LOCUS       sequence-274443-        7362 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses the hyperactive form of the catalytic domain of the ADAR 
            protein fused to HRP-48 for use in ""TRIBE"" assays.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7362)
  AUTHORS   Xu W, Rahman R, Rosbash M
  TITLE     Mechanistic implications of enhanced editing by a HyperTRIBE 
            RNA-binding protein.
  JOURNAL   RNA. 2018 Feb;24(2):173-182. doi: 10.1261/rna.064691.117. Epub 2017 
            Nov 10.
  PUBMED    29127211
REFERENCE   2  (bases 1 to 7362)
  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"; doi: 
            "10.1261/rna.064691.117"; journalName: "RNA"; date: "2018-02"; 
            volume: "24"; issue: "2"; pages: "173-182"
COMMENT     Sequence Label: pMT-Hrp48-ADARcd-E488Q-V5
FEATURES             Location/Qualifiers
     source          1..7362
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(29..51)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     151..170
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     364..386
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     378..395
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     379..395
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        401..827
                     /label=MT promoter
                     /note="Drosophila metallothionein promoter"
     CDS             3520..3561
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             3571..3588
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(3611..3628)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3630..3764
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3908..3924)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3908..3924)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3921..3943)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3932..3948
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(3956..3962,3963..3980,3981..3986))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4001..4022
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        4207..4486
                     /label=copia promoter
                     /note="strong promoter from the Drosophila transposable 
                     element copia (Sinclair et al., 1986)"
     CDS             4503..4901
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     polyA_signal    5230..5364
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5372..5389)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5543..6131)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5623..5642)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6302..7093,7094..7162))
                     /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     6925..6944
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7163..7267)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca
       61 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg
      121 ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc
      181 accatatgcg gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc atcaggcgcc
      241 attcgccatt caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat
      301 tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta acgccagggt
      361 tttcccagtc acgacgttgt aaaacgacgg ccagtgaatt aattcgttgc aggacaggat
      421 gtggtgcccg atgtgactag ctctttgctg caggccgtcc tatcctctgg ttccgataag
      481 agacccagaa ctccggcccc ccaccgccca ccgccacccc catacatatg tggtacgcaa
      541 gtaagagtgc ctgcgcatgc cccatgtgcc ccaccaagag ctttgcatcc catacaagtc
      601 cccaaagtgg agaaccgaac caattcttcg cgggcagaac aaaagcttct gcacacgtct
      661 ccactcgaat ttggagccgg ccggcgtgtg caaaagaggt gaatcgaacg aaagacccgt
      721 gtgtaaagcc gcgtttccaa aatgtataaa accgagagca tctggccaat gtgcatcagt
      781 tgtggtcagc agcaaaatca agtgaatcat ctcagtgcaa ctaaaggggg gatctagatc
      841 ggggtacctc gaaggccgcc atatgctagc aatggaggaa gacgagaggg gcaaactttt
      901 tgtgggcggt ctatcctggg agacgacgca ggagaacctg tcgcgctact tctgccgctt
      961 cggggacatc attgactgtg tggtgatgaa gaacaacgag agcggcaggt cgcgcggctt
     1021 tggcttcgtt accttcgccg atcccaccaa cgtcaaccac gtgctgcaga acggaccgca
     1081 cacgctcgac ggtcgcacca tcgaccccaa gccgtgcaat ccgcgcactc tgcagaaacc
