LOCUS       sequence-344845-        5039 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 5039)
  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 5039)
  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_PTBP1-Halo
FEATURES             Location/Qualifiers
     source          1..5039
                     /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"
     regulatory      400..409
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2026..2046
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="EDLYFQS"
     CDS             2062..2946
                     /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             2947..2964
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(2975..2993)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      2989..3036
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        3265..3369
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3370..3438,3439..4230)
                     /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(3588..3607)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      complement(4388..4976)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4468..4487)
                     /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 tccgaattcg ccaccatgga cggcattgtc
      421 ccagatatag ccgttggtac aaagcgggga tctgacgagc ttttctctac ttgtgtcact
      481 aacggaccgt ttatcatgag cagcaactcg gcttctgcag caaacggaaa tgacagcaag
      541 aagttcaaag gtgacagccg aagtgcaggc gtcccctcta gagtgatcca catccggaag
      601 ctccccatcg acgtcacgga gggggaagtc atctccctgg ggctgccctt tgggaaggtc
      661 accaacctcc tgatgctgaa ggggaaaaac caggccttca tcgagatgaa cacggaggag
      721 gctgccaaca ccatggtgaa ctactacacc tcggtgaccc ctgtgctgcg cggccagccc
      781 atctacatcc agttctccaa ccacaaggag ctgaagaccg acagctctcc caaccaggcg
      841 cgggcccagg cggccctgca ggcggtgaac tcggtccagt cggggaacct ggccttggct
      901 gcctcggcgg cggccgtgga cgcagggatg gcgatggccg ggcagagccc cgtgctcagg
      961 atcatcgtgg agaacctctt ctaccctgtg accctggatg tgctgcacca gattttctcc
     1021 aagttcggca cagtgttgaa gatcatcacc ttcaccaaga acaaccagtt ccaggccctg
     1081 ctgcagtatg cggaccccgt gagcgcccag cacgccaagc tgtcgctgga cgggcagaac
     1141 atctacaacg cctgctgcac gctgcgcatc gacttttcca agctcaccag cctcaacgtc
     1201 aagtacaaca atgacaagag ccgtgactac acacgcccag acctgccttc cggggacagc
     1261 cagccctcgc tggaccagac catggccgcg gccttcggcc tttccgttcc gaacgtccac
     1321 ggcgccctgg cccccctggc catcccctcg gcggcggcgg cagctgcggc ggcaggtcgg
     1381 atcgccatcc cgggcctggc gggggcagga aattctgtat tgctggtcag caacctcaac
     1441 ccagagagag tcacacccca aagcctcttt attcttttcg gcgtctacgg tgacgtgcag
     1501 cgcgtgaaga tcctgttcaa taagaaggag aacgccctag tgcagatggc ggacggcaac
     1561 caggcccagc tggccatgag ccacctgaac gggcacaagc tgcacgggaa gcccatccgc
     1621 atcacgctct cgaagcacca gaacgtgcag ctgccccgcg agggccagga ggaccagggc
     1681 ctgaccaagg actacggcaa ctcacccctg caccgcttca agaagccggg ctccaagaac
     1741 ttccagaaca tattcccgcc ctcggccacg ctgcacctct ccaacatccc gccctcagtc
     1801 tccgaggagg atctcaaggt cctgttttcc agcaatgggg gcgtcgtcaa aggattcaag
     1861 ttcttccaga aggaccgcaa gatggcactg atccagatgg gctccgtgga ggaggcggtc
     1921 caggccctca ttgacctgca caaccacgac ctcggggaga accaccacct gcgggtctcc
     1981 ttctccaagt ccaccatcgg cggctccggc ctcgagccaa ccactgagga tctgtacttt
     2041 cagagcgata acgatggatc cgaaatcggt actggctttc cattcgaccc ccattatgtg
     2101 gaagtcctgg gcgagcgcat gcactacgtc gatgttggtc cgcgcgatgg cacccctgtg
