LOCUS       sequence-345583-        4992 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 4992)
  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 4992)
  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_FAM98A-Halo
FEATURES             Location/Qualifiers
     source          1..4992
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      37..64
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     primer_bind     146..165
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        146..164
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      1317..1326
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1783..1803
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="EDLYFQS"
     CDS             1819..2703
                     /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             2704..2721
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(2732..2750)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      2746..2793
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        3022..3126
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3127..3195,3196..3987)
                     /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(3345..3364)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      complement(4145..4733)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4225..4244)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      4851..4937
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
ORIGIN
        1 gacgcccgcc ataaactgcc aggcatcaaa ttaagcagaa ggccatcctg acggatggcc
       61 tttttgcgtt tctacaaact cttttgttta tttttctaaa tacattcaaa tatgtatccg
      121 ctcatggatc tcgatcccgc gaaattaata cgactcacta tagggtattt taactttact
      181 aaggagcgat cgcttccgaa ttcctaccgc ggatatatgg agtgtgacct catggagact
      241 gacatcttgg agtcgttgga agatctaggt tacaagggcc cattgttgga agatggagcg
      301 ctctctcagg cagtctctgc tggagccagt tcccccgagt ttaccaaact ctgtgcttgg
      361 ctggtgtctg aattaagagt gctctgtaaa ctagaggaaa acgtgcaagc aactaacagt
      421 ccgagtgaag ctgaagaatt ccagcttgag gtgagtgggc tactagggga gatgaactgc
      481 ccgtatcttt cactgacatc tggggatgtg accaagcgcc ttctcattca gaagaactgc
      541 ctcctcttgc tcacatacct catctcagaa ctagaagctg ccagaatgct ctgtgtgaat
      601 gctcctccaa aaaaagctca agaaggaggc ggtagtgagg tctttcaaga gttgaaaggc
      661 atatgtattg ctctaggaat gtccaaacct ccagccaata taactatgtt ccaattcttc
      721 agcgggattg aaaaaaaatt aaaggaaaca ttagcaaaag ttccacctaa tcatgtggga
      781 aagcctttac tgaagaagcc aatgggacca gcccactggg aaaagataga agcaattaac
      841 caagccatag ccaatgaata tgaagtccgg agaaagctgc taataaaacg tttggatgtc
      901 actgtacaat cctttggctg gtctgacaga gctaagagcc agacagaaaa attagccaag
      961 gtttaccagc cgaaacgttc agtcttatcc cctaaaacta ctatttctgt tgcccatctt
     1021 ttggctgcaa ggcaggactt gtcaaagatt ttaaggacaa gcagcggctc tataagagaa
     1081 aagactgcct gtgccatcaa taaggtgttg atgggcaggg tgcctgacag aggtggtaga
     1141 cccaatgaaa tcgaacctcc acccccagag atgccaccgt ggcagaagag gcaagatggc
     1201 ccccagcagc aaacaggagg ccgaggagga gggagaggtg gctatgaaca ttcctcatac
     1261 ggaggacgag gaggtcatga acaaggaggc gggagaggtg gacgtggtgg ctatgaccat
     1321 ggtggccgag ggggaggaag aggaaataag catcaaggag gctggacaga tggagggagt
     1381 ggtggaggag gtggctacca agatggtggt tatcgagatt caggtttcca gccaggtggc
     1441 tatcatggtg gccacagcag tggtggctat caaggcggag gttatggtgg cttccaaaca
     1501 tcttcttcat atacaggaag tggataccag ggtggtggct accagcagga caatagatac
     1561 caagatggcg ggcaccatgg tgatcgtggt ggtggtcgtg gtgggcgagg tggtcgtgga
     1621 ggccgaggtg gtcgtgcagg ccagggagga ggctggggag gaagagggag ccagaattat
     1681 caccaagggg gtcaatttga acagcatttc cagcatggag gttatcagta taatcattct
     1741 ggatttggac agggaagaca ttacactagt gggccaacca ctgaggatct gtactttcag
     1801 agcgataacg atggatccga aatcggtact ggctttccat tcgaccccca ttatgtggaa
