LOCUS       sequence-345076-        4785 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 4785)
  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 4785)
  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_NASP-Halo
FEATURES             Location/Qualifiers
     source          1..4785
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      286..372
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      464..491
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     primer_bind     573..592
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        573..591
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             2003..2023
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="EDLYFQS"
     CDS             2039..2923
                     /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             2924..2941
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(2952..2970)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      2966..3013
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        3242..3346
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3347..3415,3416..4207)
                     /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(3565..3584)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      complement(join(4365..4785,1..168))
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4445..4464)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 aactacggct acactagaag gacagtattt ggtatctgcg ctctgctgaa gccagttacc
       61 ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt
      121 ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatttcaaga agatcctttg
      181 atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg gcatatgatg
      241 gtagtgtggg gtctccccat gcgagagtag ggaactgcca ggcatcaaat aaaacgaaag
      301 gctcagtcga aagactgggc ctttcgtttt atctgttgtt tgtcggtgaa cgctctcctg
      361 agtaggacaa atccgccggg agcggatttg aacgttgcga agcaacggcc cggagggtgg
      421 cgggcaggac gcccgccata aactgccagg catcaaatta agcagaaggc catcctgacg
      481 gatggccttt ttgcgtttct acaaactctt ttgtttattt ttctaaatac attcaaatat
      541 gtatccgctc atggatctcg atcccgcgaa attaatacga ctcactatag ggtattttaa
      601 ctttactaag gagcgatcgc ttccgaattc ctaccgcgga tatatggcca tggagtccac
      661 agccactgcc gccgtcgccg cggagctggt ttctgccgac aaaattgaag atgttcctgc
      721 tccttctaca tctgcagata aagtggagag tctggatgtg gatagtgaag ctaagaaact
      781 attgggttta ggacagaaac atctggtgat gggggatatt ccagcagctg tcaatgcatt
      841 ccaggaagca gctagtcttt taggtaagaa gtatggagag acagctaatg agtgtggaga
      901 agccttcttt ttctatggga aatcacttct ggagttggca agaatggaga atggtgtgtt
      961 gggaaacgcc ttggaaggtg tgcatgtgga agaggaagaa ggagaaaaaa cagaagatga
     1021 atctctggta gaaaataatg ataacataga tgaaactgaa ggctcggaag aggatgataa
     1081 agaaaatgat aagactgaag aaatgccaaa tgattcagtc cttgaaaaca agtctcttca
     1141 agaaaatgag gaggaggaga ttgggaacct agagcttgcc tgggatatgc tggatttagc
     1201 aaagatcatt tttaaaaggc aagaaacaaa agaagcacag ctttatgctg cccaggcaca
     1261 tcttaaactc ggagaagtta gtgttgaatc tgaaaactat gtgcaagctg tggaggagtt
     1321 ccagtcctgc cttaacctgc aggaacagta cctggaagcc cacgaccgtc tccttgcaga
     1381 gacccactac cagctgggct tggcttatgg gtacaactct cagtatgatg aggcagtggc
     1441 acagttcagc aaatctattg aagtcattga gaacagaatg gctgtactaa acgagcaggt
     1501 gaaggaggct gaaggatcgt ctgctgaata caagaaagaa attgaggaac taaaggaact
     1561 gctacccgaa attagagaga agatagaaga tgcaaaggag tctcagcgta gtgggaatgt
     1621 agctgaactg gctctgaaag ctactctggt ggagagttct acttcaggtt tcactcctgg
     1681 tggaggaggc tcttcagtct ccatgattgc cagtagaaag ccaacagacg gtgcttcctc
     1741 atcaaattgt gtgactgata tttcccacct tgtcagaaag aagaggaaac cagaggaaga
