LOCUS       sequence-345579-        5376 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 5376)
  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 5376)
  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_HSPA8-Halo
FEATURES             Location/Qualifiers
     source          1..5376
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      33..60
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     primer_bind     142..161
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        142..160
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             2163..2183
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="EDLYFQS"
     CDS             2199..3083
                     /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             3084..3101
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(3112..3130)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      3126..3173
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        3402..3506
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3507..3575,3576..4367)
                     /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(3725..3744)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      complement(4525..5113)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4605..4624)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      5231..5317
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
ORIGIN
        1 cccgccataa actgccaggc atcaaattaa gcagaaggcc atcctgacgg atggcctttt
       61 tgcgtttcta caaactcttt tgtttatttt tctaaataca ttcaaatatg tatccgctca
      121 tggatctcga tcccgcgaaa ttaatacgac tcactatagg gtattttaac tttactaagg
      181 agcgatcgct tccgaattcc taccgcggat atatgtccaa gggacctgca gttggtattg
      241 atcttggcac cacctactct tgtgtgggtg ttttccagca cggaaaagtc gagataattg
      301 ccaatgatca gggaaaccga accactccaa gctatgtcgc ctttacggac actgaacggt
      361 tgatcggtga tgccgcaaag aatcaagttg caatgaaccc caccaacaca gtttttgatg
      421 ccaaacgtct gattggacgc agatttgatg atgctgttgt ccagtctgat atgaaacatt
      481 ggccctttat ggtggtgaat gatgctggca ggcccaaggt ccaagtagaa tacaagggag
      541 agaccaaaag cttctatcca gaggaggtgt cttctatggt tctgacaaag atgaaggaaa
      601 ttgcagaagc ctaccttggg aagactgtta ccaatgctgt ggtcacagtg ccagcttact
      661 ttaatgactc tcagcgtcag gctaccaaag atgctggaac tattgctggt ctcaatgtac
      721 ttagaattat taatgagcca actgctgctg ctattgctta cggcttagac aaaaaggttg
      781 gagcagaaag aaacgtgctc atctttgacc tgggaggtgg cacttttgat gtgtcaatcc
      841 tcactattga ggatggaatc tttgaggtca agtctacagc tggagacacc cacttgggtg
      901 gagaagattt tgacaaccga atggtcaacc attttattgc tgagtttaag cgcaagcata
      961 agaaggacat cagtgagaac aagagagctg taagacgcct ccgtactgct tgtgaacgtg
     1021 ctaagcgtac cctctcttcc agcacccagg ccagtattga gatcgattct ctctatgaag
     1081 gaatcgactt ctatacctcc attacccgtg cccgatttga agaactgaat gctgacctgt
     1141 tccgtggcac cctggaccca gtagagaaag cccttcgaga tgccaaacta gacaagtcac
     1201 agattcatga tattgtcctg gttggtggtt ctactcgtat ccccaagatt cagaagcttc
     1261 tccaagactt cttcaatgga aaagaactga ataagagcat caaccctgat gaagctgttg
     1321 cttatggtgc agctgtccag gcagccatct tgtctggaga caagtctgag aatgttcaag
     1381 atttgctgct cttggatgtc actcctcttt cccttggtat tgaaactgct ggtggagtca
     1441 tgactgtcct catcaagcgt aataccacca ttcctaccaa gcagacacag accttcacta
     1501 cctattctga caaccagcct ggtgtgctta ttcaggttta tgaaggcgag cgtgccatga
     1561 caaaggataa caacctgctt ggcaagtttg aactcacagg catacctcct gcaccccgag
     1621 gtgttcctca gattgaagtc acttttgaca ttgatgccaa tggtatactc aatgtctctg
     1681 ctgtggacaa gagtacggga aaagagaaca agattactat cactaatgac aagggccgtt
     1741 tgagcaagga agacattgaa cgtatggtcc aggaagctga gaagtacaaa gctgaagatg
     1801 agaagcagag ggacaaggtg tcatccaaga attcacttga gtcctatgcc ttcaacatga
     1861 aagcaactgt tgaagatgag aaacttcaag gcaagattaa cgatgaggac aaacagaaga
     1921 ttctggacaa gtgtaatgaa attatcaact ggcttgataa gaatcagact gctgagaagg
     1981 aagaatttga acatcaacag aaagagctgg agaaagtttg caaccccatc atcaccaagc
     2041 tgtaccagag tgcaggaggc atgccaggag gaatgcctgg gggatttcct ggtggtggag
