LOCUS       sequence-345514-        5213 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 5213)
  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 5213)
  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_IGF2BP1_R167H-Halo
FEATURES             Location/Qualifiers
     source          1..5213
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      25..52
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     primer_bind     134..153
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        134..152
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      210..219
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1992..2012
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="EDLYFQS"
     CDS             2028..2912
                     /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             2913..2930
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(2941..2959)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      2955..3002
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        3231..3335
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3336..3404,3405..4196)
                     /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(3554..3573)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      complement(4354..4942)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4434..4453)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      5060..5146
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
ORIGIN
        1 aaactgccag gcatcaaatt aagcagaagg ccatcctgac ggatggcctt tttgcgtttc
       61 tacaaactct tttgtttatt tttctaaata cattcaaata tgtatccgct catggatctc
      121 gatcccgcga aattaatacg actcactata gggtatttta actttactaa ggagcgatcg
      181 cttccgaatt cctaccgcgg atatctagag ccaccatggc gaacaagctt tacatcggca
      241 acctcaacga gagcgtgacc cccgcggact tggagaaagt gtttgcggag cacaagatct
      301 cctacagcgg ccagttcttg gtcaaatccg gctacgcctt cgtggactgc ccggacgagc
      361 actgggcgat gaaggccatc gaaactttct ccgggaaagt agaattacaa ggaaaacgct
      421 tagagattga acattcggtg cccaaaaaac aaaggagccg gaaaattcaa atccgaaata
      481 ttccacccca gctccgatgg gaagtactgg acagcctgct ggctcagtat ggtacagtag
      541 agaactgtga gcaagtgaac accgagagtg agacggcagt ggtgaatgtc acctattcca
      601 accgggagca gaccaggcaa gccatcatga agctgaatgg ccaccagttg gagaaccatg
      661 ccctgaaggt ctcctacatc cccgatgagc agatagcaca gggacctgag aatgggcacc
      721 gagggggctt tggctctcgg ggtcagcccc gccagggctc acctgtggca gcgggggccc
      781 cagccaagca gcagcaagtg gacatccccc ttcggctcct ggtgcccacc cagtatgtgg
      841 gtgccattat tggcaaggag ggggccacca tccgcaacat cacaaaacag acccagtcca
      901 agatagacgt gcataggaag gagaacgcag gtgcagctga aaaagccatc agtgtgcact
      961 ccacccctga gggctgctcc tccgcttgta agatgatctt ggagattatg cataaagagg
     1021 ctaaggacac caaaacggct gacgaggttc ccctgaagat cctggcccat aataactttg
     1081 tagggcgtct cattggcaag gaaggacgga acctgaagaa ggtagagcaa gataccgaga
     1141 caaaaatcac catctcctcg ttgcaagacc ttacccttta caaccctgag aggaccatca
     1201 ctgtgaaggg ggccatcgag aattgttgca gggccgagca ggaaataatg aagaaagttc
     1261 gggaggccta tgagaatgat gtggctgcca tgagcctgca gtctcacctg atccctggcc
     1321 tgaacctggc tgctgtaggt cttttcccag cttcatccag cgcagtcccg ccgcctccca
     1381 gcagcgttac tggggctgct ccctatagct cctttatgca ggctcccgag caggagatgg
     1441 tgcaggtgtt tatccccgcc caggcagtgg gcgccatcat cggcaagaag gggcagcaca
     1501 tcaaacagct ctcccggttt gccagcgcct ccatcaagat tgcaccaccc gaaacacctg
     1561 actccaaagt tcgtatggtt atcatcactg gaccgccaga ggcccaattc aaggctcagg
     1621 gaagaatcta tggcaaactc aaggaggaga acttctttgg tcccaaggag gaagtgaagc
     1681 tggagaccca catacgtgtg ccagcatcag cagctggccg ggtcattggc aaaggtggaa
     1741 aaacggtgaa cgagttgcag aatttgacgg cagctgaggt ggtagtacca agagaccaga
     1801 cccctgatga gaacgaccag gtcatcgtga aaatcatcgg acatttctat gccagtcaga
     1861 tggctcaacg gaagatccga gacatcctgg cccaggttaa gcagcagcat cagaagggac
     1921 agagtaacca ggcccaggca cggaggaagg gcggctccgg ctcctgcagg cgagctctcg