     1861 gtcctgggcg agcgcatgca ctacgtcgat gttggtccgc gcgatggcac ccctgtgctg
     1921 ttcctgcacg gtaacccgac ctcctcctac gtgtggcgca acatcatccc gcatgttgca
     1981 ccgacccatc gctgcattgc tccagacctg atcggtatgg gcaaatccga caaaccagac
     2041 ctgggttatt tcttcgacga ccacgtccgc ttcatggatg ccttcatcga agccctgggt
     2101 ctggaagagg tcgtcctggt cattcacgac tggggctccg ctctgggttt ccactgggcc
     2161 aagcgcaatc cagagcgcgt caaaggtatt gcatttatgg agttcatccg ccctatcccg
     2221 acctgggacg aatggccaga atttgcccgc gagaccttcc aggccttccg caccaccgac
     2281 gtcggccgca agctgatcat cgatcagaac gtttttatcg agggtacgct gccgatgggt
     2341 gtcgtccgcc cgctgactga agtcgagatg gaccattacc gcgagccgtt cctgaatcct
     2401 gttgaccgcg agccactgtg gcgcttccca aacgagctgc caatcgccgg tgagccagcg
     2461 aacatcgtcg cgctggtcga agaatacatg gactggctgc accagtcccc tgtcccgaag
     2521 ctgctgttct ggggcacccc aggcgttctg atcccaccgg ccgaagccgc tcgcctggcc
     2581 aaaagcctgc ctaactgcaa ggctgtggac atcggcccgg gtctgaatct gctgcaagaa
     2641 gacaacccgg acctgatcgg cagcgagatc gcgcgctggc tgtctactct ggagatttcc
     2701 ggtcatcatc accatcacca ctaatagtca accgctgagc aataactagc ataacccctt
     2761 ggggcctcta aacgggtctt gaggggtttt ttgctgaaag gaggaactat atccggttcg
     2821 cttgctgtcc ataaaaccgc ccagtctagc tatcgccatg taagcccact gcaagctacc
     2881 tgctttctct ttgcgcttgc gttttccctt gtccagatag cccagtagct gacattcatc
     2941 cggggtcagc accgtttctg cggactggct ttctacgtaa tggtttctta gacgtcaggt
     3001 ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca
     3061 aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg
     3121 aagagtatga gtattcaaca tttccgtgtc gcccttattc ccttttttgc ggcattttgc
     3181 cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga agatcagttg
     3241 ggtgcacgag tgggttacat cgaactggat ctcaacagcg gtaagatcct tgagagtttt
     3301 cgccccgaag aacgttttcc aatgatgagc actttcaaag ttctgctatg tggcgcggta
     3361 ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat
     3421 gacttggttg agtactcacc agtcacagaa aagcatctta cggatggcat gacagtaaga
     3481 gaattatgca gtgctgccat aaccatgagt gataacactg cggccaactt acttctgaca
     3541 actatcggag gaccgaagga gctaaccgct tttttgcaca acatggggga tcatgtaact
     3601 cgccttgatc gttgggaacc ggagctgaat gaagccatac caaacgacga gcgtgacacc
     3661 acgatgcctg tagcaatggc aacaacgttg cgcaaactat taactggcga actacttact
     3721 ctagcttccc ggcaacaatt aatagactgg atggaggcgg ataaagttgc aggaccactt
     3781 ctgcgctcgg cccttccggc tggctggttt attgctgata aatctggagc cggtgagcgt
     3841 gggtctcgcg gtatcattgc agcactgggg ccagatggta agccctcccg tatcgtagtt
     3901 atctacacga cggggagtca ggcaactatg gatgaacgaa atagacagat cgctgagata
     3961 ggtgcctcac tgattaagca ttggtaattc gaaatgaccg accaagcgac gcccaaccgg
     4021 tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa
     4081 agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg
     4141 cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga
     4201 ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg
     4261 tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg
     4321 gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc
     4381 gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg
     4441 gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca
     4501 ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt
     4561 ggcctaacta cggctacact agaaggacag tatttggtat ctgcgctctg ctgaagccag
     4621 ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg
     4681 gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggattt caagaagatc
     4741 ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggcata
     4801 tgatggtagt gtggggtctc cccatgcgag agtagggaac tgccaggcat caaataaaac
     4861 gaaaggctca gtcgaaagac tgggcctttc gttttatctg ttgtttgtcg gtgaacgctc
     4921 tcctgagtag gacaaatccg ccgggagcgg atttgaacgt tgcgaagcaa cggcccggag
     4981 ggtggcgggc ag
//