     2101 ctcctccctc tggtggtgct tcctcagggc ccaccattga agaggttgat gggccaacca
     2161 ctgaggatct gtactttcag agcgataacg atggatccga aatcggtact ggctttccat
     2221 tcgaccccca ttatgtggaa gtcctgggcg agcgcatgca ctacgtcgat gttggtccgc
     2281 gcgatggcac ccctgtgctg ttcctgcacg gtaacccgac ctcctcctac gtgtggcgca
     2341 acatcatccc gcatgttgca ccgacccatc gctgcattgc tccagacctg atcggtatgg
     2401 gcaaatccga caaaccagac ctgggttatt tcttcgacga ccacgtccgc ttcatggatg
     2461 ccttcatcga agccctgggt ctggaagagg tcgtcctggt cattcacgac tggggctccg
     2521 ctctgggttt ccactgggcc aagcgcaatc cagagcgcgt caaaggtatt gcatttatgg
     2581 agttcatccg ccctatcccg acctgggacg aatggccaga atttgcccgc gagaccttcc
     2641 aggccttccg caccaccgac gtcggccgca agctgatcat cgatcagaac gtttttatcg
     2701 agggtacgct gccgatgggt gtcgtccgcc cgctgactga agtcgagatg gaccattacc
     2761 gcgagccgtt cctgaatcct gttgaccgcg agccactgtg gcgcttccca aacgagctgc
     2821 caatcgccgg tgagccagcg aacatcgtcg cgctggtcga agaatacatg gactggctgc
     2881 accagtcccc tgtcccgaag ctgctgttct ggggcacccc aggcgttctg atcccaccgg
     2941 ccgaagccgc tcgcctggcc aaaagcctgc ctaactgcaa ggctgtggac atcggcccgg
     3001 gtctgaatct gctgcaagaa gacaacccgg acctgatcgg cagcgagatc gcgcgctggc
     3061 tgtctactct ggagatttcc ggtcatcatc accatcacca ctaatagtca accgctgagc
     3121 aataactagc ataacccctt ggggcctcta aacgggtctt gaggggtttt ttgctgaaag
     3181 gaggaactat atccggttcg cttgctgtcc ataaaaccgc ccagtctagc tatcgccatg
     3241 taagcccact gcaagctacc tgctttctct ttgcgcttgc gttttccctt gtccagatag
     3301 cccagtagct gacattcatc cggggtcagc accgtttctg cggactggct ttctacgtaa
     3361 tggtttctta gacgtcaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt
     3421 tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc
     3481 ttcaataata ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc gcccttattc
     3541 ccttttttgc ggcattttgc cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa
     3601 aagatgctga agatcagttg ggtgcacgag tgggttacat cgaactggat ctcaacagcg
     3661 gtaagatcct tgagagtttt cgccccgaag aacgttttcc aatgatgagc actttcaaag
     3721 ttctgctatg tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc
     3781 gcatacacta ttctcagaat gacttggttg agtactcacc agtcacagaa aagcatctta
     3841 cggatggcat gacagtaaga gaattatgca gtgctgccat aaccatgagt gataacactg
     3901 cggccaactt acttctgaca actatcggag gaccgaagga gctaaccgct tttttgcaca
     3961 acatggggga tcatgtaact cgccttgatc gttgggaacc ggagctgaat gaagccatac
     4021 caaacgacga gcgtgacacc acgatgcctg tagcaatggc aacaacgttg cgcaaactat
     4081 taactggcga actacttact ctagcttccc ggcaacaatt aatagactgg atggaggcgg
     4141 ataaagttgc aggaccactt ctgcgctcgg cccttccggc tggctggttt attgctgata
     4201 aatctggagc cggtgagcgt gggtctcgcg gtatcattgc agcactgggg ccagatggta
     4261 agccctcccg tatcgtagtt atctacacga cggggagtca ggcaactatg gatgaacgaa
     4321 atagacagat cgctgagata ggtgcctcac tgattaagca ttggtaattc gaaatgaccg
     4381 accaagcgac gcccaaccgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga
     4441 atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg
     4501 taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa
     4561 aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt
     4621 tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct
     4681 gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct
     4741 cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc
     4801 cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt
     4861 atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc
     4921 tacagagttc ttgaagtggt ggcctaacta cggctacact agaaggacag tatttggtat
     4981 ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa
     5041 acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa
     5101 aaaaggattt caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga
     5161 aaactcacgt taagggcata tgatggtagt gtggggtctc cccatgcgag agtagggaac
     5221 tgccaggcat caaataaaac gaaaggctca gtcgaaagac tgggcctttc gttttatctg
     5281 ttgtttgtcg gtgaacgctc tcctgagtag gacaaatccg ccgggagcgg atttgaacgt
     5341 tgcgaagcaa cggcccggag ggtggcgggc aggacg
//