     1981 agccaaccac tgaggatctg tactttcaga gcgataacga tggatccgaa atcggtactg
     2041 gctttccatt cgacccccat tatgtggaag tcctgggcga gcgcatgcac tacgtcgatg
     2101 ttggtccgcg cgatggcacc cctgtgctgt tcctgcacgg taacccgacc tcctcctacg
     2161 tgtggcgcaa catcatcccg catgttgcac cgacccatcg ctgcattgct ccagacctga
     2221 tcggtatggg caaatccgac aaaccagacc tgggttattt cttcgacgac cacgtccgct
     2281 tcatggatgc cttcatcgaa gccctgggtc tggaagaggt cgtcctggtc attcacgact
     2341 ggggctccgc tctgggtttc cactgggcca agcgcaatcc agagcgcgtc aaaggtattg
     2401 catttatgga gttcatccgc cctatcccga cctgggacga atggccagaa tttgcccgcg
     2461 agaccttcca ggccttccgc accaccgacg tcggccgcaa gctgatcatc gatcagaacg
     2521 tttttatcga gggtacgctg ccgatgggtg tcgtccgccc gctgactgaa gtcgagatgg
     2581 accattaccg cgagccgttc ctgaatcctg ttgaccgcga gccactgtgg cgcttcccaa
     2641 acgagctgcc aatcgccggt gagccagcga acatcgtcgc gctggtcgaa gaatacatgg
     2701 actggctgca ccagtcccct gtcccgaagc tgctgttctg gggcacccca ggcgttctga
     2761 tcccaccggc cgaagccgct cgcctggcca aaagcctgcc taactgcaag gctgtggaca
     2821 tcggcccggg tctgaatctg ctgcaagaag acaacccgga cctgatcggc agcgagatcg
     2881 cgcgctggct gtctactctg gagatttccg gtcatcatca ccatcaccac taatagtcaa
     2941 ccgctgagca ataactagca taaccccttg gggcctctaa acgggtcttg aggggttttt
     3001 tgctgaaagg aggaactata tccggttcgc ttgctgtcca taaaaccgcc cagtctagct
     3061 atcgccatgt aagcccactg caagctacct gctttctctt tgcgcttgcg ttttcccttg
     3121 tccagatagc ccagtagctg acattcatcc ggggtcagca ccgtttctgc ggactggctt
     3181 tctacgtaat ggtttcttag acgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc
     3241 ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct
     3301 gataaatgct tcaataatat tgaaaaagga agagtatgag tattcaacat ttccgtgtcg
     3361 cccttattcc cttttttgcg gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg
     3421 tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt gggttacatc gaactggatc
     3481 tcaacagcgg taagatcctt gagagttttc gccccgaaga acgttttcca atgatgagca
     3541 ctttcaaagt tctgctatgt ggcgcggtat tatcccgtat tgacgccggg caagagcaac
     3601 tcggtcgccg catacactat tctcagaatg acttggttga gtactcacca gtcacagaaa
     3661 agcatcttac ggatggcatg acagtaagag aattatgcag tgctgccata accatgagtg
     3721 ataacactgc ggccaactta cttctgacaa ctatcggagg accgaaggag ctaaccgctt
     3781 ttttgcacaa catgggggat catgtaactc gccttgatcg ttgggaaccg gagctgaatg
     3841 aagccatacc aaacgacgag cgtgacacca cgatgcctgt agcaatggca acaacgttgc
     3901 gcaaactatt aactggcgaa ctacttactc tagcttcccg gcaacaatta atagactgga
     3961 tggaggcgga taaagttgca ggaccacttc tgcgctcggc ccttccggct ggctggttta
     4021 ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc
     4081 cagatggtaa gccctcccgt atcgtagtta tctacacgac ggggagtcag gcaactatgg
     4141 atgaacgaaa tagacagatc gctgagatag gtgcctcact gattaagcat tggtaattcg
     4201 aaatgaccga ccaagcgacg cccaaccggt atcagctcac tcaaaggcgg taatacggtt
     4261 atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc
     4321 caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga
     4381 gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata
     4441 ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac
     4501 cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg
     4561 taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc
     4621 cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag
     4681 acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt
     4741 aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta gaaggacagt
     4801 atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg
     4861 atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac
     4921 gcgcagaaaa aaaggatttc aagaagatcc tttgatcttt tctacggggt ctgacgctca
     4981 gtggaacgaa aactcacgtt aagggcatat gatggtagtg tggggtctcc ccatgcgaga
     5041 gtagggaact gccaggcatc aaataaaacg aaaggctcag tcgaaagact gggcctttcg
     5101 ttttatctgt tgtttgtcgg tgaacgctct cctgagtagg acaaatccgc cgggagcgga
     5161 tttgaacgtt gcgaagcaac ggcccggagg gtggcgggca ggacgcccgc cat
//
