LOCUS       sequence-345585-        5475 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 5475)
  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 5475)
  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_HSPA9-Halo
FEATURES             Location/Qualifiers
     source          1..5475
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      31..58
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     primer_bind     140..159
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        140..158
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             2260..2280
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="EDLYFQS"
     CDS             2296..3180
                     /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             3181..3198
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(3209..3227)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      3223..3270
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        3499..3603
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3604..3672,3673..4464)
                     /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(3822..3841)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      complement(4622..5210)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4702..4721)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      5328..5414
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
ORIGIN
        1 cgccataaac tgccaggcat caaattaagc agaaggccat cctgacggat ggcctttttg
       61 cgtttctaca aactcttttg tttatttttc taaatacatt caaatatgta tccgctcatg
      121 gatctcgatc ccgcgaaatt aatacgactc actatagggt attttaactt tactaaggag
      181 cgatcgcttc cgaattccta ccgcggatat atgataagtg ccagccgagc tgcagcagcc
      241 cgtctcgtgg gcgccgcagc ctcccggggc cctacggccg cccgccacca ggatagctgg
      301 aatggcctta gtcatgaggc ttttagactt gtttcaaggc gggattatgc atcagaagca
      361 atcaagggag cagttgttgg tattgatttg ggtactacca actcctgcgt ggcagttatg
      421 gaaggtaaac aagcaaaggt gctggagaat gccgaaggtg ccagaaccac cccttcagtt
      481 gtggccttta cagcagatgg tgagcgactt gttggaatgc cggccaagcg acaggctgtc
      541 accaacccaa acaatacatt ttatgctacc aagcgtctca ttggccggcg atatgatgat
      601 cctgaagtac agaaagacat taaaaatgtt ccctttaaaa ttgtccgtgc ctccaatggt
      661 gatgcctggg ttgaggctca tgggaaattg tattctccga gtcagattgg agcatttgtg
      721 ttgatgaaga tgaaagagac tgcagaaaat tacttggggc acacagcaaa aaatgctgtg
      781 atcacagtcc cagcttattt caatgactcg cagagacagg ccactaaaga tgctggccag
      841 atatctggac tgaatgtgct tcgggtgatt aatgagccca cagctgctgc tcttgcctat
      901 ggtctagaca aatcagaaga caaagtcatt gctgtatatg atttaggtgg tggaactttt
      961 gatatttcta tcctggaaat tcagaaagga gtatttgagg tgaaatccac aaatggggat
     1021 accttcttag gtggggaaga ctttgaccag gccttgctac ggcacattgt gaaggagttc
     1081 aagagagaga caggggttga tttgactaaa gacaacatgg cacttcagag ggtacgggaa
     1141 gctgctgaaa aggctaaatg tgaactctcc tcatctgtgc agactgacat caatttgccc
     1201 tatcttacaa tggattcttc tggacccaag catttgaata tgaagttgac ccgtgctcaa
     1261 tttgaaggga ttgtcactga tctaatcaga aggactatcg ctccatgcca aaaagctatg
     1321 caagatgcag aagtcagcaa gagtgacata ggagaagtga ttcttgtggg tggcatgact
     1381 aggatgccca aggttcagca gactgtacag gatctttttg gcagagcccc aagtaaagct
     1441 gtcaatcctg atgaggctgt ggccattgga gctgccattc agggaggtgt gttggccggc
     1501 gatgtcacgg atgtgctgct ccttgatgtc actcccctgt ctctgggtat tgaaactcta
     1561 ggaggtgtct ttaccaaact tattaatagg aataccacta ttccaaccaa gaagagccag
     1621 gtattctcta ctgccgctga tggtcaaacg caagtggaaa ttaaagtgtg tcagggtgaa
     1681 agagagatgg ctggagacaa caaactcctt ggacagttta ctttgattgg aattccacca
     1741 gcccctcgtg gagttcctca gattgaagtt acatttgaca ttgatgccaa tgggatagta
     1801 catgtttctg ctaaagataa aggcacagga cgtgagcagc agattgtaat ccagtcttct
     1861 ggtggattaa gcaaagatga tattgaaaat atggttaaaa atgcagagaa atatgctgaa
     1921 gaagaccggc gaaagaagga acgagttgaa gcagttaata tggctgaagg aatcattcac
     1981 gacacagaaa ccaagatgga agaattcaag gaccaattac ctgctgatga gtgcaacaag
     2041 ctgaaagaag agatttccaa aatgagggag ctcctggcta gaaaagacag cgaaacagga
     2101 gaaaatatta gacaggcagc atcctctctt cagcaggcat cattgaagct gttcgaaatg
