LOCUS       sequence-345511-        5213 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 5213)
  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 5213)
  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_IGF2BP1_R167C-Halo
FEATURES             Location/Qualifiers
     source          1..5213
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      48..75
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     primer_bind     157..176
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        157..175
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      233..242
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2015..2035
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="EDLYFQS"
     CDS             2051..2935
                     /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             2936..2953
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(2964..2982)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      2978..3025
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        3254..3358
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3359..3427,3428..4219)
                     /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(3577..3596)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      complement(4377..4965)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4457..4476)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      5083..5169
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
ORIGIN
        1 gtggcgggca ggacgcccgc cataaactgc caggcatcaa attaagcaga aggccatcct
       61 gacggatggc ctttttgcgt ttctacaaac tcttttgttt atttttctaa atacattcaa
      121 atatgtatcc gctcatggat ctcgatcccg cgaaattaat acgactcact atagggtatt
      181 ttaactttac taaggagcga tcgcttccga attcctaccg cggatatcta gagccaccat
      241 ggcgaacaag ctttacatcg gcaacctcaa cgagagcgtg acccccgcgg acttggagaa
      301 agtgtttgcg gagcacaaga tctcctacag cggccagttc ttggtcaaat ccggctacgc
      361 cttcgtggac tgcccggacg agcactgggc gatgaaggcc atcgaaactt tctccgggaa
      421 agtagaatta caaggaaaac gcttagagat tgaacattcg gtgcccaaaa aacaaaggag
      481 ccggaaaatt caaatccgaa atattccacc ccagctccga tgggaagtac tggacagcct
      541 gctggctcag tatggtacag tagagaactg tgagcaagtg aacaccgaga gtgagacggc
      601 agtggtgaat gtcacctatt ccaaccggga gcagaccagg caagccatca tgaagctgaa
      661 tggccaccag ttggagaacc atgccctgaa ggtctcctac atccccgatg agcagatagc
      721 acagggacct gagaatgggt gccgaggggg ctttggctct cggggtcagc cccgccaggg
      781 ctcacctgtg gcagcggggg ccccagccaa gcagcagcaa gtggacatcc cccttcggct
      841 cctggtgccc acccagtatg tgggtgccat tattggcaag gagggggcca ccatccgcaa
      901 catcacaaaa cagacccagt ccaagataga cgtgcatagg aaggagaacg caggtgcagc
      961 tgaaaaagcc atcagtgtgc actccacccc tgagggctgc tcctccgctt gtaagatgat
     1021 cttggagatt atgcataaag aggctaagga caccaaaacg gctgacgagg ttcccctgaa
     1081 gatcctggcc cataataact ttgtagggcg tctcattggc aaggaaggac ggaacctgaa
     1141 gaaggtagag caagataccg agacaaaaat caccatctcc tcgttgcaag accttaccct
     1201 ttacaaccct gagaggacca tcactgtgaa gggggccatc gagaattgtt gcagggccga
     1261 gcaggaaata atgaagaaag ttcgggaggc ctatgagaat gatgtggctg ccatgagcct
     1321 gcagtctcac ctgatccctg gcctgaacct ggctgctgta ggtcttttcc cagcttcatc
     1381 cagcgcagtc ccgccgcctc ccagcagcgt tactggggct gctccctata gctcctttat
     1441 gcaggctccc gagcaggaga tggtgcaggt gtttatcccc gcccaggcag tgggcgccat
     1501 catcggcaag aaggggcagc acatcaaaca gctctcccgg tttgccagcg cctccatcaa
     1561 gattgcacca cccgaaacac ctgactccaa agttcgtatg gttatcatca ctggaccgcc
     1621 agaggcccaa ttcaaggctc agggaagaat ctatggcaaa ctcaaggagg agaacttctt
     1681 tggtcccaag gaggaagtga agctggagac ccacatacgt gtgccagcat cagcagctgg
     1741 ccgggtcatt ggcaaaggtg gaaaaacggt gaacgagttg cagaatttga cggcagctga
     1801 ggtggtagta ccaagagacc agacccctga tgagaacgac caggtcatcg tgaaaatcat
     1861 cggacatttc tatgccagtc agatggctca acggaagatc cgagacatcc tggcccaggt
     1921 taagcagcag catcagaagg gacagagtaa ccaggcccag gcacggagga agggcggctc
