LOCUS       sequence-350743-        4740 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 4740)
  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 4740)
  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_ENO1-Halo
FEATURES             Location/Qualifiers
     source          1..4740
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      55..141
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      233..260
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     primer_bind     342..361
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        342..360
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             1727..1747
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="EDLYFQS"
     CDS             1763..2647
                     /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             2648..2665
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(2676..2694)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      2690..2737
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        2966..3070
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3071..3139,3140..3931)
                     /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(3289..3308)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      complement(4089..4677)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4169..4188)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 catatgatgg tagtgtgggg tctccccatg cgagagtagg gaactgccag gcatcaaata
       61 aaacgaaagg ctcagtcgaa agactgggcc tttcgtttta tctgttgttt gtcggtgaac
      121 gctctcctga gtaggacaaa tccgccggga gcggatttga acgttgcgaa gcaacggccc
      181 ggagggtggc gggcaggacg cccgccataa actgccaggc atcaaattaa gcagaaggcc
      241 atcctgacgg atggcctttt tgcgtttcta caaactcttt tgtttatttt tctaaataca
      301 ttcaaatatg tatccgctca tggatctcga tcccgcgaaa ttaatacgac tcactatagg
      361 gtattttaac tttactaagg agcgatcgct tccgaattcc taccgcggat atatgtctat
      421 tctcaagatc catgccaggg agatctttga ctctcgcggg aatcccactg ttgaggttga
      481 tctcttcacc tcaaaaggtc tcttcagagc tgctgtgccc agtggtgctt caactggtat
      541 ctatgaggcc ctagagctcc gggacaatga taagactcgc tatatgggga agggtgtctc
      601 aaaggctgtt gagcacatca ataaaactat tgcgcctgcc ctggttagca agaaactgaa
      661 cgtcacagaa caagagaaga ttgacaaact gatgatcgag atggatggaa cagaaaataa
      721 atctaagttt ggtgcgaacg ccattctggg ggtgtccctt gccgtctgca aagctggtgc
      781 cgttgagaag ggggtccccc tgtaccgcca catcgctgac ttggctggca actctgaagt
      841 catcctgcca gtcccggcgt tcaatgtcat caatggcggt tctcatgctg gcaacaagct
      901 ggccatgcag gagttcatga tcctcccagt cggtgcagca aacttcaggg aagccatgcg
      961 cattggagca gaggtttacc acaacctgaa gaatgtcatc aaggagaaat atgggaaaga
     1021 tgccaccaat gtgggggatg aaggcgggtt tgctcccaac atcctggaga ataaagaagg
     1081 cctggagctg ctgaagactg ctattgggaa agctggctac actgataagg tggtcatcgg
     1141 catggacgta gcggcctccg agttcttcag gtctgggaag tatgacctgg acttcaagtc
     1201 tcccgatgac cccagcaggt acatctcgcc tgaccagctg gctgacctgt acaagtcctt
     1261 catcaaggac tacccagtgg tgtctatcga agatcccttt gaccaggatg actggggagc
     1321 ttggcagaag ttcacagcca gtgcaggaat ccaggtagtg ggggatgatc tcacagtgac
     1381 caacccaaag aggatcgcca aggccgtgaa cgagaagtcc tgcaactgcc tcctgctcaa
     1441 agtcaaccag attggctccg tgaccgagtc tcttcaggcg tgcaagctgg cccaggccaa
     1501 tggttggggc gtcatggtgt ctcatcgttc gggggagact gaagatacct tcatcgctga
     1561 cctggttgtg gggctgtgca ctgggcagat caagactggt gccccttgcc gatctgagcg
     1621 cttggccaag tacaaccagc tcctcagaat tgaagaggag ctgggcagca aggctaagtt
     1681 tgccggcagg aacttcagaa accccttggc caaggrgcca accactgagg atctgtactt
     1741 tcagagcgat aacgatggat ccgaaatcgg tactggcttt ccattcgacc cccattatgt
     1801 ggaagtcctg ggcgagcgca tgcactacgt cgatgttggt ccgcgcgatg gcacccctgt
     1861 gctgttcctg cacggtaacc cgacctcctc ctacgtgtgg cgcaacatca tcccgcatgt
     1921 tgcaccgacc catcgctgca ttgctccaga cctgatcggt atgggcaaat ccgacaaacc
     1981 agacctgggt tatttcttcg acgaccacgt ccgcttcatg gatgccttca tcgaagccct
     2041 gggtctggaa gaggtcgtcc tggtcattca cgactggggc tccgctctgg gtttccactg
     2101 ggccaagcgc aatccagagc gcgtcaaagg tattgcattt atggagttca tccgccctat
     2161 cccgacctgg gacgaatggc cagaatttgc ccgcgagacc ttccaggcct tccgcaccac
     2221 cgacgtcggc cgcaagctga tcatcgatca gaacgttttt atcgagggta cgctgccgat
     2281 gggtgtcgtc cgcccgctga ctgaagtcga gatggaccat taccgcgagc cgttcctgaa
     2341 tcctgttgac cgcgagccac tgtggcgctt cccaaacgag ctgccaatcg ccggtgagcc
     2401 agcgaacatc gtcgcgctgg tcgaagaata catggactgg ctgcaccagt cccctgtccc
     2461 gaagctgctg ttctggggca ccccaggcgt tctgatccca ccggccgaag ccgctcgcct
     2521 ggccaaaagc ctgcctaact gcaaggctgt ggacatcggc ccgggtctga atctgctgca
     2581 agaagacaac ccggacctga tcggcagcga gatcgcgcgc tggctgtcta ctctggagat
     2641 ttccggtcat catcaccatc accactaata gtcaaccgct gagcaataac tagcataacc
     2701 ccttggggcc tctaaacggg tcttgagggg ttttttgctg aaaggaggaa ctatatccgg
     2761 ttcgcttgct gtccataaaa ccgcccagtc tagctatcgc catgtaagcc cactgcaagc
     2821 tacctgcttt ctctttgcgc ttgcgttttc ccttgtccag atagcccagt agctgacatt
     2881 catccggggt cagcaccgtt tctgcggact ggctttctac gtaatggttt cttagacgtc
     2941 aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca
     3001 ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa
     3061 aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt ttgcggcatt
     3121 ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg ctgaagatca
     3181 gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga tccttgagag
     3241 ttttcgcccc gaagaacgtt ttccaatgat gagcactttc aaagttctgc tatgtggcgc
     3301 ggtattatcc cgtattgacg ccgggcaaga gcaactcggt cgccgcatac actattctca
     3361 gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg gcatgacagt
     3421 aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca acttacttct
     3481 gacaactatc ggaggaccga aggagctaac cgcttttttg cacaacatgg gggatcatgt
     3541 aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg acgagcgtga
     3601 caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg gcgaactact
     3661 tactctagct tcccggcaac aattaataga ctggatggag gcggataaag ttgcaggacc
     3721 acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg gagccggtga
     3781 gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct cccgtatcgt
     3841 agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac agatcgctga
     3901 gataggtgcc tcactgatta agcattggta attcgaaatg accgaccaag cgacgcccaa
     3961 ccggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca
     4021 ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg
     4081 ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt
     4141 cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc
     4201 ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct
     4261 tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc
     4321 gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta
     4381 tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca
     4441 gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag
     4501 tggtggccta actacggcta cactagaagg acagtatttg gtatctgcgc tctgctgaag
     4561 ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt
     4621 agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg atttcaagaa
     4681 gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg
//
