LOCUS       sequence-345492-        5175 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 5175)
  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 5175)
  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_IGF2BP3-Halo
FEATURES             Location/Qualifiers
     source          1..5175
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      33..60
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     primer_bind     142..161
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        142..160
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             1962..1982
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="EDLYFQS"
     CDS             1998..2882
                     /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             2883..2900
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(2911..2929)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      2925..2972
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        3201..3305
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3306..3374,3375..4166)
                     /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(3524..3543)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      complement(4324..4912)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4404..4423)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      5030..5116
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
ORIGIN
        1 cccgccataa actgccaggc atcaaattaa gcagaaggcc atcctgacgg atggcctttt
       61 tgcgtttcta caaactcttt tgtttatttt tctaaataca ttcaaatatg tatccgctca
      121 tggatctcga tcccgcgaaa ttaatacgac tcactatagg gtattttaac tttactaagg
      181 agcgatcgct tccgaattcc taccgcggat acatgaacaa actgtatatc ggaaacctca
      241 gcgagaacgc cgccccctcg gacctagaaa gtatcttcaa ggacgccaag atcccggtgt
      301 cgggaccctt cctggtgaag actggctacg cgttcgtgga ctgcccggac gagagctggg
      361 ccctcaaggc catcgaggcg ctttcaggta aaatagaact gcacgggaaa cccatagaag
      421 ttgagcactc ggtcccaaaa aggcaaagga ttcggaaact tcagatacga aatatcccgc
      481 ctcatttaca gtgggaggtg ctggatagtt tactagtcca gtatggagtg gtggagagct
      541 gtgagcaagt gaacactgac tcggaaactg cagttgtaaa tgtaacctat tccagtaagg
      601 accaagctag acaagcacta gacaaactga atggatttca gttagagaat ttcaccttga
      661 aagtagccta tatccctgat gaaatggccg cccagcaaaa ccccttgcag cagccccgag
      721 gtcgccgggg gcttgggcag aggggctcct caaggcaggg gtctccagga tccgtatcca
      781 agcagaaacc atgtgatttg cctctgcgcc tgctggttcc cacccaattt gttggagcca
      841 tcataggaaa agaaggtgcc accattcgga acatcaccaa acagacccag tctaaaatcg
      901 atgtccaccg taaagaaaat gcgggggctg ctgagaagtc gattactatc ctctctactc
      961 ctgaaggcac ctctgcggct tgtaagtcta ttctggagat tatgcataag gaagctcaag
     1021 atataaaatt cacagaagag atccccttga agattttagc tcataataac tttgttggac
     1081 gtcttattgg taaagaagga agaaatctta aaaaaattga gcaagacaca gacactaaaa
     1141 tcacgatatc tccattgcag gaattgacgc tgtataatcc agaacgcact attacagtta
     1201 aaggcaatgt tgagacatgt gccaaagctg aggaggagat catgaagaaa atcagggagt
     1261 cttatgaaaa tgatattgct tctatgaatc ttcaagcaca tttaattcct ggattaaatc
     1321 tgaacgcctt gggtctgttc ccacccactt cagggatgcc acctcccacc tcagggcccc
     1381 cttcagccat gactcctccc tacccgcagt ttgagcaatc agaaacggag actgttcatc
     1441 tgtttatccc agctctatca gtcggtgcca tcatcggcaa gcagggccag cacatcaagc
     1501 agctttctcg ctttgctgga gcttcaatta agattgctcc agcggaagca ccagatgcta
     1561 aagtgaggat ggtgattatc actggaccac cagaggctca gttcaaggct cagggaagaa
     1621 tttatggaaa aattaaagaa gaaaactttg ttagtcctaa agaagaggtg aaacttgaag
     1681 ctcatatcag agtgccatcc tttgctgctg gcagagttat tggaaaagga ggcaaaacgg
     1741 tgaatgaact tcagaatttg tcaagtgcag aagttgttgt ccctcgtgac cagacacctg
     1801 atgagaatga ccaagtggtt gtcaaaataa ctggtcactt ctatgcttgc caggttgccc
     1861 agagaaaaat tcaggaaatt ctgactcagg taaagcagca ccaacaacag aaggctctgc
     1921 aaagtggacc acctcagtca agacggaagg agccaaccac tgaggatctg tactttcaga
