtatttta actttactaa ggagcgatcg ctttcgaatt cctaccgcgg
      661 atatatggca gacattgaca acaaagaaca gtctgaactt gatcaagatt tggatgatgt
      721 tgaagaagta gaagaagagg aaactggtga agaaacaaaa ctcaaagcac gtcagctaac
      781 tgttcagatg atgcaaaatc ctcagattct tgcagccctt caagaaagac ttgatggtct
      841 ggtagaaaca ccaacaggat acattgaaag cctgcctagg gtagttaaaa gacgagtgaa
      901 tgctctcaaa aacctgcaag ttaaatgtgc acagatagaa gccaaattct atgaggaagt
      961 tcatgatctt gaaaggaagt atgctgttct ctatcagcct ctatttgata agcgatttga
     1021 aattattaat gcaatttatg aacctacgga agaagaatgt gaatggaaac cagatgaaga
     1081 agatgagatt tcggaggaat tgaaagaaaa ggccaagatt gaagatgaga aaaaggatga
     1141 agaaaaagaa gaccccaaag gaattcctga attttggtta actgttttta agaatgttga
     1201 cttgctcagt gatatggttc aggaacacga tgaacctatt ctgaagcact tgaaagatat
     1261 taaagtgaag ttctcagatg ctggccagcc tatgagtttt gtcttagaat ttcactttga
     1321 acccaatgaa tattttacaa atgaagtgct gacaaagaca tacaggatga ggtcagaacc
     1381 agatgattct gatccctttt cttttgatgg accagaaatt atgggttgta cagggtgcca
     1441 gatagattgg aaaaaaggaa agaatgtcac tttgaaaact attaagaaga agcagaaaca
     1501 caagggacgt gggacagttc gtactgtgac taaaacagtt tccaatgact ctttctttaa
     1561 cttttttgcc cctcctgaag ttcctgagag tggagatctg gatgatgatg ctgaagctat
     1621 ccttgctgca gacttcgaaa ttggtcactt tttacgtgag cgtataatcc caagatcagt
     1681 gttatatttt actggagaag ctattgaaga tgatgatgat gattatgatg aagaaggtga
     1741 agaagcggat gaggaagggg aagaagaagg agatgaggaa aatgatccag actatgaccc
     1801 aaagaaggat caaaacccag cagagtgcaa gcagcagggg ccaaccactg aggatctgta
     1861 ctttcagagc gataacgatg gatccgaaat cggtactggc tttccattcg acccccatta
     1921 tgtggaagtc ctgggcgagc gcatgcacta cgtcgatgtt ggtccgcgcg atggcacccc
     1981 tgtgctgttc ctgcacggta acccgacctc ctcctacgtg tggcgcaaca tcatcccgca
     2041 tgttgcaccg acccatcgct gcattgctcc agacctgatc ggtatgggca aatccgacaa
     2101 accagacctg ggttatttct tcgacgacca cgtccgcttc atggatgcct tcatcgaagc
     2161 cctgggtctg gaagaggtcg tcctggtcat tcacgactgg ggctccgctc tgggtttcca
     2221 ctgggccaag cgcaatccag agcgcgtcaa aggtattgca tttatggagt tcatccgccc
     2281 tatcccgacc tgggacgaat ggccagaatt tgcccgcgag accttccagg ccttccgcac
     2341 caccgacgtc ggccgcaagc tgatcatcga tcagaacgtt tttatcgagg gtacgctgcc
     2401 gatgggtgtc gtccgcccgc tgactgaagt cgagatggac cattaccgcg agccgttcct
     2461 gaatcctgtt gaccgcgagc cactgtggcg cttcccaaac gagctgccaa tcgccggtga
     2521 gccagcgaac atcgtcgcgc tggtcgaaga atacatggac tggctgcacc agtcccctgt
     2581 cccgaagctg ctgttctggg gcaccccagg cgttctgatc ccaccggccg aagccgctcg
     2641 cctggccaaa agcctgccta actgcaaggc tgtggacatc ggcccgggtc tgaatctgct
     2701 gcaagaagac aacccggacc tgatcggcag cgagatcgcg cgctggctgt ctactctgga
     2761 gatttccggt catcatcacc atcaccacta atagtcaacc gctgagcaat aactagcata
     2821 accccttggg gcctctaaac gggtcttgag gggttttttg ctgaaaggag gaactatatc
     2881 cggttcgctt gctgtccata aaaccgccca gtctagctat cgccatgtaa gcccactgca
     2941 agctacctgc tttctctttg cgcttgcgtt ttcccttgtc cagatagccc agtagctgac
     3001 attcatccgg ggtcagcacc gtttctgcgg actggctttc tacgtaatgg tttcttagac
     3061 gtcaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat
     3121 acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg
     3181 aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc cttattccct tttttgcggc
     3241 attttgcctt cctgtttttg ctcacccaga aacgctggtg aaagtaaaag atgctgaaga
     3301 tcagttgggt gcacgagtgg gttacatcga actggatctc aacagcggta agatccttga
     3361 gagttttcgc cccgaagaac gttttccaat gatgagcact ttcaaagttc tgctatgtgg
     3421 cgcggtatta tcccgtattg acgccgggca agagcaactc ggtcgccgca tacactattc
     3481 tcagaatgac ttggttgagt actcaccagt cacagaaaag catcttacgg atggcatgac
     3541 agtaagagaa ttatgcagtg ctgccataac catgagtgat aacactgcgg ccaacttact
     3601 tctgacaact atcggaggac cgaaggagct aaccgctttt ttgcacaaca tgggggatca
     3661 tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa gccataccaa acgacgagcg
     3721 tgacaccacg atgcctgtag caatggcaac aacgttgcgc aaactattaa ctggcgaact
     3781 acttactcta gcttcccggc aacaattaat agactggatg gaggcggata aagttgcagg
     3841 accacttctg cgctcggccc ttccggctgg ctggtttatt gctgataaat ctggagccgg
     3901 tgagcgtggg tctcgcggta tcattgcagc actggggcca gatggtaagc cctcccgtat
     3961 cgtagttatc tacacgacgg ggagtcaggc aactatggat gaacgaaata gacagatcgc
     4021 tgagataggt gcctcactga ttaagcattg gtaattcgaa atgaccgacc aagcgacgcc
     4081 caaccggtat cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac
     4141 gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg
     4201 ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca
     4261 agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc
     4321 tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc
     4381 ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag
     4441 gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc
     4501 ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca
     4561 gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac a
//
