LOCUS       sequence-345518-        5106 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 5106)
  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 5106)
  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_IGF2BP2b-Halo
FEATURES             Location/Qualifiers
     source          1..5106
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      44..71
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     primer_bind     153..172
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        153..171
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             1904..1924
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="EDLYFQS"
     CDS             1940..2824
                     /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             2825..2842
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(2853..2871)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      2867..2914
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        3143..3247
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3248..3316,3317..4108)
                     /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(3466..3485)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      complement(4266..4854)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4346..4365)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      4972..5058
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
ORIGIN
        1 cgggcaggac gcccgccata aactgccagg catcaaatta agcagaaggc catcctgacg
       61 gatggccttt ttgcgtttct acaaactctt ttgtttattt ttctaaatac attcaaatat
      121 gtatccgctc atggatctcg atcccgcgaa attaatacga ctcactatag ggtattttaa
      181 ctttactaag gagcgatcgc ttccgaattc ctaccgcgga tatatgatga acaagcttta
      241 catcgggaac ctgagccccg ccgtcaccgc cgacgacctc cggcagctct ttggggacag
      301 gaagctgccc ctggcgggac aggtcctgct gaagtccggc tacgccttcg tggactaccc
      361 cgaccagaac tgggccatcc gcgccatcga gaccctctcg ggtaaagtgg aattgcatgg
      421 gaaaatcatg gaagttgatt actcagtctc taaaaagcta aggagcagga aaattcagat
      481 tcgaaacatc cctcctcacc tgcagtggga ggtgttggat ggacttttgg ctcaatatgg
      541 gacagtggag aatgtggaac aagtcaacac agacacagaa accgccgttg tcaacgtcac
      601 atatgcaaca agagaagaag caaaaatagc catggagaag ctaagcgggc atcagtttga
      661 gaactactcc ttcaagattt cctacatccc ggatgaagag gtgagctccc cttcgccccc
      721 tcagcgagcc cagcgtgggg accactcttc ccgggagcaa ggccacgccc ctgggggcac
      781 ttctcaggcc agacagattg atttcccgct gcggatcctg gtccccaccc agtttgttgg
      841 tgccatcatc ggaaaggagg gcttgaccat aaagaacatc actaagcaga cccagtcccg
      901 ggtagatatc catagaaaag agaactctgg agctgcagag aagcctgtca ccatccatgc
      961 caccccagag gggacttctg aagcatgccg catgattctt gaaatcatgc agaaagaggc
     1021 agatgagacc aaactagccg aagagattcc tctgaaaatc ttggcacaca atggcttggt
     1081 tggaagactg attggaaaag aaggcagaaa tttgaagaaa attgaacatg aaacagggac
     1141 caagataaca atctcatctt tgcaggattt gagcatatac aacccggaaa gaaccatcac
     1201 tgtgaagggc acagttgagg cctgtgccag tgctgagata gagattatga agaagctgcg
     1261 tgaggccttt gaaaatgata tgctggctgt taacacccac tccggatact tctccagcct
     1321 gtacccccat caccagtttg gcccgttccc gcatcatcac tcttatccag agcaggagat
     1381 tgtgaatctc ttcatcccaa cccaggctgt gggcgccatc atcgggaaga agggggcaca
     1441 catcaaacag ctggcgagat tcgccggagc ctctatcaag attgcccctg cggaaggccc
     1501 agacgtcagc gaaaggatgg tcatcatcac cgggccaccg gaagcccagt tcaaggccca
     1561 gggacggatc tttgggaaac tgaaagagga aaacttcttt aaccccaaag aagaagtgaa
     1621 gctggaagcg catatcagag tgccctcttc cacagctggc cgggtgattg gcaaaggtgg
     1681 caagaccgtg aacgaactgc agaacttaac cagtgcagaa gtcatcgtgc ctcgtgacca
     1741 aacgccagat gaaaatgagg aagtgatcgt cagaattatc gggcacttct ttgctagcca
     1801 gactgcacag cgcaagatca gggaaattgt acaacaggtg aagcagcagg agcagaaata
     1861 ccctcaggga gtcgcctcac agcgcagcaa ggggccaacc actgaggatc tgtactttca
