LOCUS       sequence-344839-        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-Halo
FEATURES             Location/Qualifiers
     source          1..5213
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      55..141
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      233..260
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     primer_bind     342..361
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        342..360
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      418..427
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2200..2220
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="EDLYFQS"
     CDS             2236..3120
                     /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             3121..3138
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(3149..3167)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      3163..3210
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        3439..3543
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3544..3612,3613..4404)
                     /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(3762..3781)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      complement(4562..5150)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4642..4661)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 catatgatgg tagtgtgggg tctccccatg cgagagtagg gaactgccag gcatcaaata
       61 aaacgaaagg ctcagtcgaa agactgggcc tttcgtttta tctgttgttt gtcggtgaac
      121 gctctcctga gtaggacaaa tccgccggga gcggatttga acgttgcgaa gcaacggccc
      181 ggagggtggc gggcaggacg cccgccataa actgccaggc atcaaattaa gcagaaggcc
      241 atcctgacgg atggcctttt tgcgtttcta caaactcttt tgtttatttt tctaaataca
      301 ttcaaatatg tatccgctca tggatctcga tcccgcgaaa ttaatacgac tcactatagg
      361 gtattttaac tttactaagg agcgatcgct tccgaattcc taccgcggat atctagagcc
      421 accatggcga acaagcttta catcggcaac ctcaacgaga gcgtgacccc cgcggacttg
      481 gagaaagtgt ttgcggagca caagatctcc tacagcggcc agttcttggt caaatccggc
      541 tacgccttcg tggactgccc ggacgagcac tgggcgatga aggccatcga aactttctcc
      601 gggaaagtag aattacaagg aaaacgctta gagattgaac attcggtgcc caaaaaacaa
      661 aggagccgga aaattcaaat ccgaaatatt ccaccccagc tccgatggga agtactggac
      721 agcctgctgg ctcagtatgg tacagtagag aactgtgagc aagtgaacac cgagagtgag
      781 acggcagtgg tgaatgtcac ctattccaac cgggagcaga ccaggcaagc catcatgaag
      841 ctgaatggcc accagttgga gaaccatgcc ctgaaggtct cctacatccc cgatgagcag
      901 atagcacagg gacctgagaa tgggcgccga gggggctttg gctctcgggg tcagccccgc
      961 cagggctcac ctgtggcagc gggggcccca gccaagcagc agcaagtgga catccccctt
     1021 cggctcctgg tgcccaccca gtatgtgggt gccattattg gcaaggaggg ggccaccatc
     1081 cgcaacatca caaaacagac ccagtccaag atagacgtgc ataggaagga gaacgcaggt
     1141 gcagctgaaa aagccatcag tgtgcactcc acccctgagg gctgctcctc cgcttgtaag
     1201 atgatcttgg agattatgca taaagaggct aaggacacca aaacggctga cgaggttccc
     1261 ctgaagatcc tggcccataa taactttgta gggcgtctca ttggcaagga aggacggaac
     1321 ctgaagaagg tagagcaaga taccgagaca aaaatcacca tctcctcgtt gcaagacctt
     1381 accctttaca accctgagag gaccatcact gtgaaggggg ccatcgagaa ttgttgcagg
     1441 gccgagcagg aaataatgaa gaaagttcgg gaggcctatg agaatgatgt ggctgccatg
     1501 agcctgcagt ctcacctgat ccctggcctg aacctggctg ctgtaggtct tttcccagct
     1561 tcatccagcg cagtcccgcc gcctcccagc agcgttactg gggctgctcc ctatagctcc
     1621 tttatgcagg ctcccgagca ggagatggtg caggtgttta tccccgccca ggcagtgggc
     1681 gccatcatcg gcaagaaggg gcagcacatc aaacagctct cccggtttgc cagcgcctcc
     1741 atcaagattg caccacccga aacacctgac tccaaagttc gtatggttat catcactgga
     1801 ccgccagagg cccaattcaa ggctcaggga agaatctatg gcaaactcaa ggaggagaac
     1861 ttctttggtc ccaaggagga agtgaagctg gagacccaca tacgtgtgcc agcatcagca
     1921 gctggccggg tcattggcaa aggtggaaaa acggtgaacg agttgcagaa tttgacggca
     1981 gctgaggtgg tagtaccaag agaccagacc cctgatgaga acgaccaggt catcgtgaaa
