LOCUS       sequence-344842-        4398 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 4398)
  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 4398)
  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_HNRNPA1-Halo
FEATURES             Location/Qualifiers
     source          1..4398
                     /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             1201..1212
                     /codon_start=1
                     /product="WELQut protease recognition and cleavage site"
                     /label=WELQut site
                     /translation="WELQ"
     CDS             1385..1405
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="EDLYFQS"
     CDS             1421..2305
                     /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             2306..2323
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(2334..2352)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      2348..2395
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        2624..2728
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2729..2797,2798..3589)
                     /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(2947..2966)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      complement(3747..4335)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3827..3846)
                     /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 atatgtctaa
      421 gtcagagtct cctaaagagc ccgaacagct gaggaagctc ttcattggag ggttgagctt
      481 tgaaacaact gatgagagcc tgaggagcca ttttgagcaa tggggaacgc tcacggactg
      541 tgtggtaatg agagatccaa acaccaagcg ctccaggggc tttgggtttg tcacatatgc
      601 cactgtggag gaggtggatg cagctatgaa tgcaaggcca cacaaggtgg atggaagagt
      661 tgtggaacca aagagagctg tctccagaga agattctcaa agaccaggtg cccacttaac
      721 tgtgaaaaag atatttgttg gtggcattaa agaagacact gaagaacatc acctaagaga
      781 ttattttgaa cagtatggaa aaattgaagt gattgaaatc atgactgacc gaggcagtgg
      841 caagaaaagg ggctttgcct ttgtaacctt tgacgaccat gactccgtgg ataagattgt
      901 cattcagaaa taccatactg tgaatggcca caactgtgaa gttagaaaag ccctgtcaaa
      961 gcaagagatg gctagtgctt catccagcca aagaggtcga agtggttctg gaaactttgg
     1021 tggtggtcgt ggaggtggtt tcggtgggaa tgacaacttc ggtcgtggag gaaacttcag
     1081 tggtcgtggt ggctttggtg gcagccgtgg tggtggtgga tatggtggca gtggggatgg
     1141 ctataatgga tttggtaatg atggaagcaa ttttggaggt ggtggaagct acaatgattt
     1201 tgggaattac aacaatcagt cttcaaattt tggacccatg aagggaggaa attttggagg
     1261 cagaagctct ggcccctatg gcggtggagg ccaatacttt gcaaaaccac gaaaccaagg
     1321 tggctatggc ggttccagca gcagcagtag ctatggcagt ggcagaagat ttgggccaac
     1381 cactgaggat ctgtactttc agagcgataa cgatggatcc gaaatcggta ctggctttcc
     1441 attcgacccc cattatgtgg aagtcctggg cgagcgcatg cactacgtcg atgttggtcc
     1501 gcgcgatggc acccctgtgc tgttcctgca cggtaacccg acctcctcct acgtgtggcg
     1561 caacatcatc ccgcatgttg caccgaccca tcgctgcatt gctccagacc tgatcggtat
     1621 gggcaaatcc gacaaaccag acctgggtta tttcttcgac gaccacgtcc gcttcatgga
     1681 tgccttcatc gaagccctgg gtctggaaga ggtcgtcctg gtcattcacg actggggctc
     1741 cgctctgggt ttccactggg ccaagcgcaa tccagagcgc gtcaaaggta ttgcatttat
     1801 ggagttcatc cgccctatcc cgacctggga cgaatggcca gaatttgccc gcgagacctt
     1861 ccaggccttc cgcaccaccg acgtcggccg caagctgatc atcgatcaga acgtttttat
     1921 cgagggtacg ctgccgatgg gtgtcgtccg cccgctgact gaagtcgaga tggaccatta
     1981 ccgcgagccg ttcctgaatc ctgttgaccg cgagccactg tggcgcttcc caaacgagct
     2041 gccaatcgcc ggtgagccag cgaacatcgt cgcgctggtc gaagaataca tggactggct
     2101 gcaccagtcc cctgtcccga agctgctgtt ctggggcacc ccaggcgttc tgatcccacc
     2161 ggccgaagcc gctcgcctgg ccaaaagcct gcctaactgc aaggctgtgg acatcggccc
     2221 gggtctgaat ctgctgcaag aagacaaccc ggacctgatc ggcagcgaga tcgcgcgctg
     2281 gctgtctact ctggagattt ccggtcatca tcaccatcac cactaatagt caaccgctga
     2341 gcaataacta gcataacccc ttggggcctc taaacgggtc ttgaggggtt ttttgctgaa
     2401 aggaggaact atatccggtt cgcttgctgt ccataaaacc gcccagtcta gctatcgcca
     2461 tgtaagccca ctgcaagcta cctgctttct ctttgcgctt gcgttttccc ttgtccagat
     2521 agcccagtag ctgacattca tccggggtca gcaccgtttc tgcggactgg ctttctacgt
     2581 aatggtttct tagacgtcag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg
     2641 tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat
     2701 gcttcaataa tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat
     2761 tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt
     2821 aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag
     2881 cggtaagatc cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcactttcaa
     2941 agttctgcta tgtggcgcgg tattatcccg tattgacgcc gggcaagagc aactcggtcg
     3001 ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct
     3061 tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac
     3121 tgcggccaac ttacttctga caactatcgg aggaccgaag gagctaaccg cttttttgca
     3181 caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat
     3241 accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact
     3301 attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc
     3361 ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga
     3421 taaatctgga gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg
     3481 taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg
     3541 aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaat tcgaaatgac
     3601 cgaccaagcg acgcccaacc ggtatcagct cactcaaagg cggtaatacg gttatccaca
     3661 gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac
     3721 cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac
     3781 aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg
     3841 tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac
     3901 ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat
     3961 ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag
     4021 cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac
     4081 ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt
     4141 gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaaggac agtatttggt
     4201 atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc
     4261 aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga
     4321 aaaaaaggat ttcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac
     4381 gaaaactcac gttaaggg
//
