LOCUS       sequence-344841-        4497 bp    DNA     circular SYN 14-APR-2025
DEFINITION  Production of HaloTag fusion protein.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4497)
  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 4497)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 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_PRMT1-Halo
FEATURES             Location/Qualifiers
     source          1..4497
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      55..141
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      233..260
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     primer_bind     342..361
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        342..360
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             1520..2404
                     /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             2405..2422
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(2433..2451)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      2447..2494
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        2723..2827
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2828..2896,2897..3688)
                     /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(3046..3065)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      complement(3846..4434)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3926..3945)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 catatgatgg tagtgtgggg tctccccatg cgagagtagg gaactgccag gcatcaaata
       61 aaacgaaagg ctcagtcgaa agactgggcc tttcgtttta tctgttgttt gtcggtgaac
      121 gctctcctga gtaggacaaa tccgccggga gcggatttga acgttgcgaa gcaacggccc
      181 ggagggtggc gggcaggacg cccgccataa actgccaggc atcaaattaa gcagaaggcc
      241 atcctgacgg atggcctttt tgcgtttcta caaactcttt tgtttatttt tctaaataca
      301 ttcaaatatg tatccgctca tggatctcga tcccgcgaaa ttaatacgac tcactatagg
      361 gtattttaac tttactaagg agcgatcgct tccgaattcc taccgcggat atatggcggc
      421 agccgaggcc gcgaactgca tcatggaggt gtcctgtggc caggcggaaa gcagtgagaa
      481 gcccaacgct gaggacatga catccaaaga ttactacttt gactcctacg cacactttgg
      541 catccacgag gagatgctga aggacgaggt gcgcaccctc acttaccgca actccatgtt
      601 tcataaccgg cacctcttca aggacaaggt ggtgctggac gtcggctcgg gcaccggcat
      661 cctctgcatg tttgctgcca aggccggggc ccgcaaggtc atcgggatcg agtgttccag
      721 tatctctgat tatgcggtga agatcgtcaa agccaacaag ttagaccacg tggtgaccat
      781 catcaagggg aaggtggagg aggtggagct cccagtggag aaggtggaca tcatcatcag
      841 cgagtggatg ggctactgcc tcttctacga gtccatgctc aacaccgtgc tctatgcccg
      901 ggacaagtgg ctggcgcccg atggcctcat cttcccagac cgggccacgc tgtatgtgac
      961 ggccatcgag gaccggcagt acaaagacta caagatccac tggtgggaga acgtgtatgg
     1021 cttcgacatg tcttgcatca aagatgtggc cattaaggag cccctagtgg atgtcgtgga
     1081 ccccaaacag ctggtcacca acgcctgcct cataaaggag gtggacatct ataccgtcaa
     1141 ggtggaagac ctgaccttca cctccccgtt ctgcctgcaa gtgaagcgga atgactacgt
     1201 gcacgccctg gtggcctact tcaacatcga gttcacacgc tgccacaaga ggaccggctt
     1261 ctccaccagc cccgagtccc cgtacacgca ctggaagcag acggtgttct acatggagga
     1321 ctacctgacc gtgaagacgg gcgaggagat cttcggcacc atcggcatgc ggcccaacgc
     1381 caagaacaac cgggacctgg acttcaccat cgacctggac ttcaagggcc agctgtgcga
     1441 gctgtcctgc tccaccgact accggatgcg cgggccaacc actgaggatc tgtcctttca
     1501 gagcgataac gatggatccg aaatcggtac tggctttcca ttcgaccccc attatgtgga
     1561 agtcctgggc gagcgcatgc actacgtcga tgttggtccg cgcgatggca cccctgtgct
     1621 gttcctgcac ggtaacccga cctcctccta cgtgtggcgc aacatcatcc cgcatgttgc
     1681 accgacccat cgctgcattg ctccagacct gatcggtatg ggcaaatccg acaaaccaga
     1741 cctgggttat ttcttcgacg accacgtccg cttcatggat gccttcatcg aagccctggg
     1801 tctggaagag gtcgtcctgg tcattcacga ctggggctcc gctctgggtt tccactgggc
     1861 caagcgcaat ccagagcgcg tcaaaggtat tgcatttatg gagttcatcc gccctatccc
     1921 gacctgggac gaatggccag aatttgcccg cgagaccttc caggccttcc gcaccaccga
     1981 cgtcggccgc aagctgatca tcgatcagaa cgtttttatc gagggtacgc tgccgatggg
     2041 tgtcgtccgc ccgctgactg aagtcgagat ggaccattac cgcgagccgt tcctgaatcc
     2101 tgttgaccgc gagccactgt ggcgcttccc aaacgagctg ccaatcgccg gtgagccagc
     2161 gaacatcgtc gcgctggtcg aagaatacat ggactggctg caccagtccc ctgtcccgaa
     2221 gctgctgttc tggggcaccc caggcgttct gatcccaccg gccgaagccg ctcgcctggc
     2281 caaaagcctg cctaactgca aggctgtgga catcggcccg ggtctgaatc tgctgcaaga
     2341 agacaacccg gacctgatcg gcagcgagat cgcgcgctgg ctgtctactc tggagatttc
     2401 cggtcatcat caccatcacc actaatagtc aaccgctgag caataactag cataacccct
     2461 tggggcctct aaacgggtct tgaggggttt tttgctgaaa ggaggaacta tatccggttc
     2521 gcttgctgtc cataaaaccg cccagtctag ctatcgccat gtaagcccac tgcaagctac
     2581 ctgctttctc tttgcgcttg cgttttccct tgtccagata gcccagtagc tgacattcat
     2641 ccggggtcag caccgtttct gcggactggc tttctacgta atggtttctt agacgtcagg
     2701 tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc
     2761 aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag
     2821 gaagagtatg agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg
     2881 ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta aaagatgctg aagatcagtt
     2941 gggtgcacga gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt
     3001 tcgccccgaa gaacgttttc caatgatgag cactttcaaa gttctgctat gtggcgcggt
     3061 attatcccgt attgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa
     3121 tgacttggtt gagtactcac cagtcacaga aaagcatctt acggatggca tgacagtaag
     3181 agaattatgc agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac
     3241 aactatcgga ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac
     3301 tcgccttgat cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac
     3361 cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg aactacttac
     3421 tctagcttcc cggcaacaat taatagactg gatggaggcg gataaagttg caggaccact
     3481 tctgcgctcg gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg
     3541 tgggtctcgc ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt
     3601 tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat
     3661 aggtgcctca ctgattaagc attggtaatt cgaaatgacc gaccaagcga cgcccaaccg
     3721 gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga
     3781 aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg
     3841 gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag
     3901 aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc
     3961 gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg
     4021 ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt
     4081 cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc
     4141 ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc
     4201 actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg
     4261 tggcctaact acggctacac tagaaggaca gtatttggta tctgcgctct gctgaagcca
     4321 gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc
     4381 ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatt tcaagaagat
     4441 cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaaggg
//
