LOCUS       sequence-345557-        5074 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 5074)
  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 5074)
  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_CCT3-Halo
FEATURES             Location/Qualifiers
     source          1..5074
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      193..279
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      371..398
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     primer_bind     480..499
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        480..498
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      556..565
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2239..2259
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="EDLYFQS"
     CDS             2275..3159
                     /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             3160..3177
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(3188..3206)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      3202..3249
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        3478..3582
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3583..3651,3652..4443)
                     /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(3801..3820)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      complement(join(4601..5074,1..115))
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4681..4700)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag
       61 cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ttcaagaaga
      121 tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaacttca cgttaagggc
      181 atagccaggc atcaaataaa acgaaaggct cagtcgaaag actgggcctt tcgttttatc
      241 tgttgtttgt cggtgaacgc tctcctgagt aggacaaatc cgccgggagc ggatttgaac
      301 gttgcgaagc aacggcccgg agggtggcgg gcaggacgcc cgccataaac tgccaggcat
      361 caaattaagc agaaggccat cctgacggat ggcctttttg cgtttctaca aactcttttg
      421 tttatttttc taaatacatt caaatatgta tccgctcatg gatctcgatc ccgcgaaatt
      481 aatacgactc actatagggt attttaactt tactaaggag cgatcgcttc cgaattccta
      541 ccgcggatat ctagagccac catggcgggc catcgtccag tgctcgtgct cagccagaac
      601 acaaagcgtg aatccggaag aaaagttcaa tctggaaaca tcaatgctgc caagactatt
      661 gcagatatca tccgaacatg tttgggaccc aagtccatga tgaagatgct tttggaccca
      721 atgggaggca ttgtgatgac caatgatggc aatgccattc ttcgagagat tcaagtccag
      781 catccagcgg ccaagtccat gatcgaaatt agccggaccc aggatgaaga ggttggagat
      841 gggaccacat cagtaattat tcttgcaggg gaaatgctgt ctgtagctga gcacttcctg
      901 gagcagcaga tgcacccaac agtggtgatc agtgcttacc gcaaggcatt ggatgatatg
      961 atcagcaccc taaagaaaat aagtatccca gtcgacatca gtgacagtga tatgatgctg
     1021 aacatcatca acagctctat tactaccaaa gccatcagtc ggtggtcatc tttggcttgc
     1081 aacattgccc tggatgctgt caagatggta cagtttgagg agaatggtcg gaaagagatt
     1141 gacataaaaa aatatgcaag agtggaaaag atacctggag gcatcattga agactcctgt
     1201 gtcttgcgtg gagtcatgat taacaaggat gtgacccatc cacgtatgcg gcgctatatc
     1261 aagaaccctc gcattgtgct gctggattct tctctggaat acaagaaagg agaaagccag
     1321 actgacattg agattacacg agaggaggac ttcacccgaa ttctccagat ggaggaagag
     1381 tacatccagc agctctgtga ggacattatc caactgaagc ccgatgtggt catcactgaa
     1441 aagggcatct cagatttagc tcagcactac cttatgcggg ccaatatcac agccatccgc
     1501 agagtccgga agacagacaa taatcgcatt gctagagcct gtggggcccg gatagtcagc
     1561 cgaccagagg aactgagaga agatgatgtt ggaacaggag caggcctgtt ggaaatcaag
     1621 aaaattggag atgaatactt tactttcatc actgactgca aagaccccaa ggcctgcacc
     1681 attctcctcc ggggggctag caaagagatt ctctcggaag tagaacgcaa cctccaggat
     1741 gccatgcaag tgtgtcgcaa tgttctcctg gaccctcagc tggtgccagg gggtggggcc
     1801 tccgagatgg ctgtggccca tgccttgaca gaaaaatcca aggccatgac tggtgtggaa
     1861 caatggccat acagggctgt tgcccaggcc ctagaggtca ttcctcgtac cctgatccag