     1921 gagcgataac gatggatccg aaatcggtac tggctttcca ttcgaccccc attatgtgga
     1981 agtcctgggc gagcgcatgc actacgtcga tgttggtccg cgcgatggca cccctgtgct
     2041 gttcctgcac ggtaacccga cctcctccta cgtgtggcgc aacatcatcc cgcatgttgc
     2101 accgacccat cgctgcattg ctccagacct gatcggtatg ggcaaatccg acaaaccaga
     2161 cctgggttat ttcttcgacg accacgtccg cttcatggat gccttcatcg aagccctggg
     2221 tctggaagag gtcgtcctgg tcattcacga ctggggctcc gctctgggtt tccactgggc
     2281 caagcgcaat ccagagcgcg tcaaaggtat tgcatttatg gagttcatcc gccctatccc
     2341 gacctgggac gaatggccag aatttgcccg cgagaccttc caggccttcc gcaccaccga
     2401 cgtcggccgc aagctgatca tcgatcagaa cgtttttatc gagggtacgc tgccgatggg
     2461 tgtcgtccgc ccgctgactg aagtcgagat ggaccattac cgcgagccgt tcctgaatcc
     2521 tgttgaccgc gagccactgt ggcgcttccc aaacgagctg ccaatcgccg gtgagccagc
     2581 gaacatcgtc gcgctggtcg aagaatacat ggactggctg caccagtccc ctgtcccgaa
     2641 gctgctgttc tggggcaccc caggcgttct gatcccaccg gccgaagccg ctcgcctggc
     2701 caaaagcctg cctaactgca aggctgtgga catcggcccg ggtctgaatc tgctgcaaga
     2761 agacaacccg gacctgatcg gcagcgagat cgcgcgctgg ctgtctactc tggagatttc
     2821 cggtcatcat caccatcacc actaatagtc aaccgctgag caataactag cataacccct
     2881 tggggcctct aaacgggtct tgaggggttt tttgctgaaa ggaggaacta tatccggttc
     2941 gcttgctgtc cataaaaccg cccagtctag ctatcgccat gtaagcccac tgcaagctac
     3001 ctgctttctc tttgcgcttg cgttttccct tgtccagata gcccagtagc tgacattcat
     3061 ccggggtcag caccgtttct gcggactggc tttctacgta atggtttctt agacgtcagg
     3121 tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc
     3181 aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag
     3241 gaagagtatg agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg
     3301 ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta aaagatgctg aagatcagtt
     3361 gggtgcacga gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt
     3421 tcgccccgaa gaacgttttc caatgatgag cactttcaaa gttctgctat gtggcgcggt
     3481 attatcccgt attgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa
     3541 tgacttggtt gagtactcac cagtcacaga aaagcatctt acggatggca tgacagtaag
     3601 agaattatgc agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac
     3661 aactatcgga ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac
     3721 tcgccttgat cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac
     3781 cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg aactacttac
     3841 tctagcttcc cggcaacaat taatagactg gatggaggcg gataaagttg caggaccact
     3901 tctgcgctcg gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg
     3961 tgggtctcgc ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt
     4021 tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat
     4081 aggtgcctca ctgattaagc attggtaatt cgaaatgacc gaccaagcga cgcccaaccg
     4141 gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga
     4201 aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg
     4261 gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag
     4321 aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc
     4381 gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg
     4441 ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt
     4501 cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc
     4561 ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc
     4621 actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg
     4681 tggcctaact acggctacac tagaaggaca gtatttggta tctgcgctct gctgaagcca
     4741 gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc
     4801 ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatt tcaagaagat
     4861 cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggcat
     4921 atgatggtag tgtggggtct ccccatgcga gagtagggaa ctgccaggca tcaaataaaa
     4981 cgaaaggctc agtcgaaaga ctgggccttt cgttttatct gttgtttgtc ggtgaacgct
     5041 ctcctgagta ggacaaatcc gccgggagcg gatttgaacg ttgcgaagca acggcccgga
     5101 gggtgg
//
