LOCUS       sequence-345594-        5235 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 5235)
  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 5235)
  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_ABCE1-Halo
FEATURES             Location/Qualifiers
     source          1..5235
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      290..376
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      468..495
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     primer_bind     577..596
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        577..595
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             2457..2477
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="EDLYFQS"
     CDS             2493..3377
                     /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             3378..3395
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(3406..3424)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      3420..3467
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        3696..3800
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3801..3869,3870..4661)
                     /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(4019..4038)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      complement(join(4819..5235,1..172))
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4899..4918)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gcctaactac ggctacacta gaaggacagt atttggtatc tgcgctctgc tgaagccagt
       61 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg
      121 tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatttc aagaagatcc
      181 tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggcatat
      241 gatggtagtg tggggtctcc ccatgcgaga gtagggaact gccaggcatc aaataaaacg
      301 aaaggctcag tcgaaagact gggcctttcg ttttatctgt tgtttgtcgg tgaacgctct
      361 cctgagtagg acaaatccgc cgggagcgga tttgaacgtt gcgaagcaac ggcccggagg
      421 gtggcgggca ggacgcccgc cataaactgc caggcatcaa attaagcaga aggccatcct
      481 gacggatggc ctttttgcgt ttctacaaac tcttttgttt atttttctaa atacattcaa
      541 atatgtatcc gctcatggat ctcgatcccg cgaaattaat acgactcact atagggtatt
      601 ttaactttac taaggagcga tcgcttccga attcctaccg cggatatatg gcagacaagt
      661 taacgagaat tgctattgtc aaccatgaca aatgtaaacc taagaaatgt cgacaggaat
      721 gcaaaaagag ttgtcctgta gttcgaatgg gaaaattatg catagaggtt acaccccaga
      781 gcaaaatagc atggatttcc gaaactcttt gtattggttg tggtatctgt attaagaaat
      841 gcccctttgg cgccttatca attgtcaatc taccaagcaa cttggaaaaa gaaaccacac
      901 atcgatattg tgccaatgcc ttcaaacttc acaggttgcc tatccctcgt ccaggtgaag
      961 ttttgggatt agttggaact aatggtattg gaaagtcaac tgctttaaaa attttagcag
     1021 gaaaacaaaa gccaaacctt ggaaagtacg atgatcctcc tgactggcag gagattttga
     1081 cttatttccg tggatctgaa ttacaaaatt actttacaaa gattctagaa gatgacctaa
     1141 aagccatcat caaacctcaa tatgtagacc agattcctaa ggctgcaaag gggacagtgg
     1201 gatctatttt ggaccgaaaa gatgaaacaa agacacaggc aattgtatgt cagcagcttg
     1261 atttaaccca cctaaaagaa cgaaatgttg aagatctttc aggaggagag ttgcagagat
     1321 ttgcttgtgc tgtcgtttgc atacagaaag ctgatatttt catgtttgat gagccttcta
     1381 gttacctaga tgtcaagcag cgtttaaagg ctgctattac tatacgatct ctaataaatc
     1441 cagatagata tatcattgtg gtggaacatg atctaagtgt attagactat ctctccgact
     1501 tcatctgctg tttatatggt gtaccaagcg cctatggagt tgtcactatg ccttttagtg
     1561 taagagaagg cataaacatt tttttggatg gctatgttcc aacagaaaac ttgagattca
     1621 gagatgcatc acttgttttt aaagtggctg agacagcaaa tgaagaagaa gttaaaaaga
     1681 tgtgtatgta taaatatcca ggaatgaaga aaaaaatggg agaatttgag ctagcaattg
     1741 tagctggaga gtttacagat tctgaaatta tggtgatgct gggggaaaat ggaacgggta
     1801 aaacgacatt tatcagaatg cttgctggaa gacttaaacc tgatgaagga ggagaagtac
     1861 cagttctaaa tgtcagttat aagccacaga aaattagtcc caaatcaact ggaagtgttc
     1921 gccagttact acatgaaaag ataagagatg cttatactca cccacaattt gtgaccgatg
