LOCUS       sequence-345555-        5057 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 5057)
  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 5057)
  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_PKM-Halo
FEATURES             Location/Qualifiers
     source          1..5057
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      32..59
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     primer_bind     141..160
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        141..159
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      199..208
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1843..1863
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="EDLYFQS"
     CDS             1879..2763
                     /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             2764..2781
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(2792..2810)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      2806..2853
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        3082..3186
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3187..3255,3256..4047)
                     /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(3405..3424)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      complement(4205..4793)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4285..4304)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      4911..4997
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
ORIGIN
        1 ccgccataaa ctgccaggca tcaaattaag cagaaggcca tcctgacgga tggccttttt
       61 gcgtttctac aaactctttt gtttattttt ctaaatacat tcaaatatgt atccgctcat
      121 ggatctcgat cccgcgaaat taatacgact cactataggg tattttaact ttactaagga
      181 gcgatcgctt ccgaattcgc caccatggcg tcgaagcccc atagtgaagc cgggactgcc
      241 ttcattcaga cccagcagct gcacgcagcc atggctgaca cattcctgga gcacatgtgc
      301 cgcctggaca ttgattcacc acccatcaca gcccggaaca ctggcatcat ctgtaccatt
      361 ggcccagctt cccgatcagt ggagacgttg aaggagatga ttaagtctgg aatgaatgtg
      421 gctcgtctga acttctctca tggaactcat gagtaccatg cggagaccat caagaatgtg
      481 cgcacagcca cggaaagctt tgcttctgac cccatcctct accggcccgt tgctgtggct
      541 ctagacacta aaggacctga gatccgaact gggctcatca agggcagcgg cactgcagag
      601 gtggagctga agaagggagc cactctcaaa atcacgctgg ataacgccta catggaaaag
      661 tgtgacgaga acatcctgtg gctggactac aagaacatct gcaaggtggt ggaagtgggc
      721 agcaagatct acgtggatga tgggcttatt tctctccagg tgaagcagaa aggtgccgac
      781 ttcctggtga cggaggtgga aaatggtggc tccttgggca gcaagaaggg tgtgaacctt
      841 cctggggctg ctgtggactt gcctgctgtg tcggagaagg acatccagga tctgaagttt
      901 ggggtcgagc aggatgttga tatggtgttt gcgtcattca tccgcaaggc atctgatgtc
      961 catgaagtta ggaaggtcct gggagagaag ggaaagaaca tcaagattat cagcaaaatc
     1021 gagaatcatg agggggttcg gaggtttgat gaaatcctgg aggccagtga tgggatcatg
     1081 gtggctcgtg gtgatctagg cattgagatt cctgcagaga aggtcttcct tgctcagaag
     1141 atgatgattg gacggtgcaa ccgagctggg aagcctgtca tctgtgctac tcagatgctg
     1201 gagagcatga tcaagaagcc ccgccccact cgggctgaag gcagtgatgt ggccaatgca
     1261 gtcctggatg gagccgactg catcatgctg tctggagaaa cagccaaagg ggactatcct
     1321 ctggaggctg tgcgcatgca gcacctgatt gcccgtgagg cagaggctgc catctaccac
     1381 ttgcaattat ttgaggaact ccgccgcctg gcgcccatta ccagcgaccc cacagaagcc
     1441 accgccgtgg gtgccgtgga ggcctccttc aagtgctgca gtggggccat aatcgtcctc
     1501 accaagtctg gcaggtctgc tcaccaggtg gccagatacc gcccacgtgc ccccatcatt
     1561 gctgtgaccc ggaatcccca gacagctcgt caggcccacc tgtaccgtgg catcttccct
     1621 gtgctgtgca aggacccagt ccaggaggcc tgggctgagg acgtggacct ccgggtgaac
     1681 tttgccatga atgttggcaa ggcccgaggc ttcttcaaga agggagatgt ggtcattgtg
     1741 ctgaccggat ggcgccctgg ctccggcttc accaacacca tgcgtgttgt tcctgtgccg
     1801 ggcggctccg gctcctgcag gcgagctctc gagccaacca ctgaggatct gtactttcag
     1861 agcgataacg atggatccga aatcggtact ggctttccat tcgaccccca ttatgtggaa