     1981 gcgataacga tggatccgaa atcggtactg gctttccatt cgacccccat tatgtggaag
     2041 tcctgggcga gcgcatgcac tacgtcgatg ttggtccgcg cgatggcacc cctgtgctgt
     2101 tcctgcacgg taacccgacc tcctcctacg tgtggcgcaa catcatcccg catgttgcac
     2161 cgacccatcg ctgcattgct ccagacctga tcggtatggg caaatccgac aaaccagacc
     2221 tgggttattt cttcgacgac cacgtccgct tcatggatgc cttcatcgaa gccctgggtc
     2281 tggaagaggt cgtcctggtc attcacgact ggggctccgc tctgggtttc cactgggcca
     2341 agcgcaatcc agagcgcgtc aaaggtattg catttatgga gttcatccgc cctatcccga
     2401 cctgggacga atggccagaa tttgcccgcg agaccttcca ggccttccgc accaccgacg
     2461 tcggccgcaa gctgatcatc gatcagaacg tttttatcga gggtacgctg ccgatgggtg
     2521 tcgtccgccc gctgactgaa gtcgagatgg accattaccg cgagccgttc ctgaatcctg
     2581 ttgaccgcga gccactgtgg cgcttcccaa acgagctgcc aatcgccggt gagccagcga
     2641 acatcgtcgc gctggtcgaa gaatacatgg actggctgca ccagtcccct gtcccgaagc
     2701 tgctgttctg gggcacccca ggcgttctga tcccaccggc cgaagccgct cgcctggcca
     2761 aaagcctgcc taactgcaag gctgtggaca tcggcccggg tctgaatctg ctgcaagaag
     2821 acaacccgga cctgatcggc agcgagatcg cgcgctggct gtctactctg gagatttccg
     2881 gtcatcatca ccatcaccac taatagtcaa ccgctgagca ataactagca taaccccttg
     2941 gggcctctaa acgggtcttg aggggttttt tgctgaaagg aggaactata tccggttcgc
     3001 ttgctgtcca taaaaccgcc cagtctagct atcgccatgt aagcccactg caagctacct
     3061 gctttctctt tgcgcttgcg ttttcccttg tccagatagc ccagtagctg acattcatcc
     3121 ggggtcagca ccgtttctgc ggactggctt tctacgtaat ggtttcttag acgtcaggtg
     3181 gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa
     3241 atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga
     3301 agagtatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc
     3361 ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg
     3421 gtgcacgagt gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc
     3481 gccccgaaga acgttttcca atgatgagca ctttcaaagt tctgctatgt ggcgcggtat
     3541 tatcccgtat tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg
     3601 acttggttga gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag
     3661 aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa
     3721 ctatcggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc
     3781 gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca
     3841 cgatgcctgt agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc
     3901 tagcttcccg gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc
     3961 tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg
     4021 ggtctcgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta
     4081 tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag
     4141 gtgcctcact gattaagcat tggtaattcg aaatgaccga ccaagcgacg cccaaccggt
     4201 atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa
     4261 gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc
     4321 gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag
     4381 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt
     4441 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg
     4501 aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg
     4561 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg
     4621 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac
     4681 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg
     4741 gcctaactac ggctacacta gaaggacagt atttggtatc tgcgctctgc tgaagccagt
     4801 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg
     4861 tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatttc aagaagatcc
     4921 tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggcatat
     4981 gatggtagtg tggggtctcc ccatgcgaga gtagggaact gccaggcatc aaataaaacg
     5041 aaaggctcag tcgaaagact gggcctttcg ttttatctgt tgtttgtcgg tgaacgctct
     5101 cctgagtagg acaaatccgc cgggagcgga tttgaacgtt gcgaagcaac ggcccggagg
     5161 gtggcgggca ggacg
//
