LOCUS       sequence-345591-        3999 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 3999)
  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 3999)
  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_PEBP1-Halo
FEATURES             Location/Qualifiers
     source          1..3999
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     15..34
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        15..33
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             659..679
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="EDLYFQS"
     CDS             695..1579
                     /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             1580..1597
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(1608..1626)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1622..1669
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        1898..2002
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2003..2071,2072..2863)
                     /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(2221..2240)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      complement(3021..3609)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3101..3120)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      3727..3813
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      3905..3932
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
ORIGIN
        1 cgatcccgcg aaattaatac gactcactat agggtatttt aactttacta aggagcgatc
       61 gcttccgaat tcctaccgcg gatatatgcc ggtggacctc agcaagtggt ccgggccctt
      121 gagcctgcaa gaagtggacg agcagccgca gcacccgctg catgtcacct acgccggggc
      181 ggcggtggac gagctgggca aagtgctgac gcccacccag gttaagaata gacccaccag
      241 catttcgtgg gatggtcttg attcagggaa gctctacacc ttggtcctga cagacccgga
      301 tgctcccagc aggaaggatc ccaaatacag agaatggcat catttcctgg tggtcaacat
      361 gaagggcaat gacatcagca gtggcacagt cctctccgat tatgtgggct cggggcctcc
      421 caagggcaca ggcctccacc gctatgtctg gctggtttac gagcaggaca ggccgctaaa
      481 gtgtgacgag cccatcctca gcaaccgatc tggagaccac cgtggcaaat tcaaggtggc
      541 gtccttccgt aaaaagtatg agctcagggc cccggtggct ggcacgtgtt accaggccga
      601 gtgggatgac tatgtgccca aactgtacga gcagctgtct gggaaggggc caaccactga
      661 ggatctgtac tttcagagcg ataacgatgg atccgaaatc ggtactggct ttccattcga
      721 cccccattat gtggaagtcc tgggcgagcg catgcactac gtcgatgttg gtccgcgcga
      781 tggcacccct gtgctgttcc tgcacggtaa cccgacctcc tcctacgtgt ggcgcaacat
      841 catcccgcat gttgcaccga cccatcgctg cattgctcca gacctgatcg gtatgggcaa
      901 atccgacaaa ccagacctgg gttatttctt cgacgaccac gtccgcttca tggatgcctt
      961 catcgaagcc ctgggtctgg aagaggtcgt cctggtcatt cacgactggg gctccgctct
     1021 gggtttccac tgggccaagc gcaatccaga gcgcgtcaaa ggtattgcat ttatggagtt
     1081 catccgccct atcccgacct gggacgaatg gccagaattt gcccgcgaga ccttccaggc
     1141 cttccgcacc accgacgtcg gccgcaagct gatcatcgat cagaacgttt ttatcgaggg
     1201 tacgctgccg atgggtgtcg tccgcccgct gactgaagtc gagatggacc attaccgcga
     1261 gccgttcctg aatcctgttg accgcgagcc actgtggcgc ttcccaaacg agctgccaat
     1321 cgccggtgag ccagcgaaca tcgtcgcgct ggtcgaagaa tacatggact ggctgcacca
     1381 gtcccctgtc ccgaagctgc tgttctgggg caccccaggc gttctgatcc caccggccga
     1441 agccgctcgc ctggccaaaa gcctgcctaa ctgcaaggct gtggacatcg gcccgggtct
     1501 gaatctgctg caagaagaca acccggacct gatcggcagc gagatcgcgc gctggctgtc
     1561 tactctggag atttccggtc atcatcacca tcaccactaa tagtcaaccg ctgagcaata
     1621 actagcataa ccccttgggg cctctaaacg ggtcttgagg ggttttttgc tgaaaggagg
     1681 aactatatcc ggttcgcttg ctgtccataa aaccgcccag tctagctatc gccatgtaag
     1741 cccactgcaa gctacctgct ttctctttgc gcttgcgttt tcccttgtcc agatagccca
     1801 gtagctgaca ttcatccggg gtcagcaccg tttctgcgga ctggctttct acgtaatggt
     1861 ttcttagacg tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt
     1921 tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca
     1981 ataatattga aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc ttattccctt
     2041 ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctggtga aagtaaaaga
     2101 tgctgaagat cagttgggtg cacgagtggg ttacatcgaa ctggatctca acagcggtaa
     2161 gatccttgag agttttcgcc ccgaagaacg ttttccaatg atgagcactt tcaaagttct
     2221 gctatgtggc gcggtattat cccgtattga cgccgggcaa gagcaactcg gtcgccgcat
     2281 acactattct cagaatgact tggttgagta ctcaccagtc acagaaaagc atcttacgga
     2341 tggcatgaca gtaagagaat tatgcagtgc tgccataacc atgagtgata acactgcggc
     2401 caacttactt ctgacaacta tcggaggacc gaaggagcta accgcttttt tgcacaacat
     2461 gggggatcat gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa
     2521 cgacgagcgt gacaccacga tgcctgtagc aatggcaaca acgttgcgca aactattaac
     2581 tggcgaacta cttactctag cttcccggca acaattaata gactggatgg aggcggataa
     2641 agttgcagga ccacttctgc gctcggccct tccggctggc tggtttattg ctgataaatc
     2701 tggagccggt gagcgtgggt ctcgcggtat cattgcagca ctggggccag atggtaagcc
     2761 ctcccgtatc gtagttatct acacgacggg gagtcaggca actatggatg aacgaaatag
     2821 acagatcgct gagataggtg cctcactgat taagcattgg taattcgaaa tgaccgacca
     2881 agcgacgccc aaccggtatc agctcactca aaggcggtaa tacggttatc cacagaatca
     2941 ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa
     3001 aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat
     3061 cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc
     3121 cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc
     3181 gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt
     3241 tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac
     3301 cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg
     3361 ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca
     3421 gagttcttga agtggtggcc taactacggc tacactagaa ggacagtatt tggtatctgc
     3481 gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa
     3541 accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa
     3601 ggatttcaag aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac
     3661 tcacgttaag ggcatatgat ggtagtgtgg ggtctcccca tgcgagagta gggaactgcc
     3721 aggcatcaaa taaaacgaaa ggctcagtcg aaagactggg cctttcgttt tatctgttgt
     3781 ttgtcggtga acgctctcct gagtaggaca aatccgccgg gagcggattt gaacgttgcg
     3841 aagcaacggc ccggagggtg gcgggcagga cgcccgccat aaactgccag gcatcaaatt
     3901 aagcagaagg ccatcctgac ggatggcctt tttgcgtttc tacaaactct tttgtttatt
     3961 tttctaaata cattcaaata tgtatccgct catggatct
//
