LOCUS       sequence-345520-        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_I474M-Halo
FEATURES             Location/Qualifiers
     source          1..5175
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      28..55
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     primer_bind     137..156
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        137..155
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             1957..1977
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="EDLYFQS"
     CDS             1993..2877
                     /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             2878..2895
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(2906..2924)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      2920..2967
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        3196..3300
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3301..3369,3370..4161)
                     /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(3519..3538)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      complement(4319..4907)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4399..4418)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      5025..5111
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
ORIGIN
        1 cataaactgc caggcatcaa attaagcaga aggccatcct gacggatggc ctttttgcgt
       61 ttctacaaac tcttttgttt atttttctaa atacattcaa atatgtatcc gctcatggat
      121 ctcgatcccg cgaaattaat acgactcact atagggtatt ttaactttac taaggagcga
      181 tcgcttccga attcctaccg cggatacatg aacaaactgt atatcggaaa cctcagcgag
      241 aacgccgccc cctcggacct agaaagtatc ttcaaggacg ccaagatccc ggtgtcggga
      301 cccttcctgg tgaagactgg ctacgcgttc gtggactgcc cggacgagag ctgggccctc
      361 aaggccatcg aggcgctttc aggtaaaata gaactgcacg ggaaacccat agaagttgag
      421 cactcggtcc caaaaaggca aaggattcgg aaacttcaga tacgaaatat cccgcctcat
      481 ttacagtggg aggtgctgga tagtttacta gtccagtatg gagtggtgga gagctgtgag
      541 caagtgaaca ctgactcgga aactgcagtt gtaaatgtaa cctattccag taaggaccaa
      601 gctagacaag cactagacaa actgaatgga tttcagttag agaatttcac cttgaaagta
      661 gcctatatcc ctgatgaaat ggccgcccag caaaacccct tgcagcagcc ccgaggtcgc
      721 cgggggcttg ggcagagggg ctcctcaagg caggggtctc caggatccgt atccaagcag
      781 aaaccatgtg atttgcctct gcgcctgctg gttcccaccc aatttgttgg agccatcata
      841 ggaaaagaag gtgccaccat tcggaacatc accaaacaga cccagtctaa aatcgatgtc
      901 caccgtaaag aaaatgcggg ggctgctgag aagtcgatta ctatcctctc tactcctgaa
      961 ggcacctctg cggcttgtaa gtctattctg gagattatgc ataaggaagc tcaagatata
     1021 aaattcacag aagagatccc cttgaagatt ttagctcata ataactttgt tggacgtctt
     1081 attggtaaag aaggaagaaa tcttaaaaaa attgagcaag acacagacac taaaatcacg
     1141 atatctccat tgcaggaatt gacgctgtat aatccagaac gcactattac agttaaaggc
     1201 aatgttgaga catgtgccaa agctgaggag gagatcatga agaaaatcag ggagtcttat
     1261 gaaaatgata ttgcttctat gaatcttcaa gcacatttaa ttcctggatt aaatctgaac
     1321 gccttgggtc tgttcccacc cacttcaggg atgccacctc ccacctcagg gcccccttca
     1381 gccatgactc ctccctaccc gcagtttgag caatcagaaa cggagactgt tcatctgttt
     1441 atcccagctc tatcagtcgg tgccatcatc ggcaagcagg gccagcacat caagcagctt
     1501 tctcgctttg ctggagcttc aattaagatt gctccagcgg aagcaccaga tgctaaagtg
     1561 aggatggtga ttatcactgg accaccagag gctcagttca aggctcaggg aagaatttat
     1621 ggaaaaatga aagaagaaaa ctttgttagt cctaaagaag aggtgaaact tgaagctcat
     1681 atcagagtgc catcctttgc tgctggcaga gttattggaa aaggaggcaa aacggtgaat
     1741 gaacttcaga atttgtcaag tgcagaagtt gttgtccctc gtgaccagac acctgatgag
     1801 aatgaccaag tggttgtcaa aataactggt cacttctatg cttgccaggt tgcccagaga
     1861 aaaattcagg aaattctgac tcaggtaaag cagcaccaac aacagaaggc tctgcaaagt
     1921 ggaccacctc agtcaagacg gaaggagcca accactgagg atctgtactt tcagagcgat
     1981 aacgatggat ccgaaatcgg tactggcttt ccattcgacc cccattatgt ggaagtcctg
