LOCUS       sequence-345526-        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_R525C-Halo
FEATURES             Location/Qualifiers
     source          1..5175
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      256..342
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      434..461
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     primer_bind     543..562
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        543..561
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             2363..2383
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="EDLYFQS"
     CDS             2399..3283
                     /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             3284..3301
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(3312..3330)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      3326..3373
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        3602..3706
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3707..3775,3776..4567)
                     /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(3925..3944)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      complement(join(4725..5175,1..138))
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4805..4824)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc
       61 ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc
      121 agaaaaaaag gatttcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg
      181 aacgaaaact cacgttaagg gcatatgatg gtagtgtggg gtctccccat gcgagagtag
      241 ggaactgcca ggcatcaaat aaaacgaaag gctcagtcga aagactgggc ctttcgtttt
      301 atctgttgtt tgtcggtgaa cgctctcctg agtaggacaa atccgccggg agcggatttg
      361 aacgttgcga agcaacggcc cggagggtgg cgggcaggac gcccgccata aactgccagg
      421 catcaaatta agcagaaggc catcctgacg gatggccttt ttgcgtttct acaaactctt
      481 ttgtttattt ttctaaatac attcaaatat gtatccgctc atggatctcg atcccgcgaa
      541 attaatacga ctcactatag ggtattttaa ctttactaag gagcgatcgc ttccgaattc
      601 ctaccgcgga tacatgaaca aactgtatat cggaaacctc agcgagaacg ccgccccctc
      661 ggacctagaa agtatcttca aggacgccaa gatcccggtg tcgggaccct tcctggtgaa
      721 gactggctac gcgttcgtgg actgcccgga cgagagctgg gccctcaagg ccatcgaggc
      781 gctttcaggt aaaatagaac tgcacgggaa acccatagaa gttgagcact cggtcccaaa
      841 aaggcaaagg attcggaaac ttcagatacg aaatatcccg cctcatttac agtgggaggt
      901 gctggatagt ttactagtcc agtatggagt ggtggagagc tgtgagcaag tgaacactga
      961 ctcggaaact gcagttgtaa atgtaaccta ttccagtaag gaccaagcta gacaagcact
     1021 agacaaactg aatggatttc agttagagaa tttcaccttg aaagtagcct atatccctga
     1081 tgaaatggcc gcccagcaaa accccttgca gcagccccga ggtcgccggg ggcttgggca
     1141 gaggggctcc tcaaggcagg ggtctccagg atccgtatcc aagcagaaac catgtgattt
     1201 gcctctgcgc ctgctggttc ccacccaatt tgttggagcc atcataggaa aagaaggtgc
     1261 caccattcgg aacatcacca aacagaccca gtctaaaatc gatgtccacc gtaaagaaaa
     1321 tgcgggggct gctgagaagt cgattactat cctctctact cctgaaggca cctctgcggc
     1381 ttgtaagtct attctggaga ttatgcataa ggaagctcaa gatataaaat tcacagaaga
     1441 gatccccttg aagattttag ctcataataa ctttgttgga cgtcttattg gtaaagaagg
     1501 aagaaatctt aaaaaaattg agcaagacac agacactaaa atcacgatat ctccattgca
     1561 ggaattgacg ctgtataatc cagaacgcac tattacagtt aaaggcaatg ttgagacatg
     1621 tgccaaagct gaggaggaga tcatgaagaa aatcagggag tcttatgaaa atgatattgc
     1681 ttctatgaat cttcaagcac atttaattcc tggattaaat ctgaacgcct tgggtctgtt
     1741 cccacccact tcagggatgc cacctcccac ctcagggccc ccttcagcca tgactcctcc
     1801 ctacccgcag tttgagcaat cagaaacgga gactgttcat ctgtttatcc cagctctatc
     1861 agtcggtgcc atcatcggca agcagggcca gcacatcaag cagctttctc gctttgctgg
     1921 agcttcaatt aagattgctc cagcggaagc accagatgct aaagtgagga tggtgattat
     1981 cactggacca ccagaggctc agttcaaggc tcagggaaga atttatggaa aaattaaaga
     2041 agaaaacttt gttagtccta aagaagaggt gaaacttgaa gctcatatca gagtgccatc
