LOCUS       sequence-345072-        5235 bp    DNA     circular SYN 14-APR-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 Apr 14, 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_IGF2BP2a-Halo
FEATURES             Location/Qualifiers
     source          1..5235
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      78..164
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      256..283
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     primer_bind     365..384
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        365..383
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             2245..2265
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="EDLYFQS"
     CDS             2281..3165
                     /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             3166..3183
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(3194..3212)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      3208..3255
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        3484..3588
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3589..3657,3658..4449)
                     /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(3807..3826)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      complement(4607..5195)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4687..4706)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ggaacgaaaa ctcacgttaa gggcatatga tggtagtgtg gggtctcccc atgcgagagt
       61 agggaactgc caggcatcaa ataaaacgaa aggctcagtc gaaagactgg gcctttcgtt
      121 ttatctgttg tttgtcggtg aacgctctcc tgagtaggac aaatccgccg ggagcggatt
      181 tgaacgttgc gaagcaacgg cccggagggt ggcgggcagg acgcccgcca taaactgcca
      241 ggcatcaaat taagcagaag gccatcctga cggatggcct ttttgcgttt ctacaaactc
      301 ttttgtttat ttttctaaat acattcaaat atgtatccgc tcatggatct cgatcccgcg
      361 aaattaatac gactcactat agggtatttt aactttacta aggagcgatc gcttccgaat
      421 tcctaccgcg gatatatgat gaacaagctt tacatcggga acctgagccc cgccgtcacc
      481 gccgacgacc tccggcagct ctttggggac aggaagctgc ccctggcggg acaggtcctg
      541 ctgaagtccg gctacgcctt cgtggactac cccgaccaga actgggccat ccgcgccatc
      601 gagaccctct cgggtaaagt ggaattgcat gggaaaatca tggaagttga ttactcagtc
      661 tctaaaaagc taaggagcag gaaaattcag attcgaaaca tccctcctca cctgcagtgg
      721 gaggtgttgg atggactttt ggctcaatat gggacagtgg agaatgtgga acaagtcaac
      781 acagacacag aaaccgccgt tgtcaacgtc acatatgcaa caagagaaga agcaaaaata
      841 gccatggaga agctaagcgg gcatcagttt gagaactact ccttcaagat ttcctacatc
      901 ccggatgaag aggtgagctc cccttcgccc cctcagcgag cccagcgtgg ggaccactct
      961 tcccgggagc aaggccacgc ccctgggggc acttctcagg ccagacagat tgatttcccg
     1021 ctgcggatcc tggtccccac ccagtttgtt ggtgccatca tcggaaagga gggcttgacc
     1081 ataaagaaca tcactaagca gacccagtcc cgggtagata tccatagaaa agagaactct
     1141 ggagctgcag agaagcctgt caccatccat gccaccccag aggggacttc tgaagcatgc
     1201 cgcatgattc ttgaaatcat gcagaaagag gcagatgaga ccaaactagc cgaagagatt
     1261 cctctgaaaa tcttggcaca caatggcttg gttggaagac tgattggaaa agaaggcaga
     1321 aatttgaaga aaattgaaca tgaaacaggg accaagataa caatctcatc tttgcaggat
     1381 ttgagcatat acaacccgga aagaaccatc actgtgaagg gcacagttga ggcctgtgcc
     1441 agtgctgaga tagagattat gaagaagctg cgtgaggcct ttgaaaatga tatgctggct
     1501 gttaaccaac aagccaatct gatcccaggg ttgaacctca gcgcacttgg catcttttca
     1561 acaggactgt ccgtgctatc tccaccagca gggccccgcg gagctccccc cgctgccccc
     1621 taccacccct tcactaccca ctccggatac ttctccagcc tgtaccccca tcaccagttt
     1681 ggcccgttcc cgcatcatca ctcttatcca gagcaggaga ttgtgaatct cttcatccca
     1741 acccaggctg tgggcgccat catcgggaag aagggggcac acatcaaaca gctggcgaga
     1801 ttcgccggag cctctatcaa gattgcccct gcggaaggcc cagacgtcag cgaaaggatg
     1861 gtcatcatca ccgggccacc ggaagcccag ttcaaggccc agggacggat ctttgggaaa
     1921 ctgaaagagg aaaacttctt taaccccaaa gaagaagtga agctggaagc gcatatcaga
     1981 gtgccctctt ccacagctgg ccgggtgatt ggcaaaggtg gcaagaccgt gaacgaactg
