LOCUS       sequence-345489-        4585 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 4585)
  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 4585)
  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_RBFOX2-Halo
FEATURES             Location/Qualifiers
     source          1..4585
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             207..227
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="EDLYFQS"
     CDS             243..1127
                     /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             1128..1145
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(1156..1174)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1170..1217
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        1446..1550
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1551..1619,1620..2411)
                     /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(1769..1788)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      complement(2569..3157)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2649..2668)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      3235..3321
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      3413..3440
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     primer_bind     3522..3541
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        3522..3540
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      3598..3607
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
ORIGIN
        1 caaccgcagc caccgctgct gcagccgctg cagccgctta cagtgacggt tatggcaggg
       61 tgtacacagc cgacccctac catgcccttg cccctgccgc tagctatgga gttggcgctg
      121 tggcgagttt ataccgaggt ggctacagcc gatttgcccc ctacggcggc tccggctcct
      181 gcaggcgagc tctcgagcca accactgagg atctgtactt tcagagcgat aacgatggat
      241 ccgaaatcgg tactggcttt ccattcgacc cccattatgt ggaagtcctg ggcgagcgca
      301 tgcactacgt cgatgttggt ccgcgcgatg gcacccctgt gctgttcctg cacggtaacc
      361 cgacctcctc ctacgtgtgg cgcaacatca tcccgcatgt tgcaccgacc catcgctgca
      421 ttgctccaga cctgatcggt atgggcaaat ccgacaaacc agacctgggt tatttcttcg
      481 acgaccacgt ccgcttcatg gatgccttca tcgaagccct gggtctggaa gaggtcgtcc
      541 tggtcattca cgactggggc tccgctctgg gtttccactg ggccaagcgc aatccagagc
      601 gcgtcaaagg tattgcattt atggagttca tccgccctat cccgacctgg gacgaatggc
      661 cagaatttgc ccgcgagacc ttccaggcct tccgcaccac cgacgtcggc cgcaagctga
      721 tcatcgatca gaacgttttt atcgagggta cgctgccgat gggtgtcgtc cgcccgctga
      781 ctgaagtcga gatggaccat taccgcgagc cgttcctgaa tcctgttgac cgcgagccac
      841 tgtggcgctt cccaaacgag ctgccaatcg ccggtgagcc agcgaacatc gtcgcgctgg
      901 tcgaagaata catggactgg ctgcaccagt cccctgtccc gaagctgctg ttctggggca
      961 ccccaggcgt tctgatccca ccggccgaag ccgctcgcct ggccaaaagc ctgcctaact
     1021 gcaaggctgt ggacatcggc ccgggtctga atctgctgca agaagacaac ccggacctga
     1081 tcggcagcga gatcgcgcgc tggctgtcta ctctggagat ttccggtcat catcaccatc
     1141 accactaata gtcaaccgct gagcaataac tagcataacc ccttggggcc tctaaacggg
     1201 tcttgagggg ttttttgctg aaaggaggaa ctatatccgg ttcgcttgct gtccataaaa
     1261 ccgcccagtc tagctatcgc catgtaagcc cactgcaagc tacctgcttt ctctttgcgc
     1321 ttgcgttttc ccttgtccag atagcccagt agctgacatt catccggggt cagcaccgtt
     1381 tctgcggact ggctttctac gtaatggttt cttagacgtc aggtggcact tttcggggaa
     1441 atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca
     1501 tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt atgagtattc
     1561 aacatttccg tgtcgccctt attccctttt ttgcggcatt ttgccttcct gtttttgctc
     1621 acccagaaac gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt
