LOCUS       sequence-345580-        5400 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 5400)
  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 5400)
  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_HSPA5-Halo
FEATURES             Location/Qualifiers
     source          1..5400
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      29..56
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     primer_bind     138..157
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        138..156
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             2183..2203
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="EDLYFQS"
     CDS             2219..3103
                     /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             3104..3121
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(3132..3150)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      3146..3193
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        3422..3526
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3527..3595,3596..4387)
                     /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(3745..3764)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      complement(4545..5133)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4625..4644)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      5251..5337
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
ORIGIN
        1 ccataaactg ccaggcatca aattaagcag aaggccatcc tgacggatgg cctttttgcg
       61 tttctacaaa ctcttttgtt tatttttcta aatacattca aatatgtatc cgctcatgga
      121 tctcgatccc gcgaaattaa tacgactcac tatagggtat tttaacttta ctaaggagcg
      181 atcgcttccg aattcctacc gcggatatat gaagctctcc ctggtggccg cgatgctgct
      241 gctgctcagc gcggcgcggg ccgaggagga ggacaagaag gaggacgtgg gcacggtggt
      301 cggcatcgac ctggggacca cctactcctg cgtcggcgtg ttcaagaacg gccgcgtgga
      361 gatcatcgcc aacgatcagg gcaaccgcat cacgccgtcc tatgtcgcct tcactcctga
      421 aggggaacgt ctgattggcg atgccgccaa gaaccagctc acctccaacc ccgagaacac
      481 ggtctttgac gccaagcggc tcatcggccg cacgtggaat gacccgtctg tgcagcagga
      541 catcaagttc ttgccgttca aggtggttga aaagaaaact aaaccataca ttcaagttga
      601 tattggaggt gggcaaacaa agacatttgc tcctgaagaa atttctgcca tggttctcac
      661 taaaatgaaa gaaaccgctg aggcttattt gggaaagaag gttacccatg cagttgttac
      721 tgtaccagcc tattttaatg atgcccaacg ccaagcaacc aaagacgctg gaactattgc
      781 tggcctaaat gttatgagga tcatcaacga gcctacggca gctgctattg cttatggcct
      841 ggataagagg gagggggaga agaacatcct ggtgtttgac ctgggtggcg gaaccttcga
      901 tgtgtctctt ctcaccattg acaatggtgt cttcgaagtt gtggccacta atggagatac
      961 tcatctgggt ggagaagact ttgaccagcg tgtcatggaa cacttcatca aactgtacaa
     1021 aaagaagacg ggcaaagatg tcaggaaaga caatagagct gtgcagaaac tccggcgcga
     1081 ggtagaaaag gccaaacggg ccctgtcttc tcagcatcaa gcaagaattg aaattgagtc
     1141 cttctatgaa ggagaagact tttctgagac cctgactcgg gccaaatttg aagagctcaa
     1201 catggatctg ttccggtcta ctatgaagcc cgtccagaaa gtgttggaag attctgattt
     1261 gaagaagtct gatattgatg aaattgttct tgttggtggc tcgactcgaa ttccaaagat
     1321 tcagcaactg gttaaagagt tcttcaatgg caaggaacca tcccgtggca taaacccaga
     1381 tgaagctgta gcgtatggtg ctgctgtcca ggctggtgtg ctctctggtg atcaagatac
     1441 aggtgacctg gtactgcttg atgtatgtcc ccttacactt ggtattgaaa ctgtgggagg
     1501 tgtcatgacc aaactgattc caaggaacac agtggtgcct accaagaagt ctcagatctt
     1561 ttctacagct tctgataatc aaccaactgt tacaatcaag gtctatgaag gtgaaagacc
     1621 cctgacaaaa gacaatcatc ttctgggtac atttgatctg actggaattc ctcctgctcc
     1681 tcgtggggtc ccacagattg aagtcacctt tgagatagat gtgaatggta ttcttcgagt
     1741 gacagctgaa gacaagggta cagggaacaa aaataagatc acaatcacca atgaccagaa
     1801 tcgcctgaca cctgaagaaa tcgaaaggat ggttaatgat gctgagaagt ttgctgagga
     1861 agacaaaaag ctcaaggagc gcattgatac tagaaatgag ttggaaagct atgcctattc
     1921 tctaaagaat cagattggag ataaagaaaa gctgggaggt aaactttcct ctgaagataa
     1981 ggagaccatg gaaaaagctg tagaagaaaa gattgaatgg ctggaaagcc accaagatgc
     2041 tgacattgaa gacttcaaag ctaagaagaa ggaactggaa gaaattgttc aaccaattat
