LOCUS       sequence-345490-        6093 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 6093)
  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 6093)
  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_IRP1-Halo
FEATURES             Location/Qualifiers
     source          1..6093
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        94..198
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(199..267,268..1059)
                     /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(417..436)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      complement(1217..1805)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1297..1316)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      1882..1968
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      2060..2087
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     primer_bind     2169..2188
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2169..2187
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      2245..2254
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             4948..4968
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="EDLYFQS"
     CDS             4984..5868
                     /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             5869..5886
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(5897..5915)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      5911..5958
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
ORIGIN
        1 ctgacattca tccggggtca gcaccgtttc tgcggactgg ctttctacgt aatggtttct
       61 tagacgtcag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc
      121 taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa
      181 tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat tccctttttt
      241 gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct
      301 gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag cggtaagatc
      361 cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcactttcaa agttctgcta
      421 tgtggcgcgg tattatcccg tattgacgcc gggcaagagc aactcggtcg ccgcatacac
      481 tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct tacggatggc
      541 atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac
      601 ttacttctga caactatcgg aggaccgaag gagctaaccg cttttttgca caacatgggg
      661 gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat accaaacgac
      721 gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact attaactggc
      781 gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc ggataaagtt
      841 gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga taaatctgga
      901 gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg taagccctcc
      961 cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag
     1021 atcgctgaga taggtgcctc actgattaag cattggtaat tcgaaatgac cgaccaagcg
     1081 acgcccaacc ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg
     1141 ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg
     1201 ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac
     1261 gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg
     1321 gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct
     1381 ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg
     1441 tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct
     1501 gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac
     1561 tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt
     1621 tcttgaagtg gtggcctaac tacggctaca ctagaaggac agtatttggt atctgcgctc
     1681 tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca
     1741 ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat
     1801 ttcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac
     1861 gttaagggca tagccaggca tcaaataaaa cgaaaggctc agtcgaaaga ctgggccttt
     1921 cgttttatct gttgtttgtc ggtgaacgct ctcctgagta ggacaaatcc gccgggagcg
     1981 gatttgaacg ttgcgaagca acggcccgga gggtggcggg caggacgccc gccataaact
     2041 gccaggcatc aaattaagca gaaggccatc ctgacggatg gcctttttgc gtttctacaa
     2101 actcttttgt ttatttttct aaatacattc aaatatgtat ccgctcatgg atctcgatcc
     2161 cgcgaaatta atacgactca ctatagggta ttttaacttt actaaggagc gatcgcttcc
     2221 gaattcctac cgcggatatc tagagccacc atggcgagca acccattcgc acaccttgct
     2281 gagccattgg atcctgtaca accaggaaag aaattcttca atttgaataa attggaggat
     2341 tcaagatatg ggcgcttacc attttcgatc agagttcttc tggaagcagc cattcggaat
     2401 tgtgatgagt ttttggtgaa gaaacaggat attgaaaata ttctacattg gaatgtcacg
     2461 cagcacaaga acatagaagt gccatttaag cctgctcgtg tcatcctgca ggactttacg