     2041 ggcgagcgca tgcactacgt cgatgttggt ccgcgcgatg gcacccctgt gctgttcctg
     2101 cacggtaacc cgacctcctc ctacgtgtgg cgcaacatca tcccgcatgt tgcaccgacc
     2161 catcgctgca ttgctccaga cctgatcggt atgggcaaat ccgacaaacc agacctgggt
     2221 tatttcttcg acgaccacgt ccgcttcatg gatgccttca tcgaagccct gggtctggaa
     2281 gaggtcgtcc tggtcattca cgactggggc tccgctctgg gtttccactg ggccaagcgc
     2341 aatccagagc gcgtcaaagg tattgcattt atggagttca tccgccctat cccgacctgg
     2401 gacgaatggc cagaatttgc ccgcgagacc ttccaggcct tccgcaccac cgacgtcggc
     2461 cgcaagctga tcatcgatca gaacgttttt atcgagggta cgctgccgat gggtgtcgtc
     2521 cgcccgctga ctgaagtcga gatggaccat taccgcgagc cgttcctgaa tcctgttgac
     2581 cgcgagccac tgtggcgctt cccaaacgag ctgccaatcg ccggtgagcc agcgaacatc
     2641 gtcgcgctgg tcgaagaata catggactgg ctgcaccagt cccctgtccc gaagctgctg
     2701 ttctggggca ccccaggcgt tctgatccca ccggccgaag ccgctcgcct ggccaaaagc
     2761 ctgcctaact gcaaggctgt ggacatcggc ccgggtctga atctgctgca agaagacaac
     2821 ccggacctga tcggcagcga gatcgcgcgc tggctgtcta ctctggagat ttccggtcat
     2881 catcaccatc accactaata gtcaaccgct gagcaataac tagcataacc ccttggggcc
     2941 tctaaacggg tcttgagggg ttttttgctg aaaggaggaa ctatatccgg ttcgcttgct
     3001 gtccataaaa ccgcccagtc tagctatcgc catgtaagcc cactgcaagc tacctgcttt
     3061 ctctttgcgc ttgcgttttc ccttgtccag atagcccagt agctgacatt catccggggt
     3121 cagcaccgtt tctgcggact ggctttctac gtaatggttt cttagacgtc aggtggcact
     3181 tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg
     3241 tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt
     3301 atgagtattc aacatttccg tgtcgccctt attccctttt ttgcggcatt ttgccttcct
     3361 gtttttgctc acccagaaac gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca
     3421 cgagtgggtt acatcgaact ggatctcaac agcggtaaga tccttgagag ttttcgcccc
     3481 gaagaacgtt ttccaatgat gagcactttc aaagttctgc tatgtggcgc ggtattatcc
     3541 cgtattgacg ccgggcaaga gcaactcggt cgccgcatac actattctca gaatgacttg
     3601 gttgagtact caccagtcac agaaaagcat cttacggatg gcatgacagt aagagaatta
     3661 tgcagtgctg ccataaccat gagtgataac actgcggcca acttacttct gacaactatc
     3721 ggaggaccga aggagctaac cgcttttttg cacaacatgg gggatcatgt aactcgcctt
     3781 gatcgttggg aaccggagct gaatgaagcc ataccaaacg acgagcgtga caccacgatg
     3841 cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg gcgaactact tactctagct
     3901 tcccggcaac aattaataga ctggatggag gcggataaag ttgcaggacc acttctgcgc
     3961 tcggcccttc cggctggctg gtttattgct gataaatctg gagccggtga gcgtgggtct
     4021 cgcggtatca ttgcagcact ggggccagat ggtaagccct cccgtatcgt agttatctac
     4081 acgacgggga gtcaggcaac tatggatgaa cgaaatagac agatcgctga gataggtgcc
     4141 tcactgatta agcattggta attcgaaatg accgaccaag cgacgcccaa ccggtatcag
     4201 ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca
     4261 tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt
     4321 tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc
     4381 gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct
     4441 ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg
     4501 tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca
     4561 agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact
     4621 atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta
     4681 acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta
     4741 actacggcta cactagaagg acagtatttg gtatctgcgc tctgctgaag ccagttacct
     4801 tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt
     4861 tttttgtttg caagcagcag attacgcgca gaaaaaaagg atttcaagaa gatcctttga
     4921 tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg catatgatgg
     4981 tagtgtgggg tctccccatg cgagagtagg gaactgccag gcatcaaata aaacgaaagg
     5041 ctcagtcgaa agactgggcc tttcgtttta tctgttgttt gtcggtgaac gctctcctga
     5101 gtaggacaaa tccgccggga gcggatttga acgttgcgaa gcaacggccc ggagggtggc
     5161 gggcaggacg cccgc
//