     2161 gcatacaaaa agatggcatc tgagcgagaa ggctctggaa gttctggcac tggggaacaa
     2221 aaggaagatc aaaaggagga aaaacaggag ccaaccactg aggatctgta ctttcagagc
     2281 gataacgatg gatccgaaat cggtactggc tttccattcg acccccatta tgtggaagtc
     2341 ctgggcgagc gcatgcacta cgtcgatgtt ggtccgcgcg atggcacccc tgtgctgttc
     2401 ctgcacggta acccgacctc ctcctacgtg tggcgcaaca tcatcccgca tgttgcaccg
     2461 acccatcgct gcattgctcc agacctgatc ggtatgggca aatccgacaa accagacctg
     2521 ggttatttct tcgacgacca cgtccgcttc atggatgcct tcatcgaagc cctgggtctg
     2581 gaagaggtcg tcctggtcat tcacgactgg ggctccgctc tgggtttcca ctgggccaag
     2641 cgcaatccag agcgcgtcaa aggtattgca tttatggagt tcatccgccc tatcccgacc
     2701 tgggacgaat ggccagaatt tgcccgcgag accttccagg ccttccgcac caccgacgtc
     2761 ggccgcaagc tgatcatcga tcagaacgtt tttatcgagg gtacgctgcc gatgggtgtc
     2821 gtccgcccgc tgactgaagt cgagatggac cattaccgcg agccgttcct gaatcctgtt
     2881 gaccgcgagc cactgtggcg cttcccaaac gagctgccaa tcgccggtga gccagcgaac
     2941 atcgtcgcgc tggtcgaaga atacatggac tggctgcacc agtcccctgt cccgaagctg
     3001 ctgttctggg gcaccccagg cgttctgatc ccaccggccg aagccgctcg cctggccaaa
     3061 agcctgccta actgcaaggc tgtggacatc ggcccgggtc tgaatctgct gcaagaagac
     3121 aacccggacc tgatcggcag cgagatcgcg cgctggctgt ctactctgga gatttccggt
     3181 catcatcacc atcaccacta atagtcaacc gctgagcaat aactagcata accccttggg
     3241 gcctctaaac gggtcttgag gggttttttg ctgaaaggag gaactatatc cggttcgctt
     3301 gctgtccata aaaccgccca gtctagctat cgccatgtaa gcccactgca agctacctgc
     3361 tttctctttg cgcttgcgtt ttcccttgtc cagatagccc agtagctgac attcatccgg
     3421 ggtcagcacc gtttctgcgg actggctttc tacgtaatgg tttcttagac gtcaggtggc
     3481 acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat
     3541 atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag
     3601 agtatgagta ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt
     3661 cctgtttttg ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt
     3721 gcacgagtgg gttacatcga actggatctc aacagcggta agatccttga gagttttcgc
     3781 cccgaagaac gttttccaat gatgagcact ttcaaagttc tgctatgtgg cgcggtatta
     3841 tcccgtattg acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac
     3901 ttggttgagt actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa
     3961 ttatgcagtg ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaact
     4021 atcggaggac cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc
     4081 cttgatcgtt gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg
     4141 atgcctgtag caatggcaac aacgttgcgc aaactattaa ctggcgaact acttactcta
     4201 gcttcccggc aacaattaat agactggatg gaggcggata aagttgcagg accacttctg
     4261 cgctcggccc ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggg
     4321 tctcgcggta tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc
     4381 tacacgacgg ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt
     4441 gcctcactga ttaagcattg gtaattcgaa atgaccgacc aagcgacgcc caaccggtat
     4501 cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga
     4561 acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt
     4621 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt
     4681 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc
     4741 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa
     4801 gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct
     4861 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta
     4921 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg
     4981 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc
     5041 ctaactacgg ctacactaga aggacagtat ttggtatctg cgctctgctg aagccagtta
     5101 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg
     5161 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatttcaa gaagatcctt
     5221 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggcatatga
     5281 tggtagtgtg gggtctcccc atgcgagagt agggaactgc caggcatcaa ataaaacgaa
     5341 aggctcagtc gaaagactgg gcctttcgtt ttatctgttg tttgtcggtg aacgctctcc
     5401 tgagtaggac aaatccgccg ggagcggatt tgaacgttgc gaagcaacgg cccggagggt
     5461 ggcgggcagg acgcc
//