     1921 aactgtgggg ccagcaccat ccgtctactt acctcccttc gggccaagca cacccaggag
     1981 aactgtgaga cctggggtgt aaatggtgag acgggtactt tggtggacat gaaggaactg
     2041 ggcatatggg agccattggc tgtgaagctg cagacttata agacagcagt ggagacggca
     2101 gttctgctac tgcgaattga tgacatcgtt tcaggccaca aaaagaaagg cgatgaccag
     2161 agccggcaag gcggggctcc tgatgctggc caggagggcg gctccggctc ctgcaggcga
     2221 gctctcgagc caaccactga ggatctgtac tttcagagcg ataacgatgg atccgaaatc
     2281 ggtactggct ttccattcga cccccattat gtggaagtcc tgggcgagcg catgcactac
     2341 gtcgatgttg gtccgcgcga tggcacccct gtgctgttcc tgcacggtaa cccgacctcc
     2401 tcctacgtgt ggcgcaacat catcccgcat gttgcaccga cccatcgctg cattgctcca
     2461 gacctgatcg gtatgggcaa atccgacaaa ccagacctgg gttatttctt cgacgaccac
     2521 gtccgcttca tggatgcctt catcgaagcc ctgggtctgg aagaggtcgt cctggtcatt
     2581 cacgactggg gctccgctct gggtttccac tgggccaagc gcaatccaga gcgcgtcaaa
     2641 ggtattgcat ttatggagtt catccgccct atcccgacct gggacgaatg gccagaattt
     2701 gcccgcgaga ccttccaggc cttccgcacc accgacgtcg gccgcaagct gatcatcgat
     2761 cagaacgttt ttatcgaggg tacgctgccg atgggtgtcg tccgcccgct gactgaagtc
     2821 gagatggacc attaccgcga gccgttcctg aatcctgttg accgcgagcc actgtggcgc
     2881 ttcccaaacg agctgccaat cgccggtgag ccagcgaaca tcgtcgcgct ggtcgaagaa
     2941 tacatggact ggctgcacca gtcccctgtc ccgaagctgc tgttctgggg caccccaggc
     3001 gttctgatcc caccggccga agccgctcgc ctggccaaaa gcctgcctaa ctgcaaggct
     3061 gtggacatcg gcccgggtct gaatctgctg caagaagaca acccggacct gatcggcagc
     3121 gagatcgcgc gctggctgtc tactctggag atttccggtc atcatcacca tcaccactaa
     3181 tagtcaaccg ctgagcaata actagcataa ccccttgggg cctctaaacg ggtcttgagg
     3241 ggttttttgc tgaaaggagg aactatatcc ggttcgcttg ctgtccataa aaccgcccag
     3301 tctagctatc gccatgtaag cccactgcaa gctacctgct ttctctttgc gcttgcgttt
     3361 tcccttgtcc agatagccca gtagctgaca ttcatccggg gtcagcaccg tttctgcgga
     3421 ctggctttct acgtaatggt ttcttagacg tcaggtggca cttttcgggg aaatgtgcgc
     3481 ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa
     3541 taaccctgat aaatgcttca ataatattga aaaaggaaga gtatgagtat tcaacatttc
     3601 cgtgtcgccc ttattccctt ttttgcggca ttttgccttc ctgtttttgc tcacccagaa
     3661 acgctggtga aagtaaaaga tgctgaagat cagttgggtg cacgagtggg ttacatcgaa
     3721 ctggatctca acagcggtaa gatccttgag agttttcgcc ccgaagaacg ttttccaatg
     3781 atgagcactt tcaaagttct gctatgtggc gcggtattat cccgtattga cgccgggcaa
     3841 gagcaactcg gtcgccgcat acactattct cagaatgact tggttgagta ctcaccagtc
     3901 acagaaaagc atcttacgga tggcatgaca gtaagagaat tatgcagtgc tgccataacc
     3961 atgagtgata acactgcggc caacttactt ctgacaacta tcggaggacc gaaggagcta
     4021 accgcttttt tgcacaacat gggggatcat gtaactcgcc ttgatcgttg ggaaccggag
     4081 ctgaatgaag ccataccaaa cgacgagcgt gacaccacga tgcctgtagc aatggcaaca
     4141 acgttgcgca aactattaac tggcgaacta cttactctag cttcccggca acaattaata
     4201 gactggatgg aggcggataa agttgcagga ccacttctgc gctcggccct tccggctggc
     4261 tggtttattg ctgataaatc tggagccggt gagcgtgggt ctcgcggtat cattgcagca
     4321 ctggggccag atggtaagcc ctcccgtatc gtagttatct acacgacggg gagtcaggca
     4381 actatggatg aacgaaatag acagatcgct gagataggtg cctcactgat taagcattgg
     4441 taattcgaaa tgaccgacca agcgacgccc aaccggtatc agctcactca aaggcggtaa
     4501 tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc
     4561 aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc
     4621 ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat
     4681 aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc
     4741 cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct
     4801 cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg
     4861 aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc
     4921 cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga
     4981 ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa
     5041 ggacagtatt tggtatctgc gctctgctga agcc
//