     2101 cagcaaactc tatggaagtg caggccctcc cccaactggt gaagaggata cagcagaaaa
     2161 agatgagttg gggccaacca ctgaggatct gtactttcag agcgataacg atggatccga
     2221 aatcggtact ggctttccat tcgaccccca ttatgtggaa gtcctgggcg agcgcatgca
     2281 ctacgtcgat gttggtccgc gcgatggcac ccctgtgctg ttcctgcacg gtaacccgac
     2341 ctcctcctac gtgtggcgca acatcatccc gcatgttgca ccgacccatc gctgcattgc
     2401 tccagacctg atcggtatgg gcaaatccga caaaccagac ctgggttatt tcttcgacga
     2461 ccacgtccgc ttcatggatg ccttcatcga agccctgggt ctggaagagg tcgtcctggt
     2521 cattcacgac tggggctccg ctctgggttt ccactgggcc aagcgcaatc cagagcgcgt
     2581 caaaggtatt gcatttatgg agttcatccg ccctatcccg acctgggacg aatggccaga
     2641 atttgcccgc gagaccttcc aggccttccg caccaccgac gtcggccgca agctgatcat
     2701 cgatcagaac gtttttatcg agggtacgct gccgatgggt gtcgtccgcc cgctgactga
     2761 agtcgagatg gaccattacc gcgagccgtt cctgaatcct gttgaccgcg agccactgtg
     2821 gcgcttccca aacgagctgc caatcgccgg tgagccagcg aacatcgtcg cgctggtcga
     2881 agaatacatg gactggctgc accagtcccc tgtcccgaag ctgctgttct ggggcacccc
     2941 aggcgttctg atcccaccgg ccgaagccgc tcgcctggcc aaaagcctgc ctaactgcaa
     3001 ggctgtggac atcggcccgg gtctgaatct gctgcaagaa gacaacccgg acctgatcgg
     3061 cagcgagatc gcgcgctggc tgtctactct ggagatttcc ggtcatcatc accatcacca
     3121 ctaatagtca accgctgagc aataactagc ataacccctt ggggcctcta aacgggtctt
     3181 gaggggtttt ttgctgaaag gaggaactat atccggttcg cttgctgtcc ataaaaccgc
     3241 ccagtctagc tatcgccatg taagcccact gcaagctacc tgctttctct ttgcgcttgc
     3301 gttttccctt gtccagatag cccagtagct gacattcatc cggggtcagc accgtttctg
     3361 cggactggct ttctacgtaa tggtttctta gacgtcaggt ggcacttttc ggggaaatgt
     3421 gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag
     3481 acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtatga gtattcaaca
     3541 tttccgtgtc gcccttattc ccttttttgc ggcattttgc cttcctgttt ttgctcaccc
     3601 agaaacgctg gtgaaagtaa aagatgctga agatcagttg ggtgcacgag tgggttacat
     3661 cgaactggat ctcaacagcg gtaagatcct tgagagtttt cgccccgaag aacgttttcc
     3721 aatgatgagc actttcaaag ttctgctatg tggcgcggta ttatcccgta ttgacgccgg
     3781 gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat gacttggttg agtactcacc
     3841 agtcacagaa aagcatctta cggatggcat gacagtaaga gaattatgca gtgctgccat
     3901 aaccatgagt gataacactg cggccaactt acttctgaca actatcggag gaccgaagga
     3961 gctaaccgct tttttgcaca acatggggga tcatgtaact cgccttgatc gttgggaacc
     4021 ggagctgaat gaagccatac caaacgacga gcgtgacacc acgatgcctg tagcaatggc
     4081 aacaacgttg cgcaaactat taactggcga actacttact ctagcttccc ggcaacaatt
     4141 aatagactgg atggaggcgg ataaagttgc aggaccactt ctgcgctcgg cccttccggc
     4201 tggctggttt attgctgata aatctggagc cggtgagcgt gggtctcgcg gtatcattgc
     4261 agcactgggg ccagatggta agccctcccg tatcgtagtt atctacacga cggggagtca
     4321 ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac tgattaagca
     4381 ttggtaattc gaaatgaccg accaagcgac gcccaaccgg tatcagctca ctcaaaggcg
     4441 gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc
     4501 cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc
     4561 ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga
     4621 ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc
     4681 ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat
     4741 agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg
     4801 cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc
     4861 aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga
     4921 gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact
     4981 agaaggacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt
     5041 ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag
     5101 cagcagatta cgcgcagaaa aaaaggattt caagaagatc ctttgatctt ttctacgggg
     5161 tctgacgctc agtggaacga aaactcacgt taagggcata tgatggtagt gtggggtctc
     5221 cccatgcgag agtagggaac tgccaggcat caaataaaac gaaaggctca gtcgaaagac
     5281 tgggcctttc gttttatctg ttgtttgtcg gtgaacgctc tcctgagtag gacaaatccg
     5341 ccgggagcgg atttgaacgt tgcgaagcaa cggcccggag ggtggcgggc aggacgcccg
//
