LOCUS       sequence-349802-        4460 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Production of HaloTag fusion protein.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4460)
  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 4460)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 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_xtHUR-Halo
FEATURES             Location/Qualifiers
     source          1..4460
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      11..38
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     primer_bind     120..139
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        120..138
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      196..205
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1225..1245
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="EDLYFQS"
     CDS             1261..2145
                     /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             2146..2163
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(2174..2192)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      2188..2235
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        2464..2568
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2569..2637,2638..3429)
                     /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(2787..2806)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      complement(3587..4175)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3667..3686)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      4293..4379
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
ORIGIN
        1 caaattaagc agaaggccat cctgacggat ggcctttttg cgtttctaca aactcttttg
       61 tttatttttc taaatacatt caaatatgta tccgctcatg gatctcgatc ccgcgaaatt
      121 aatacgactc actatagggt attttaactt tactaaggag cgatcgcttc cgaattccta
      181 ccgcggatat ctagagccac catggcgtct aacggttatg gagaccacat ggatgacgtg
      241 tgcagggatg acattggtag gaccaatcta attgtgaact acctgcctca gaatatgaca
      301 caagatgaac tgcgaagcct tttcagcagt atcggagagg tagaatctgc aaagcttatt
      361 cgggataaag ttgcagggca cagtttaggc tatggttttg tgaattacct taatgcaaaa
      421 gatgcagaaa gagcaataaa cactcttaat ggcctgaggc tgcaatctaa aactataaag
      481 gtctccgttg ctcgtccaag ttcagaatcc attaaagatg ctaatttgta catcagcggg
      541 ctcccaagga caatgacaca aaaagatgta gaagatatgt ttttaccttt tgggcgtatt
      601 atcaattcaa gggtgctggt tgatcaggca acaggtttgt cccggggagt tgcctttata
      661 cggtttgata aaagatccga agctgaagag gccattgcca gtttcaatgg acacaaacca
      721 cctggatctt cagagcctat tacagtaaaa tttgctgcca atccaaatca gaacaagaat
      781 atggcccttc tttcacagct ttgccactct ccagctcgac gatttggagg gcctgtccat
      841 caccaggcac aaaggttcag gttttctcca atgggtgtag atcacatgag cagcatatcc
      901 ggtgtaaatg ttgcaagcag tgcatcttct ggttggtgca tctttatcta caatcttggc
      961 caagatgccg atgaaggaat cctttggcaa atgtttgggc cttttggagc agtcactaat
     1021 gtcaaggtta ttcgtgattt taacaccaac aaatgtaaag gttttggttt tgtgaccatg
     1081 acaaactatg aagaagcagc aatggctatc gcaagtctta atggctaccg tctaggggac
     1141 aaaaccttac aagttttctt caaaaccagc aagtcacaca aaggcggctc cggctcctgc
     1201 aggcgagctc tcgagccaac cactgaggat ctgtactttc agagcgataa cgatggatcc
     1261 gaaatcggta ctggctttcc attcgacccc cattatgtgg aagtcctggg cgagcgcatg
     1321 cactacgtcg atgttggtcc gcgcgatggc acccctgtgc tgttcctgca cggtaacccg
     1381 acctcctcct acgtgtggcg caacatcatc ccgcatgttg caccgaccca tcgctgcatt
     1441 gctccagacc tgatcggtat gggcaaatcc gacaaaccag acctgggtta tttcttcgac
     1501 gaccacgtcc gcttcatgga tgccttcatc gaagccctgg gtctggaaga ggtcgtcctg