     2041 cagaacttaa ccagtgcaga agtcatcgtg cctcgtgacc aaacgccaga tgaaaatgag
     2101 gaagtgatcg tcagaattat cgggcacttc tttgctagcc agactgcaca gcgcaagatc
     2161 agggaaattg tacaacaggt gaagcagcag gagcagaaat accctcaggg agtcgcctca
     2221 cagcgcagca aggggccaac cactgaggat ctgtactttc agagcgataa cgatggatcc
     2281 gaaatcggta ctggctttcc attcgacccc cattatgtgg aagtcctggg cgagcgcatg
     2341 cactacgtcg atgttggtcc gcgcgatggc acccctgtgc tgttcctgca cggtaacccg
     2401 acctcctcct acgtgtggcg caacatcatc ccgcatgttg caccgaccca tcgctgcatt
     2461 gctccagacc tgatcggtat gggcaaatcc gacaaaccag acctgggtta tttcttcgac
     2521 gaccacgtcc gcttcatgga tgccttcatc gaagccctgg gtctggaaga ggtcgtcctg
     2581 gtcattcacg actggggctc cgctctgggt ttccactggg ccaagcgcaa tccagagcgc
     2641 gtcaaaggta ttgcatttat ggagttcatc cgccctatcc cgacctggga cgaatggcca
     2701 gaatttgccc gcgagacctt ccaggccttc cgcaccaccg acgtcggccg caagctgatc
     2761 atcgatcaga acgtttttat cgagggtacg ctgccgatgg gtgtcgtccg cccgctgact
     2821 gaagtcgaga tggaccatta ccgcgagccg ttcctgaatc ctgttgaccg cgagccactg
     2881 tggcgcttcc caaacgagct gccaatcgcc ggtgagccag cgaacatcgt cgcgctggtc
     2941 gaagaataca tggactggct gcaccagtcc cctgtcccga agctgctgtt ctggggcacc
     3001 ccaggcgttc tgatcccacc ggccgaagcc gctcgcctgg ccaaaagcct gcctaactgc
     3061 aaggctgtgg acatcggccc gggtctgaat ctgctgcaag aagacaaccc ggacctgatc
     3121 ggcagcgaga tcgcgcgctg gctgtctact ctggagattt ccggtcatca tcaccatcac
     3181 cactaatagt caaccgctga gcaataacta gcataacccc ttggggcctc taaacgggtc
     3241 ttgaggggtt ttttgctgaa aggaggaact atatccggtt cgcttgctgt ccataaaacc
     3301 gcccagtcta gctatcgcca tgtaagccca ctgcaagcta cctgctttct ctttgcgctt
     3361 gcgttttccc ttgtccagat agcccagtag ctgacattca tccggggtca gcaccgtttc
     3421 tgcggactgg ctttctacgt aatggtttct tagacgtcag gtggcacttt tcggggaaat
     3481 gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg
     3541 agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtat gagtattcaa
     3601 catttccgtg tcgcccttat tccctttttt gcggcatttt gccttcctgt ttttgctcac
     3661 ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg agtgggttac
     3721 atcgaactgg atctcaacag cggtaagatc cttgagagtt ttcgccccga agaacgtttt
     3781 ccaatgatga gcactttcaa agttctgcta tgtggcgcgg tattatcccg tattgacgcc
     3841 gggcaagagc aactcggtcg ccgcatacac tattctcaga atgacttggt tgagtactca
     3901 ccagtcacag aaaagcatct tacggatggc atgacagtaa gagaattatg cagtgctgcc
     3961 ataaccatga gtgataacac tgcggccaac ttacttctga caactatcgg aggaccgaag
     4021 gagctaaccg cttttttgca caacatgggg gatcatgtaa ctcgccttga tcgttgggaa
     4081 ccggagctga atgaagccat accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg
     4141 gcaacaacgt tgcgcaaact attaactggc gaactactta ctctagcttc ccggcaacaa
     4201 ttaatagact ggatggaggc ggataaagtt gcaggaccac ttctgcgctc ggcccttccg
     4261 gctggctggt ttattgctga taaatctgga gccggtgagc gtgggtctcg cggtatcatt
     4321 gcagcactgg ggccagatgg taagccctcc cgtatcgtag ttatctacac gacggggagt
     4381 caggcaacta tggatgaacg aaatagacag atcgctgaga taggtgcctc actgattaag
     4441 cattggtaat tcgaaatgac cgaccaagcg acgcccaacc ggtatcagct cactcaaagg
     4501 cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag
     4561 gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc
     4621 gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag
     4681 gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga
     4741 ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc
     4801 atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg
     4861 tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt
     4921 ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca
     4981 gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca
     5041 ctagaaggac agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag
     5101 ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca
     5161 agcagcagat tacgcgcaga aaaaaaggat ttcaagaaga tcctttgatc ttttctacgg
     5221 ggtctgacgc tcagt
//