     1981 taatgaagcc tctgcaaatt gaaaacatca ttgatcaaga ggtgcagaca ttatctggtg
     2041 gtgaactaca gcgagtagct ttagcccttt gcttgggcaa acctgctgat gtctatttaa
     2101 ttgatgaacc atctgcatat ttggattctg agcaaagact gatggcagct cgagttgtca
     2161 aacgtttcat actccatgca aaaaagacag cctttgttgt ggaacatgac ttcatcatgg
     2221 ccacctatct agcggatcgc gtcatcgttt ttgatggtgt tccatctaag aacacagttg
     2281 caaacagtcc tcaaaccctt ttggctggca tgaataaatt tttgtctcag cttgaaatta
     2341 cattcagaag agatccaaac aactataggc cacgaataaa caaacttaat tcaattaagg
     2401 atgtagaaca aaagaagagt ggaaactact ttttcttgga tgatgggcca accactgagg
     2461 atctgtactt tcagagcgat aacgatggat ccgaaatcgg tactggcttt ccattcgacc
     2521 cccattatgt ggaagtcctg ggcgagcgca tgcactacgt cgatgttggt ccgcgcgatg
     2581 gcacccctgt gctgttcctg cacggtaacc cgacctcctc ctacgtgtgg cgcaacatca
     2641 tcccgcatgt tgcaccgacc catcgctgca ttgctccaga cctgatcggt atgggcaaat
     2701 ccgacaaacc agacctgggt tatttcttcg acgaccacgt ccgcttcatg gatgccttca
     2761 tcgaagccct gggtctggaa gaggtcgtcc tggtcattca cgactggggc tccgctctgg
     2821 gtttccactg ggccaagcgc aatccagagc gcgtcaaagg tattgcattt atggagttca
     2881 tccgccctat cccgacctgg gacgaatggc cagaatttgc ccgcgagacc ttccaggcct
     2941 tccgcaccac cgacgtcggc cgcaagctga tcatcgatca gaacgttttt atcgagggta
     3001 cgctgccgat gggtgtcgtc cgcccgctga ctgaagtcga gatggaccat taccgcgagc
     3061 cgttcctgaa tcctgttgac cgcgagccac tgtggcgctt cccaaacgag ctgccaatcg
     3121 ccggtgagcc agcgaacatc gtcgcgctgg tcgaagaata catggactgg ctgcaccagt
     3181 cccctgtccc gaagctgctg ttctggggca ccccaggcgt tctgatccca ccggccgaag
     3241 ccgctcgcct ggccaaaagc ctgcctaact gcaaggctgt ggacatcggc ccgggtctga
     3301 atctgctgca agaagacaac ccggacctga tcggcagcga gatcgcgcgc tggctgtcta
     3361 ctctggagat ttccggtcat catcaccatc accactaata gtcaaccgct gagcaataac
     3421 tagcataacc ccttggggcc tctaaacggg tcttgagggg ttttttgctg aaaggaggaa
     3481 ctatatccgg ttcgcttgct gtccataaaa ccgcccagtc tagctatcgc catgtaagcc
     3541 cactgcaagc tacctgcttt ctctttgcgc ttgcgttttc ccttgtccag atagcccagt
     3601 agctgacatt catccggggt cagcaccgtt tctgcggact ggctttctac gtaatggttt
     3661 cttagacgtc aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt
     3721 tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat
     3781 aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt
     3841 ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg
     3901 ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga
     3961 tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttc aaagttctgc
     4021 tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga gcaactcggt cgccgcatac
     4081 actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg
     4141 gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca
     4201 acttacttct gacaactatc ggaggaccga aggagctaac cgcttttttg cacaacatgg
     4261 gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg
     4321 acgagcgtga caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg
     4381 gcgaactact tactctagct tcccggcaac aattaataga ctggatggag gcggataaag
     4441 ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg
     4501 gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct
     4561 cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac
     4621 agatcgctga gataggtgcc tcactgatta agcattggta attcgaaatg accgaccaag
     4681 cgacgcccaa ccggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg
     4741 ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa
     4801 ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg
     4861 acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc
     4921 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc
     4981 ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc
     5041 ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg
     5101 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc
     5161 actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga
     5221 gttcttgaag tggtg
//