     2041 atcatcggac atttctatgc cagtcagatg gctcaacgga agatccgaga catcctggcc
     2101 caggttaagc agcagcatca gaagggacag agtaaccagg cccaggcacg gaggaagggc
     2161 ggctccggct cctgcaggcg agctctcgag ccaaccactg aggatctgta ctttcagagc
     2221 gataacgatg gatccgaaat cggtactggc tttccattcg acccccatta tgtggaagtc
     2281 ctgggcgagc gcatgcacta cgtcgatgtt ggtccgcgcg atggcacccc tgtgctgttc
     2341 ctgcacggta acccgacctc ctcctacgtg tggcgcaaca tcatcccgca tgttgcaccg
     2401 acccatcgct gcattgctcc agacctgatc ggtatgggca aatccgacaa accagacctg
     2461 ggttatttct tcgacgacca cgtccgcttc atggatgcct tcatcgaagc cctgggtctg
     2521 gaagaggtcg tcctggtcat tcacgactgg ggctccgctc tgggtttcca ctgggccaag
     2581 cgcaatccag agcgcgtcaa aggtattgca tttatggagt tcatccgccc tatcccgacc
     2641 tgggacgaat ggccagaatt tgcccgcgag accttccagg ccttccgcac caccgacgtc
     2701 ggccgcaagc tgatcatcga tcagaacgtt tttatcgagg gtacgctgcc gatgggtgtc
     2761 gtccgcccgc tgactgaagt cgagatggac cattaccgcg agccgttcct gaatcctgtt
     2821 gaccgcgagc cactgtggcg cttcccaaac gagctgccaa tcgccggtga gccagcgaac
     2881 atcgtcgcgc tggtcgaaga atacatggac tggctgcacc agtcccctgt cccgaagctg
     2941 ctgttctggg gcaccccagg cgttctgatc ccaccggccg aagccgctcg cctggccaaa
     3001 agcctgccta actgcaaggc tgtggacatc ggcccgggtc tgaatctgct gcaagaagac
     3061 aacccggacc tgatcggcag cgagatcgcg cgctggctgt ctactctgga gatttccggt
     3121 catcatcacc atcaccacta atagtcaacc gctgagcaat aactagcata accccttggg
     3181 gcctctaaac gggtcttgag gggttttttg ctgaaaggag gaactatatc cggttcgctt
     3241 gctgtccata aaaccgccca gtctagctat cgccatgtaa gcccactgca agctacctgc
     3301 tttctctttg cgcttgcgtt ttcccttgtc cagatagccc agtagctgac attcatccgg
     3361 ggtcagcacc gtttctgcgg actggctttc tacgtaatgg tttcttagac gtcaggtggc
     3421 acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat
     3481 atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag
     3541 agtatgagta ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt
     3601 cctgtttttg ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt
     3661 gcacgagtgg gttacatcga actggatctc aacagcggta agatccttga gagttttcgc
     3721 cccgaagaac gttttccaat gatgagcact ttcaaagttc tgctatgtgg cgcggtatta
     3781 tcccgtattg acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac
     3841 ttggttgagt actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa
     3901 ttatgcagtg ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaact
     3961 atcggaggac cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc
     4021 cttgatcgtt gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg
     4081 atgcctgtag caatggcaac aacgttgcgc aaactattaa ctggcgaact acttactcta
     4141 gcttcccggc aacaattaat agactggatg gaggcggata aagttgcagg accacttctg
     4201 cgctcggccc ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggg
     4261 tctcgcggta tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc
     4321 tacacgacgg ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt
     4381 gcctcactga ttaagcattg gtaattcgaa atgaccgacc aagcgacgcc caaccggtat
     4441 cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga
     4501 acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt
     4561 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt
     4621 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc
     4681 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa
     4741 gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct
     4801 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta
     4861 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg
     4921 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc
     4981 ctaactacgg ctacactaga aggacagtat ttggtatctg cgctctgctg aagccagtta
     5041 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg
     5101 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatttcaa gaagatcctt
     5161 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa ggg
//