     1921 gtcctgggcg agcgcatgca ctacgtcgat gttggtccgc gcgatggcac ccctgtgctg
     1981 ttcctgcacg gtaacccgac ctcctcctac gtgtggcgca acatcatccc gcatgttgca
     2041 ccgacccatc gctgcattgc tccagacctg atcggtatgg gcaaatccga caaaccagac
     2101 ctgggttatt tcttcgacga ccacgtccgc ttcatggatg ccttcatcga agccctgggt
     2161 ctggaagagg tcgtcctggt cattcacgac tggggctccg ctctgggttt ccactgggcc
     2221 aagcgcaatc cagagcgcgt caaaggtatt gcatttatgg agttcatccg ccctatcccg
     2281 acctgggacg aatggccaga atttgcccgc gagaccttcc aggccttccg caccaccgac
     2341 gtcggccgca agctgatcat cgatcagaac gtttttatcg agggtacgct gccgatgggt
     2401 gtcgtccgcc cgctgactga agtcgagatg gaccattacc gcgagccgtt cctgaatcct
     2461 gttgaccgcg agccactgtg gcgcttccca aacgagctgc caatcgccgg tgagccagcg
     2521 aacatcgtcg cgctggtcga agaatacatg gactggctgc accagtcccc tgtcccgaag
     2581 ctgctgttct ggggcacccc aggcgttctg atcccaccgg ccgaagccgc tcgcctggcc
     2641 aaaagcctgc ctaactgcaa ggctgtggac atcggcccgg gtctgaatct gctgcaagaa
     2701 gacaacccgg acctgatcgg cagcgagatc gcgcgctggc tgtctactct ggagatttcc
     2761 ggtcatcatc accatcacca ctaatagtca accgctgagc aataactagc ataacccctt
     2821 ggggcctcta aacgggtctt gaggggtttt ttgctgaaag gaggaactat atccggttcg
     2881 cttgctgtcc ataaaaccgc ccagtctagc tatcgccatg taagcccact gcaagctacc
     2941 tgctttctct ttgcgcttgc gttttccctt gtccagatag cccagtagct gacattcatc
     3001 cggggtcagc accgtttctg cggactggct ttctacgtaa tggtttctta gacgtcaggt
     3061 ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca
     3121 aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg
     3181 aagagtatga gtattcaaca tttccgtgtc gcccttattc ccttttttgc ggcattttgc
     3241 cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga agatcagttg
     3301 ggtgcacgag tgggttacat cgaactggat ctcaacagcg gtaagatcct tgagagtttt
     3361 cgccccgaag aacgttttcc aatgatgagc actttcaaag ttctgctatg tggcgcggta
     3421 ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat
     3481 gacttggttg agtactcacc agtcacagaa aagcatctta cggatggcat gacagtaaga
     3541 gaattatgca gtgctgccat aaccatgagt gataacactg cggccaactt acttctgaca
     3601 actatcggag gaccgaagga gctaaccgct tttttgcaca acatggggga tcatgtaact
     3661 cgccttgatc gttgggaacc ggagctgaat gaagccatac caaacgacga gcgtgacacc
     3721 acgatgcctg tagcaatggc aacaacgttg cgcaaactat taactggcga actacttact
     3781 ctagcttccc ggcaacaatt aatagactgg atggaggcgg ataaagttgc aggaccactt
     3841 ctgcgctcgg cccttccggc tggctggttt attgctgata aatctggagc cggtgagcgt
     3901 gggtctcgcg gtatcattgc agcactgggg ccagatggta agccctcccg tatcgtagtt
     3961 atctacacga cggggagtca ggcaactatg gatgaacgaa atagacagat cgctgagata
     4021 ggtgcctcac tgattaagca ttggtaattc gaaatgaccg accaagcgac gcccaaccgg
     4081 tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa
     4141 agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg
     4201 cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga
     4261 ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg
     4321 tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg
     4381 gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc
     4441 gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg
     4501 gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca
     4561 ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt
     4621 ggcctaacta cggctacact agaaggacag tatttggtat ctgcgctctg ctgaagccag
     4681 ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg
     4741 gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggattt caagaagatc
     4801 ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggcata
     4861 tgatggtagt gtggggtctc cccatgcgag agtagggaac tgccaggcat caaataaaac
     4921 gaaaggctca gtcgaaagac tgggcctttc gttttatctg ttgtttgtcg gtgaacgctc
     4981 tcctgagtag gacaaatccg ccgggagcgg atttgaacgt tgcgaagcaa cggcccggag
     5041 ggtggcgggc aggacgc
//