     2101 ctttgctgct ggcagagtta ttggaaaagg aggcaaaacg gtgaatgaac ttcagaattt
     2161 gtcaagtgca gaagttgttg tcccttgtga ccagacacct gatgagaatg accaagtggt
     2221 tgtcaaaata actggtcact tctatgcttg ccaggttgcc cagagaaaaa ttcaggaaat
     2281 tctgactcag gtaaagcagc accaacaaca gaaggctctg caaagtggac cacctcagtc
     2341 aagacggaag gagccaacca ctgaggatct gtactttcag agcgataacg atggatccga
     2401 aatcggtact ggctttccat tcgaccccca ttatgtggaa gtcctgggcg agcgcatgca
     2461 ctacgtcgat gttggtccgc gcgatggcac ccctgtgctg ttcctgcacg gtaacccgac
     2521 ctcctcctac gtgtggcgca acatcatccc gcatgttgca ccgacccatc gctgcattgc
     2581 tccagacctg atcggtatgg gcaaatccga caaaccagac ctgggttatt tcttcgacga
     2641 ccacgtccgc ttcatggatg ccttcatcga agccctgggt ctggaagagg tcgtcctggt
     2701 cattcacgac tggggctccg ctctgggttt ccactgggcc aagcgcaatc cagagcgcgt
     2761 caaaggtatt gcatttatgg agttcatccg ccctatcccg acctgggacg aatggccaga
     2821 atttgcccgc gagaccttcc aggccttccg caccaccgac gtcggccgca agctgatcat
     2881 cgatcagaac gtttttatcg agggtacgct gccgatgggt gtcgtccgcc cgctgactga
     2941 agtcgagatg gaccattacc gcgagccgtt cctgaatcct gttgaccgcg agccactgtg
     3001 gcgcttccca aacgagctgc caatcgccgg tgagccagcg aacatcgtcg cgctggtcga
     3061 agaatacatg gactggctgc accagtcccc tgtcccgaag ctgctgttct ggggcacccc
     3121 aggcgttctg atcccaccgg ccgaagccgc tcgcctggcc aaaagcctgc ctaactgcaa
     3181 ggctgtggac atcggcccgg gtctgaatct gctgcaagaa gacaacccgg acctgatcgg
     3241 cagcgagatc gcgcgctggc tgtctactct ggagatttcc ggtcatcatc accatcacca
     3301 ctaatagtca accgctgagc aataactagc ataacccctt ggggcctcta aacgggtctt
     3361 gaggggtttt ttgctgaaag gaggaactat atccggttcg cttgctgtcc ataaaaccgc
     3421 ccagtctagc tatcgccatg taagcccact gcaagctacc tgctttctct ttgcgcttgc
     3481 gttttccctt gtccagatag cccagtagct gacattcatc cggggtcagc accgtttctg
     3541 cggactggct ttctacgtaa tggtttctta gacgtcaggt ggcacttttc ggggaaatgt
     3601 gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag
     3661 acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtatga gtattcaaca
     3721 tttccgtgtc gcccttattc ccttttttgc ggcattttgc cttcctgttt ttgctcaccc
     3781 agaaacgctg gtgaaagtaa aagatgctga agatcagttg ggtgcacgag tgggttacat
     3841 cgaactggat ctcaacagcg gtaagatcct tgagagtttt cgccccgaag aacgttttcc
     3901 aatgatgagc actttcaaag ttctgctatg tggcgcggta ttatcccgta ttgacgccgg
     3961 gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat gacttggttg agtactcacc
     4021 agtcacagaa aagcatctta cggatggcat gacagtaaga gaattatgca gtgctgccat
     4081 aaccatgagt gataacactg cggccaactt acttctgaca actatcggag gaccgaagga
     4141 gctaaccgct tttttgcaca acatggggga tcatgtaact cgccttgatc gttgggaacc
     4201 ggagctgaat gaagccatac caaacgacga gcgtgacacc acgatgcctg tagcaatggc
     4261 aacaacgttg cgcaaactat taactggcga actacttact ctagcttccc ggcaacaatt
     4321 aatagactgg atggaggcgg ataaagttgc aggaccactt ctgcgctcgg cccttccggc
     4381 tggctggttt attgctgata aatctggagc cggtgagcgt gggtctcgcg gtatcattgc
     4441 agcactgggg ccagatggta agccctcccg tatcgtagtt atctacacga cggggagtca
     4501 ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac tgattaagca
     4561 ttggtaattc gaaatgaccg accaagcgac gcccaaccgg tatcagctca ctcaaaggcg
     4621 gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc
     4681 cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc
     4741 ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga
     4801 ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc
     4861 ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat
     4921 agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg
     4981 cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc
     5041 aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga
     5101 gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact
     5161 agaaggacag tattt
//