     1801 gagtccccgg aaagatgatg caaagaaagc caaacaagag ccggaggtga acggaggcag
     1861 tggggatgct gtccccagtg gaaatgaagt ttcggaaaac atggaggagg aggctgagaa
     1921 tcaggctgaa agccgggcag cagtggaggg gacagtggag gctggagcta cagttgaaag
     1981 cactgcatgt gggccaacca ctgaggatct gtactttcag agcgataacg atggatccga
     2041 aatcggtact ggctttccat tcgaccccca ttatgtggaa gtcctgggcg agcgcatgca
     2101 ctacgtcgat gttggtccgc gcgatggcac ccctgtgctg ttcctgcacg gtaacccgac
     2161 ctcctcctac gtgtggcgca acatcatccc gcatgttgca ccgacccatc gctgcattgc
     2221 tccagacctg atcggtatgg gcaaatccga caaaccagac ctgggttatt tcttcgacga
     2281 ccacgtccgc ttcatggatg ccttcatcga agccctgggt ctggaagagg tcgtcctggt
     2341 cattcacgac tggggctccg ctctgggttt ccactgggcc aagcgcaatc cagagcgcgt
     2401 caaaggtatt gcatttatgg agttcatccg ccctatcccg acctgggacg aatggccaga
     2461 atttgcccgc gagaccttcc aggccttccg caccaccgac gtcggccgca agctgatcat
     2521 cgatcagaac gtttttatcg agggtacgct gccgatgggt gtcgtccgcc cgctgactga
     2581 agtcgagatg gaccattacc gcgagccgtt cctgaatcct gttgaccgcg agccactgtg
     2641 gcgcttccca aacgagctgc caatcgccgg tgagccagcg aacatcgtcg cgctggtcga
     2701 agaatacatg gactggctgc accagtcccc tgtcccgaag ctgctgttct ggggcacccc
     2761 aggcgttctg atcccaccgg ccgaagccgc tcgcctggcc aaaagcctgc ctaactgcaa
     2821 ggctgtggac atcggcccgg gtctgaatct gctgcaagaa gacaacccgg acctgatcgg
     2881 cagcgagatc gcgcgctggc tgtctactct ggagatttcc ggtcatcatc accatcacca
     2941 ctaatagtca accgctgagc aataactagc ataacccctt ggggcctcta aacgggtctt
     3001 gaggggtttt ttgctgaaag gaggaactat atccggttcg cttgctgtcc ataaaaccgc
     3061 ccagtctagc tatcgccatg taagcccact gcaagctacc tgctttctct ttgcgcttgc
     3121 gttttccctt gtccagatag cccagtagct gacattcatc cggggtcagc accgtttctg
     3181 cggactggct ttctacgtaa tggtttctta gacgtcaggt ggcacttttc ggggaaatgt
     3241 gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag
     3301 acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtatga gtattcaaca
     3361 tttccgtgtc gcccttattc ccttttttgc ggcattttgc cttcctgttt ttgctcaccc
     3421 agaaacgctg gtgaaagtaa aagatgctga agatcagttg ggtgcacgag tgggttacat
     3481 cgaactggat ctcaacagcg gtaagatcct tgagagtttt cgccccgaag aacgttttcc
     3541 aatgatgagc actttcaaag ttctgctatg tggcgcggta ttatcccgta ttgacgccgg
     3601 gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat gacttggttg agtactcacc
     3661 agtcacagaa aagcatctta cggatggcat gacagtaaga gaattatgca gtgctgccat
     3721 aaccatgagt gataacactg cggccaactt acttctgaca actatcggag gaccgaagga
     3781 gctaaccgct tttttgcaca acatggggga tcatgtaact cgccttgatc gttgggaacc
     3841 ggagctgaat gaagccatac caaacgacga gcgtgacacc acgatgcctg tagcaatggc
     3901 aacaacgttg cgcaaactat taactggcga actacttact ctagcttccc ggcaacaatt
     3961 aatagactgg atggaggcgg ataaagttgc aggaccactt ctgcgctcgg cccttccggc
     4021 tggctggttt attgctgata aatctggagc cggtgagcgt gggtctcgcg gtatcattgc
     4081 agcactgggg ccagatggta agccctcccg tatcgtagtt atctacacga cggggagtca
     4141 ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac tgattaagca
     4201 ttggtaattc gaaatgaccg accaagcgac gcccaaccgg tatcagctca ctcaaaggcg
     4261 gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc
     4321 cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc
     4381 ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga
     4441 ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc
     4501 ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat
     4561 agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg
     4621 cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc
     4681 aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga
     4741 gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcct
//