     2161 ctgttcctgc acggtaaccc gacctcctcc tacgtgtggc gcaacatcat cccgcatgtt
     2221 gcaccgaccc atcgctgcat tgctccagac ctgatcggta tgggcaaatc cgacaaacca
     2281 gacctgggtt atttcttcga cgaccacgtc cgcttcatgg atgccttcat cgaagccctg
     2341 ggtctggaag aggtcgtcct ggtcattcac gactggggct ccgctctggg tttccactgg
     2401 gccaagcgca atccagagcg cgtcaaaggt attgcattta tggagttcat ccgccctatc
     2461 ccgacctggg acgaatggcc agaatttgcc cgcgagacct tccaggcctt ccgcaccacc
     2521 gacgtcggcc gcaagctgat catcgatcag aacgttttta tcgagggtac gctgccgatg
     2581 ggtgtcgtcc gcccgctgac tgaagtcgag atggaccatt accgcgagcc gttcctgaat
     2641 cctgttgacc gcgagccact gtggcgcttc ccaaacgagc tgccaatcgc cggtgagcca
     2701 gcgaacatcg tcgcgctggt cgaagaatac atggactggc tgcaccagtc ccctgtcccg
     2761 aagctgctgt tctggggcac cccaggcgtt ctgatcccac cggccgaagc cgctcgcctg
     2821 gccaaaagcc tgcctaactg caaggctgtg gacatcggcc cgggtctgaa tctgctgcaa
     2881 gaagacaacc cggacctgat cggcagcgag atcgcgcgct ggctgtctac tctggagatt
     2941 tccggtcatc atcaccatca ccactaatag tcaaccgctg agcaataact agcataaccc
     3001 cttggggcct ctaaacgggt cttgaggggt tttttgctga aaggaggaac tatatccggt
     3061 tcgcttgctg tccataaaac cgcccagtct agctatcgcc atgtaagccc actgcaagct
     3121 acctgctttc tctttgcgct tgcgttttcc cttgtccaga tagcccagta gctgacattc
     3181 atccggggtc agcaccgttt ctgcggactg gctttctacg taatggtttc ttagacgtca
     3241 ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat
     3301 tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa
     3361 aggaagagta tgagtattca acatttccgt gtcgccctta ttcccttttt tgcggcattt
     3421 tgccttcctg tttttgctca cccagaaacg ctggtgaaag taaaagatgc tgaagatcag
     3481 ttgggtgcac gagtgggtta catcgaactg gatctcaaca gcggtaagat ccttgagagt
     3541 tttcgccccg aagaacgttt tccaatgatg agcactttca aagttctgct atgtggcgcg
     3601 gtattatccc gtattgacgc cgggcaagag caactcggtc gccgcataca ctattctcag
     3661 aatgacttgg ttgagtactc accagtcaca gaaaagcatc ttacggatgg catgacagta
     3721 agagaattat gcagtgctgc cataaccatg agtgataaca ctgcggccaa cttacttctg
     3781 acaactatcg gaggaccgaa ggagctaacc gcttttttgc acaacatggg ggatcatgta
     3841 actcgccttg atcgttggga accggagctg aatgaagcca taccaaacga cgagcgtgac
     3901 accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg cgaactactt
     3961 actctagctt cccggcaaca attaatagac tggatggagg cggataaagt tgcaggacca
     4021 cttctgcgct cggcccttcc ggctggctgg tttattgctg ataaatctgg agccggtgag
     4081 cgtgggtctc gcggtatcat tgcagcactg gggccagatg gtaagccctc ccgtatcgta
     4141 gttatctaca cgacggggag tcaggcaact atggatgaac gaaatagaca gatcgctgag
     4201 ataggtgcct cactgattaa gcattggtaa ttcgaaatga ccgaccaagc gacgcccaac
     4261 cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag
     4321 gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc
     4381 tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc
     4441 agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc
     4501 tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt
     4561 cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg
     4621 ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat
     4681 ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag
     4741 ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt
     4801 ggtggcctaa ctacggctac actagaagga cagtatttgg tatctgcgct ctgctgaagc
     4861 cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta
     4921 gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tttcaagaag
     4981 atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggg
//