     1561 gtcattcacg actggggctc cgctctgggt ttccactggg ccaagcgcaa tccagagcgc
     1621 gtcaaaggta ttgcatttat ggagttcatc cgccctatcc cgacctggga cgaatggcca
     1681 gaatttgccc gcgagacctt ccaggccttc cgcaccaccg acgtcggccg caagctgatc
     1741 atcgatcaga acgtttttat cgagggtacg ctgccgatgg gtgtcgtccg cccgctgact
     1801 gaagtcgaga tggaccatta ccgcgagccg ttcctgaatc ctgttgaccg cgagccactg
     1861 tggcgcttcc caaacgagct gccaatcgcc ggtgagccag cgaacatcgt cgcgctggtc
     1921 gaagaataca tggactggct gcaccagtcc cctgtcccga agctgctgtt ctggggcacc
     1981 ccaggcgttc tgatcccacc ggccgaagcc gctcgcctgg ccaaaagcct gcctaactgc
     2041 aaggctgtgg acatcggccc gggtctgaat ctgctgcaag aagacaaccc ggacctgatc
     2101 ggcagcgaga tcgcgcgctg gctgtctact ctggagattt ccggtcatca tcaccatcac
     2161 cactaatagt caaccgctga gcaataacta gcataacccc ttggggcctc taaacgggtc
     2221 ttgaggggtt ttttgctgaa aggaggaact atatccggtt cgcttgctgt ccataaaacc
     2281 gcccagtcta gctatcgcca tgtaagccca ctgcaagcta cctgctttct ctttgcgctt
     2341 gcgttttccc ttgtccagat agcccagtag ctgacattca tccggggtca gcaccgtttc
     2401 tgcggactgg ctttctacgt aatggtttct tagacgtcag gtggcacttt tcggggaaat
     2461 gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg
     2521 agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtat gagtattcaa
     2581 catttccgtg tcgcccttat tccctttttt gcggcatttt gccttcctgt ttttgctcac
     2641 ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg agtgggttac
     2701 atcgaactgg atctcaacag cggtaagatc cttgagagtt ttcgccccga agaacgtttt
     2761 ccaatgatga gcactttcaa agttctgcta tgtggcgcgg tattatcccg tattgacgcc
     2821 gggcaagagc aactcggtcg ccgcatacac tattctcaga atgacttggt tgagtactca
     2881 ccagtcacag aaaagcatct tacggatggc atgacagtaa gagaattatg cagtgctgcc
     2941 ataaccatga gtgataacac tgcggccaac ttacttctga caactatcgg aggaccgaag
     3001 gagctaaccg cttttttgca caacatgggg gatcatgtaa ctcgccttga tcgttgggaa
     3061 ccggagctga atgaagccat accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg
     3121 gcaacaacgt tgcgcaaact attaactggc gaactactta ctctagcttc ccggcaacaa
     3181 ttaatagact ggatggaggc ggataaagtt gcaggaccac ttctgcgctc ggcccttccg
     3241 gctggctggt ttattgctga taaatctgga gccggtgagc gtgggtctcg cggtatcatt
     3301 gcagcactgg ggccagatgg taagccctcc cgtatcgtag ttatctacac gacggggagt
     3361 caggcaacta tggatgaacg aaatagacag atcgctgaga taggtgcctc actgattaag
     3421 cattggtaat tcgaaatgac cgaccaagcg acgcccaacc ggtatcagct cactcaaagg
     3481 cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag
     3541 gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc
     3601 gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag
     3661 gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga
     3721 ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc
     3781 atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg
     3841 tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt
     3901 ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca
     3961 gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca
     4021 ctagaaggac agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag
     4081 ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca
     4141 agcagcagat tacgcgcaga aaaaaaggat ttcaagaaga tcctttgatc ttttctacgg
     4201 ggtctgacgc tcagtggaac gaaaactcac gttaagggca tatgatggta gtgtggggtc
     4261 tccccatgcg agagtaggga actgccaggc atcaaataaa acgaaaggct cagtcgaaag
     4321 actgggcctt tcgttttatc tgttgtttgt cggtgaacgc tctcctgagt aggacaaatc
     4381 cgccgggagc ggatttgaac gttgcgaagc aacggcccgg agggtggcgg gcaggacgcc
     4441 cgccataaac tgccaggcat
//
