LOCUS       sequence-345487-        3763 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 3763)
  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 3763)
  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: pH6HTN_Halo-STMN1
FEATURES             Location/Qualifiers
     source          1..3763
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      34..61
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     primer_bind     143..162
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        143..161
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             198..215
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             216..1103
                     /codon_start=1
                     /product="modified bacterial dehalogenase that forms 
                     covalent bonds with chloroalkane derivatives"
                     /label=HaloTag(R)
                     /translation="AEIGTGFPFDPHYVEVLGERMHYVDVGPRDGTPVLFLHGNPTSSY
                     VWRNIIPHVAPTHRCIAPDLIGMGKSDKPDLGYFFDDHVRFMDAFIEALGLEEVVLVIH
                     DWGSALGFHWAKRNPERVKGIAFMEFIRPIPTWDEWPEFARETFQAFRTTDVGRKLIID
                     QNVFIEGTLPMGVVRPLTEVEMDHYREPFLNPVDREPLWRFPNELPIAGEPANIVALVE
                     EYMDWLHQSPVPKLLFWGTPGVLIPPAEAARLAKSLPNCKAVDIGPGLNLLQEDNPDLI
                     GSEIARWLSTLEISG"
     CDS             1116..1136
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="EDLYFQS"
     primer_bind     complement(1674..1692)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1688..1735
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        1964..2068
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2069..2137,2138..2929)
                     /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(2287..2306)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      complement(3087..3675)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3167..3186)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gcccgccata aactgccagg catcaaatta agcagaaggc catcctgacg gatggccttt
       61 ttgcgtttct acaaactctt ttgtttattt ttctaaatac attcaaatat gtatccgctc
      121 atggatctcg atcccgcgaa attaatacga ctcactatag ggtattttaa ctttactaag
      181 gagaattcat catgaaacat catcaccatc accacgcaga aatcggtact ggctttccat
      241 tcgaccccca ttatgtggaa gtcctgggcg agcgcatgca ctacgtcgat gttggtccgc
      301 gcgatggcac ccctgtgctg ttcctgcacg gtaacccgac ctcctcctac gtgtggcgca
      361 acatcatccc gcatgttgca ccgacccatc gctgcattgc tccagacctg atcggtatgg
      421 gcaaatccga caaaccagac ctgggttatt tcttcgacga ccacgtccgc ttcatggatg
      481 ccttcatcga agccctgggt ctggaagagg tcgtcctggt cattcacgac tggggctccg
      541 ctctgggttt ccactgggcc aagcgcaatc cagagcgcgt caaaggtatt gcatttatgg
      601 agttcatccg ccctatcccg acctgggacg aatggccaga atttgcccgc gagaccttcc
      661 aggccttccg caccaccgac gtcggccgca agctgatcat cgatcagaac gtttttatcg
      721 agggtacgct gccgatgggt gtcgtccgcc cgctgactga agtcgagatg gaccattacc
      781 gcgagccgtt cctgaatcct gttgaccgcg agccactgtg gcgcttccca aacgagctgc
      841 caatcgccgg tgagccagcg aacatcgtcg cgctggtcga agaatacatg gactggctgc
      901 accagtcccc tgtcccgaag ctgctgttct ggggcacccc aggcgttctg atcccaccgg
      961 ccgaagccgc tcgcctggcc aaaagcctgc ctaactgcaa ggctgtggac atcggcccgg
     1021 gtctgaatct gctgcaagaa gacaacccgg acctgatcgg cagcgagatc gcgcgctggc
     1081 tgtcgacgct cgagatttcc ggcgagccaa ccactgagga tctgtacttt cagagcgata
     1141 acgcgatcgc ttccgaattc agagctcaac cgcggatatc tagagcctct tctgatatcc
     1201 aggtgaaaga actggagaag cgtgcctcag gccaggcttt tgagctgatt ctcagccctc
     1261 ggtcaaaaga atctgttcca gaattccccc tttcccctcc aaagaagaag gatctttccc
     1321 tggaggaaat tcagaagaaa ttagaagctg cagaagaaag acgcaagtcc catgaagctg
     1381 aggtcttgaa gcagctggct gagaaacgag agcacgagaa agaagtgctt cagaaggcaa
     1441 tagaagagaa caacaacttc agtaaaatgg cagaagagaa actgacccac aaaatggaag
     1501 ctaataaaga gaaccgagag gcacaaatgg ctgccaaact ggaacgtttg cgagagaagg
     1561 ataagcacat tgaagaagtg cggaagaaca aagaatccaa agaccctgct gacgagactg
     1621 aagctgacta acctgcagga ttttgcggcc gcttgctgag ttggctgctg ccaccgctga
     1681 gcaataacta gcataacccc ttggggcctc taaacgggtc ttgaggggtt ttttgctgaa
     1741 aggaggaact atatccggtt cgcttgctgt ccataaaacc gcccagtcta gctatcgcca
     1801 tgtaagccca ctgcaagcta cctgctttct ctttgcgctt gcgttttccc ttgtccagat
     1861 agcccagtag ctgacattca tccggggtca gcaccgtttc tgcggactgg ctttctacgt
     1921 aatggtttct tagacgtcag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg
     1981 tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat
     2041 gcttcaataa tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat
     2101 tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt
     2161 aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag
     2221 cggtaagatc cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcactttcaa
     2281 agttctgcta tgtggcgcgg tattatcccg tattgacgcc gggcaagagc aactcggtcg
     2341 ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct
     2401 tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac
     2461 tgcggccaac ttacttctga caactatcgg aggaccgaag gagctaaccg cttttttgca
     2521 caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat
     2581 accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact
     2641 attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc
     2701 ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga
     2761 taaatctgga gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg
     2821 taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg
     2881 aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaat tcgaaatgac
     2941 cgaccaagcg acgcccaacc ggtatcagct cactcaaagg cggtaatacg gttatccaca
     3001 gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac
     3061 cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac
     3121 aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg
     3181 tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac
     3241 ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat
     3301 ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag
     3361 cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac
     3421 ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt
     3481 gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaaggac agtatttggt
     3541 atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc
     3601 aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga
     3661 aaaaaaggat ttcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac
     3721 gaaaactcac gttaagggca tatgtttggg tggcgggcag gac
//
