LOCUS       sequence-345604-        3984 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 3984)
  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 3984)
  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_MANF-Halo
FEATURES             Location/Qualifiers
     source          1..3984
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      5..32
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     primer_bind     114..133
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        114..132
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             743..763
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="EDLYFQS"
     CDS             779..1663
                     /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             1664..1681
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(1692..1710)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1706..1753
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        1982..2086
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2087..2155,2156..2947)
                     /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(2305..2324)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      complement(3105..3693)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3185..3204)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      3811..3897
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
ORIGIN
        1 aagcagaagg ccatcctgac ggatggcctt tttgcgtttc tacaaactct tttgtttatt
       61 tttctaaata cattcaaata tgtatccgct catggatctc gatcccgcga aattaatacg
      121 actcactata gggtatttta actttactaa ggagcgatcg ctttcgaatt cctaccgcgg
      181 atatatgagg aggatgtggg ccacgcaggg gctggcggtg gcgctggctc tgagcgtgct
      241 gccgggcagc cgggcgctgc ggccgggcga ctgcgaagtt tgtatttctt atctgggaag
      301 attttaccag gacctcaaag acagagatgt cacattctca ccagccacta ttgaaaacga
      361 acttataaag ttctgccggg aagcaagagg caaagagaat cggttgtgct actatatcgg
      421 ggccacagat gatgcagcca ccaaaatcat caatgaggta tcaaagcctc tggcccacca
      481 catccctgtg gagaagatct gtgagaagct taagaagaag gacagccaga tatgtgagct
      541 taagtatgac aagcagatcg acctgagcac agtggacctg aagaagctcc gagttaaaga
      601 gctgaagaag attctggatg actgggggga gacatgcaaa ggctgtgcag aaaagtctga
      661 ctacatccgg aagataaatg aactgatgcc taaatatgcc cccaaggcag ccagtgcacg
      721 gaccgatttg gggccaacca ctgaggatct gtactttcag agcgataacg atggatccga
      781 aatcggtact ggctttccat tcgaccccca ttatgtggaa gtcctgggcg agcgcatgca
      841 ctacgtcgat gttggtccgc gcgatggcac ccctgtgctg ttcctgcacg gtaacccgac
      901 ctcctcctac gtgtggcgca acatcatccc gcatgttgca ccgacccatc gctgcattgc
      961 tccagacctg atcggtatgg gcaaatccga caaaccagac ctgggttatt tcttcgacga
     1021 ccacgtccgc ttcatggatg ccttcatcga agccctgggt ctggaagagg tcgtcctggt
     1081 cattcacgac tggggctccg ctctgggttt ccactgggcc aagcgcaatc cagagcgcgt
     1141 caaaggtatt gcatttatgg agttcatccg ccctatcccg acctgggacg aatggccaga
     1201 atttgcccgc gagaccttcc aggccttccg caccaccgac gtcggccgca agctgatcat
     1261 cgatcagaac gtttttatcg agggtacgct gccgatgggt gtcgtccgcc cgctgactga
     1321 agtcgagatg gaccattacc gcgagccgtt cctgaatcct gttgaccgcg agccactgtg
     1381 gcgcttccca aacgagctgc caatcgccgg tgagccagcg aacatcgtcg cgctggtcga
     1441 agaatacatg gactggctgc accagtcccc tgtcccgaag ctgctgttct ggggcacccc
     1501 aggcgttctg atcccaccgg ccgaagccgc tcgcctggcc aaaagcctgc ctaactgcaa
     1561 ggctgtggac atcggcccgg gtctgaatct gctgcaagaa gacaacccgg acctgatcgg
     1621 cagcgagatc gcgcgctggc tgtctactct ggagatttcc ggtcatcatc accatcacca
     1681 ctaatagtca accgctgagc aataactagc ataacccctt ggggcctcta aacgggtctt
     1741 gaggggtttt ttgctgaaag gaggaactat atccggttcg cttgctgtcc ataaaaccgc
     1801 ccagtctagc tatcgccatg taagcccact gcaagctacc tgctttctct ttgcgcttgc
     1861 gttttccctt gtccagatag cccagtagct gacattcatc cggggtcagc accgtttctg
     1921 cggactggct ttctacgtaa tggtttctta gacgtcaggt ggcacttttc ggggaaatgt
     1981 gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag
     2041 acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtatga gtattcaaca
     2101 tttccgtgtc gcccttattc ccttttttgc ggcattttgc cttcctgttt ttgctcaccc
     2161 agaaacgctg gtgaaagtaa aagatgctga agatcagttg ggtgcacgag tgggttacat
     2221 cgaactggat ctcaacagcg gtaagatcct tgagagtttt cgccccgaag aacgttttcc
     2281 aatgatgagc actttcaaag ttctgctatg tggcgcggta ttatcccgta ttgacgccgg
     2341 gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat gacttggttg agtactcacc
     2401 agtcacagaa aagcatctta cggatggcat gacagtaaga gaattatgca gtgctgccat
     2461 aaccatgagt gataacactg cggccaactt acttctgaca actatcggag gaccgaagga
     2521 gctaaccgct tttttgcaca acatggggga tcatgtaact cgccttgatc gttgggaacc
     2581 ggagctgaat gaagccatac caaacgacga gcgtgacacc acgatgcctg tagcaatggc
     2641 aacaacgttg cgcaaactat taactggcga actacttact ctagcttccc ggcaacaatt
     2701 aatagactgg atggaggcgg ataaagttgc aggaccactt ctgcgctcgg cccttccggc
     2761 tggctggttt attgctgata aatctggagc cggtgagcgt gggtctcgcg gtatcattgc
     2821 agcactgggg ccagatggta agccctcccg tatcgtagtt atctacacga cggggagtca
     2881 ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac tgattaagca
     2941 ttggtaattc gaaatgaccg accaagcgac gcccaaccgg tatcagctca ctcaaaggcg
     3001 gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc
     3061 cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc
     3121 ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga
     3181 ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc
     3241 ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat
     3301 agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg
     3361 cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc
     3421 aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga
     3481 gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact
     3541 agaaggacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt
     3601 ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag
     3661 cagcagatta cgcgcagaaa aaaaggattt caagaagatc ctttgatctt ttctacgggg
     3721 tctgacgctc agtggaacga aaactcacgt taagggcata tgatggtagt gtggggtctc
     3781 cccatgcgag agtagggaac tgccaggcat caaataaaac gaaaggctca gtcgaaagac
     3841 tgggcctttc gttttatctg ttgtttgtcg gtgaacgctc tcctgagtag gacaaatccg
     3901 ccgggagcgg atttgaacgt tgcgaagcaa cggcccggag ggtggcgggc aggacgcccg
     3961 ccataaactg ccaggcatca aatt
//