     1981 cggctcctgc aggcgagctc tcgagccaac cactgaggat ctgtactttc agagcgataa
     2041 cgatggatcc gaaatcggta ctggctttcc attcgacccc cattatgtgg aagtcctggg
     2101 cgagcgcatg cactacgtcg atgttggtcc gcgcgatggc acccctgtgc tgttcctgca
     2161 cggtaacccg acctcctcct acgtgtggcg caacatcatc ccgcatgttg caccgaccca
     2221 tcgctgcatt gctccagacc tgatcggtat gggcaaatcc gacaaaccag acctgggtta
     2281 tttcttcgac gaccacgtcc gcttcatgga tgccttcatc gaagccctgg gtctggaaga
     2341 ggtcgtcctg gtcattcacg actggggctc cgctctgggt ttccactggg ccaagcgcaa
     2401 tccagagcgc gtcaaaggta ttgcatttat ggagttcatc cgccctatcc cgacctggga
     2461 cgaatggcca gaatttgccc gcgagacctt ccaggccttc cgcaccaccg acgtcggccg
     2521 caagctgatc atcgatcaga acgtttttat cgagggtacg ctgccgatgg gtgtcgtccg
     2581 cccgctgact gaagtcgaga tggaccatta ccgcgagccg ttcctgaatc ctgttgaccg
     2641 cgagccactg tggcgcttcc caaacgagct gccaatcgcc ggtgagccag cgaacatcgt
     2701 cgcgctggtc gaagaataca tggactggct gcaccagtcc cctgtcccga agctgctgtt
     2761 ctggggcacc ccaggcgttc tgatcccacc ggccgaagcc gctcgcctgg ccaaaagcct
     2821 gcctaactgc aaggctgtgg acatcggccc gggtctgaat ctgctgcaag aagacaaccc
     2881 ggacctgatc ggcagcgaga tcgcgcgctg gctgtctact ctggagattt ccggtcatca
     2941 tcaccatcac cactaatagt caaccgctga gcaataacta gcataacccc ttggggcctc
     3001 taaacgggtc ttgaggggtt ttttgctgaa aggaggaact atatccggtt cgcttgctgt
     3061 ccataaaacc gcccagtcta gctatcgcca tgtaagccca ctgcaagcta cctgctttct
     3121 ctttgcgctt gcgttttccc ttgtccagat agcccagtag ctgacattca tccggggtca
     3181 gcaccgtttc tgcggactgg ctttctacgt aatggtttct tagacgtcag gtggcacttt
     3241 tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta
     3301 tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtat
     3361 gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt gccttcctgt
     3421 ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg
     3481 agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt ttcgccccga
     3541 agaacgtttt ccaatgatga gcactttcaa agttctgcta tgtggcgcgg tattatcccg
     3601 tattgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga atgacttggt
     3661 tgagtactca ccagtcacag aaaagcatct tacggatggc atgacagtaa gagaattatg
     3721 cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga caactatcgg
     3781 aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa ctcgccttga
     3841 tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca ccacgatgcc
     3901 tgtagcaatg gcaacaacgt tgcgcaaact attaactggc gaactactta ctctagcttc
     3961 ccggcaacaa ttaatagact ggatggaggc ggataaagtt gcaggaccac ttctgcgctc
     4021 ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc gtgggtctcg
     4081 cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag ttatctacac
     4141 gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga taggtgcctc
     4201 actgattaag cattggtaat tcgaaatgac cgaccaagcg acgcccaacc ggtatcagct
     4261 cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg
     4321 tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc
     4381 cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga
     4441 aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct
     4501 cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg
     4561 gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag
     4621 ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat
     4681 cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac
     4741 aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac
     4801 tacggctaca ctagaaggac agtatttggt atctgcgctc tgctgaagcc agttaccttc
     4861 ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt
     4921 tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ttcaagaaga tcctttgatc
     4981 ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggca tatgatggta
     5041 gtgtggggtc tccccatgcg agagtaggga actgccaggc atcaaataaa acgaaaggct
     5101 cagtcgaaag actgggcctt tcgttttatc tgttgtttgt cggtgaacgc tctcctgagt
     5161 aggacaaatc cgccgggagc ggatttgaac gttgcgaagc aacggcccgg agg
//