     2521 ggtgtgcccg ctgtggttga ctttgctgca atgcgtgatg ctgtgaaaaa gttaggagga
     2581 gatccagaga aaataaaccc tgtctgccct gctgatcttg taatagatca ttccatccag
     2641 gttgatttca acagaagggc agacagttta cagaagaatc aagacctgga atttgaaaga
     2701 aatagagagc gatttgaatt tttaaagtgg ggttcccagg cttttcacaa catgcggatt
     2761 attccccctg gctcaggaat catccaccag gtgaatttgg aatatttggc aagagtggta
     2821 tttgatcagg atggatatta ttacccagac agcctcgtgg gcacagactc gcacactacc
     2881 atgattgatg gcttgggcat tcttggttgg ggtgtcggtg gtattgaagc agaagctgtc
     2941 atgctgggtc agccaatcag tatggtgctt cctcaggtga ttggctacag gctgatgggg
     3001 aagccccacc ctctggtaac atccactgac atcgtgctca ccattaccaa gcacctccgc
     3061 caggttgggg tagtgggcaa atttgtcgag ttcttcgggc ctggagtagc ccagttgtcc
     3121 attgctgacc gagctacgat tgctaacatg tgtccagagt acggagcaac tgctgccttt
     3181 ttcccagttg atgaagttag tatcacgtac ctggtgcaaa caggtcgtga tgaagaaaaa
     3241 ttaaagtata ttaaaaaata tcttcaggct gtaggaatgt ttcgagattt caatgaccct
     3301 tctcaagacc cagacttcac ccaggttgtg gaattagatt tgaaaacagt agtgccttgc
     3361 tgtagtggac ccaaaaggcc tcaggacaaa gttgctgtgt ccgacatgaa aaaggacttt
     3421 gagagctgcc ttggagccaa gcaaggattt aaaggattcc aagttgctcc tgaacatcat
     3481 aatgaccata agacctttat ctatgataac actgagttca cccttgctca tggttctgtg
     3541 gtcattgctg ccattactag ctgcacaaac accagtaatc cgtctgtgat gttaggggca
     3601 ggattgttag caaagaaagc tgtggatgct ggcctgaacg tgatgcctta catcaaaact
     3661 agcctgtctc ctgggagtgg cgtggtcacc tactacctac aagaaagcgg agtcatgcct
     3721 tatctgtctc agcttgggtt tgacgtggtg ggctatggct gcatgacctg cattggcaac
     3781 agtgggcctt tacctgaacc tgtggtagaa gccatcacac agggagacct tgtagctgtt
     3841 ggagtactat ctggaaacag gaattttgaa ggtcgagttc accccaacac ccgggccaac
     3901 tatttagcct ctcccccctt agtaatagca tatgcaattg ctggaaccat cagaatcgac
     3961 tttgagaaag agccattggg agtaaatgca aagggacagc aggtatttct gaaagatatc
     4021 tggccgacta gagacgagat ccaggcagtg gagcgtcagt atgtcatccc ggggatgttt
     4081 aaggaagtct atcagaaaat agagactgtg aatgaaagct ggaatgcctt agcaacccca
     4141 tcagataagc tgtttttctg gaattccaaa tctacgtata tcaaatcacc accattcttt
     4201 gaaaacctga ctttggatct tcagccccct aaatctatag tggatgccta tgtgctgcta
     4261 aatttgggag attcggtaac aactgaccac atctccccag ctggaaatat tgcaagaaac
     4321 agtcctgctg ctcgctactt aactaacaga ggcctaactc cacgagaatt caactcctat
     4381 ggctcccgcc gaggtaatga cgccgtcatg gcacggggaa catttgccaa cattcgcttg
     4441 ttaaacagat ttttgaacaa gcaggcacca cagactatcc atctgccttc tggggaaatc
     4501 cttgatgtgt ttgatgctgc tgagcggtac cagcaggcag gccttcccct gatcgttctg
     4561 gctggcaaag agtacggtgc aggcagctcc cgagactggg cagctaaggg ccctttcctg
     4621 ctgggaatca aagccgtcct ggccgagagc tacgagcgca ttcaccgcag taacctggtt
     4681 gggatgggtg tgatcccact tgaatatctc cctggtgaga atgcagatgc cctggggctc
     4741 acagggcaag aacgatacac tatcattatt ccagaaaacc tcaaaccaca aatgaaagtc
     4801 caggtcaagc tggatactgg caagaccttc caggctgtca tgaggtttga cactgatgtg
     4861 gagctcactt atttcctcaa cgggggcatc ctcaactaca tgatccgcaa gatggccaag
     4921 ggcggctccg gcctcgagcc aaccactgag gatctgtact ttcagagcga taacgatgga
     4981 tccgaaatcg gtactggctt tccattcgac ccccattatg tggaagtcct gggcgagcgc
     5041 atgcactacg tcgatgttgg tccgcgcgat ggcacccctg tgctgttcct gcacggtaac
     5101 ccgacctcct cctacgtgtg gcgcaacatc atcccgcatg ttgcaccgac ccatcgctgc
     5161 attgctccag acctgatcgg tatgggcaaa tccgacaaac cagacctggg ttatttcttc
     5221 gacgaccacg tccgcttcat ggatgccttc atcgaagccc tgggtctgga agaggtcgtc
     5281 ctggtcattc acgactgggg ctccgctctg ggtttccact gggccaagcg caatccagag
     5341 cgcgtcaaag gtattgcatt tatggagttc atccgcccta tcccgacctg ggacgaatgg
     5401 ccagaatttg cccgcgagac cttccaggcc ttccgcacca ccgacgtcgg ccgcaagctg
     5461 atcatcgatc agaacgtttt tatcgagggt acgctgccga tgggtgtcgt ccgcccgctg
     5521 actgaagtcg agatggacca ttaccgcgag ccgttcctga atcctgttga ccgcgagcca
     5581 ctgtggcgct tcccaaacga gctgccaatc gccggtgagc cagcgaacat cgtcgcgctg
     5641 gtcgaagaat acatggactg gctgcaccag tcccctgtcc cgaagctgct gttctggggc
     5701 accccaggcg ttctgatccc accggccgaa gccgctcgcc tggccaaaag cctgcctaac
     5761 tgcaaggctg tggacatcgg cccgggtctg aatctgctgc aagaagacaa cccggacctg
     5821 atcggcagcg agatcgcgcg ctggctgtct actctggaga tttccggtca tcatcaccat
     5881 caccactaat agtcaaccgc tgagcaataa ctagcataac cccttggggc ctctaaacgg
     5941 gtcttgaggg gttttttgct gaaaggagga actatatccg gttcgcttgc tgtccataaa
     6001 accgcccagt ctagctatcg ccatgtaagc ccactgcaag ctacctgctt tctctttgcg
     6061 cttgcgtttt cccttgtcca gatagcccag tag
//