     1681 acatcgaact ggatctcaac agcggtaaga tccttgagag ttttcgcccc gaagaacgtt
     1741 ttccaatgat gagcactttc aaagttctgc tatgtggcgc ggtattatcc cgtattgacg
     1801 ccgggcaaga gcaactcggt cgccgcatac actattctca gaatgacttg gttgagtact
     1861 caccagtcac agaaaagcat cttacggatg gcatgacagt aagagaatta tgcagtgctg
     1921 ccataaccat gagtgataac actgcggcca acttacttct gacaactatc ggaggaccga
     1981 aggagctaac cgcttttttg cacaacatgg gggatcatgt aactcgcctt gatcgttggg
     2041 aaccggagct gaatgaagcc ataccaaacg acgagcgtga caccacgatg cctgtagcaa
     2101 tggcaacaac gttgcgcaaa ctattaactg gcgaactact tactctagct tcccggcaac
     2161 aattaataga ctggatggag gcggataaag ttgcaggacc acttctgcgc tcggcccttc
     2221 cggctggctg gtttattgct gataaatctg gagccggtga gcgtgggtct cgcggtatca
     2281 ttgcagcact ggggccagat ggtaagccct cccgtatcgt agttatctac acgacgggga
     2341 gtcaggcaac tatggatgaa cgaaatagac agatcgctga gataggtgcc tcactgatta
     2401 agcattggta attcgaaatg accgaccaag cgacgcccaa ccggtatcag ctcactcaaa
     2461 ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa
     2521 aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct
     2581 ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac
     2641 aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc
     2701 gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc
     2761 tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg
     2821 tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga
     2881 gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta acaggattag
     2941 cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta
     3001 cactagaagg acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag
     3061 agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg
     3121 caagcagcag attacgcgca gaaaaaaagg atttcaagaa gatcctttga tcttttctac
     3181 ggggtctgac gctcagtgga acgaaaactt cacgttaagg gcatagccag gcatcaaata
     3241 aaacgaaagg ctcagtcgaa agactgggcc tttcgtttta tctgttgttt gtcggtgaac
     3301 gctctcctga gtaggacaaa tccgccggga gcggatttga acgttgcgaa gcaacggccc
     3361 ggagggtggc gggcaggacg cccgccataa actgccaggc atcaaattaa gcagaaggcc
     3421 atcctgacgg atggcctttt tgcgtttcta caaactcttt tgtttatttt tctaaataca
     3481 ttcaaatatg tatccgctca tggatctcga tcccgcgaaa ttaatacgac tcactatagg
     3541 gtattttaac tttactaagg agcgatcgct tccgaattcc taccgcggat atctagagcc
     3601 accatggcgg agaaaaagaa aatggtaact cagggtaacc aggagccgac aacaactcct
     3661 gacgcaatgg ttcagccttt tactaccatc ccatttccac cacctccgca gaatggaatt
     3721 cccacagagt atggggtgcc acacactcaa gactatgccg gccagaccgg tgagcataac
     3781 ctgacactct acggaagtac gcaagcccac ggggagcaga gcagcaactc acccagcaca
     3841 caaaatggat ctcttacgca gacagaaggt ggagcacaga cagacggcca gcagtcacag
     3901 acacaaagta gtgaaaattc agagagtaaa tctaccccga aacggctgca tgtctctaat
     3961 attcctttcc gcttccggga ccctgacctc cggcagatgt ttgggcagtt tggcaaaatc
     4021 ctagatgtag aaataatctt taatgaacgt ggctctaagg gattcgggtt cgtaactttc
     4081 gagaatagtg ctgatgcaga cagggccagg gagaaattac acggcaccgt ggtagagggc
     4141 cgtaaaatcg aggtgaataa tgctacagca cgtgtaatga ccaataagaa gatggtcaca
     4201 ccatatgcaa atggttggaa attaagccca gtagttggag ctgtatatgg tccggagtta
     4261 tatgcagcat ccagctttca agcagatgtg tccctaggca atgatgcagc agtgccccta
     4321 tcaggaagag ggggtatcaa cacttacatt cctttaatca gtctcccttt agttcctggc
     4381 ttcccttacc ctactgcagc caccacggca gccgctttca gaggagccca tttgaggggc
     4441 agagggcgga cagtatatgg tgcagtccga gcggtacctc caacagccat ccccgcctat
     4501 ccaggtgtgg tttaccagga cggattttac ggtgctgacc tctatggtgg atatgcagcc
     4561 tacagatatg cacagcctgc tactg
//