     1141 gaagaagggc ggcggctaca aggtcttcct gggtggcctg ccctcgaacg tcaccgagac
     1201 cgatctgcgg accttcttta accgctacgg caaggtcacc gaggtggtta tcatgtacga
     1261 ccaggagaag aagaagtccc gcggcttcgg ctttctctcc ttcgaggagg agtcctccgt
     1321 tgagcacgtg accaacgagc ggtacatcaa tctgaatggc aagcaggtcg aaatcaagaa
     1381 ggccgagcct cgtgatggat ctggcggcca aaactccaac aacagtaccg tgggaggcgc
     1441 ctatggcaag cttggtaacg agtgcagcca ctggggaccg caccatgctc ccatcaacat
     1501 gatgcagggc cagaatggcc agatgggtgg accgccgctg aatatgccca ttggagcgcc
     1561 gaatatgatg cctggctatc agggttgggg cacctcgccg cagcagcaac aatacggcta
     1621 cggcaacagt ggcccaggat cgtaccaggg atggggagct ccaccaggac cccagggacc
     1681 accaccgcag tggtcgaact acgctggacc tcagcagacg cagggctacg gcggatacga
     1741 catgtataac tcgacgtcga ccggagctcc ttcgggacca tcgggcggcg gcagctggaa
     1801 ctcgtggaac atgccaccta actctgccgg acccactggg gcaccaggag ccggagcggg
     1861 caccgccact gacatgtact cgcgtgctca agcctgggcg acgggcggtc cctcgaccac
     1921 tggaccagtg ggcggcatgc cccggaccgg acccggtaat tcggcctcca agtctggctc
     1981 tgagtacgac tacggcggct atggatccgg gtacgactac gactacagca actatgtgaa
     2041 gcaggagggc gcctcgaact acggagcagg gccgcgatca gcgtacggca acgacagctc
     2101 cacgcagcca ccctatgcaa cctcgcaggc tgtagcggcc gccaccgcgg tggactacag
     2161 tccaatggtg gtgccacaga agaacgtacc cctgccaatc gacgacaagt cgtcatcgat
     2221 ggagttgcct cagatacacg cggatacgat tggtcggttg gtcttagaaa agttcatgga
     2281 agtaatcaag ggccaggagg cttactcgcg tcgaaaggta ttagcgggca ttgtaatgac
     2341 tgaaaacatg aatttttgtg aagccaaagt tatttcagtt tcgacgggca ccaagtgtgt
     2401 cagcggtgag catatgagtg tgaacggagc tgtcctaaat gattcccatg ctgaaatagt
     2461 ctccaggcgt tgtctcctca aatatttata tgcacagctg gaccttcagt gcaatcaggc
     2521 cacagcatat cagtcgattt tcgtgaggaa tactgatggg caataccctt ataaactaaa
     2581 atccggggta catttccatt tgtatataaa tacagcacct tgtggggatg cacggatatt
     2641 tagtcctcac gaaaacgaca ctggtgttga taaacatcca aatagaaaag ctcgaggcca
     2701 gctgcgcacc aaaatcgagt ccggtcaggg gacgattcca gtgaaaagca gtgatggtat
     2761 ccagacgtgg gatggcgtat tgcaggtaac ccgattgggc ggttgctatt agatcgtggt
     2821 aaaccatcaa cattacaaca tttatgactt ttgtgcaggg ccaacgcctg ctaacaatgt
     2881 cgtgctcgga taaaattgcc cgttggaaca tcgtgggcat ccaaggctca ctattgtctt
     2941 ccataattga accggtgtac ctgcattcga ttgtgctggg aagcctgttg catccggagc
     3001 acatgtaccg cgcagtttgc ggccgaattg agaagtccat tcaaggcctg ccaccgccgt
     3061 accatctgaa caagccgcgc ctggcactag tcacttcggc cgagccgcgg aatcaggcca
     3121 aggctcccaa cttcgggatc aattggacca ttggggacac cgagctggaa gtggtgaact
     3181 cgctaaccgg tcgaaccatt ggcggccagg tgtctcgcat cacaaagcag gcgtttttcg
     3241 ttaagtatgg atttctaatg gcaaacttgc ccggtatttt agtccgcaaa gtaaccactg
     3301 actatgggca aaccaaagct aacgttaagg actatcaggt gagtgtttat atgctggatc
     3361 atttgaggat atgcatcaaa ttgtttgcca catctgttct tgtagatcgc aaagctagaa
     3421 ttgttctctg cattcaagcg agaagacctt ggcagctggc tgaagaaacc cattgaacaa
     3481 gacgagttcg gtcttgccga atctagaggg cccttcgaag gtaagcctat ccctaaccct
     3541 ctcctcggtc tcgattctac gcgtaccggt catcatcacc atcaccattg agtttaaacc
     3601 cgctgatcag cctcgactgt gccttctaag atccagacat gataagatac attgatgagt
     3661 ttggacaaac cacaactaga atgcagtgaa aaaaatgctt tatttgtgaa atttgtgatg
     3721 ctattgcttt atttgtaacc attataagct gcaataaaca agttaacaac aacaattgca
     3781 ttcattttat gtttcaggtt cagggggagg tgtgggaggt tttttaaagc aagtaaaacc
     3841 tctacaaatg tggtatggct gattatgatc agtcgacctg caggcatgca agcttggcgt
     3901 aatcatggtc atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca
     3961 tacgagccgg aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat
     4021 taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt
     4081 aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgctatcc
     4141 ggtgcgtggt ggtttcatgc ttctgggaac ggcaaatggg tttaggattg ggaacccctc
     4201 atcatctgtt ggaatatact attcaaccta caaaagtaac gttaaacaac actactttat
     4261 atttgatatg aatggccaca ccttttatgc cataaaacat attgtaagag aataccactc
     4321 tttttattcc ttctttcctt cttgtacgtt ttttgctgta agtaggtcgt ggtgctggtg
     4381 ttgcagttga aataacttaa aatataaatc ataaaactca aacataaact tgactattta
     4441 tttatttatt aagaaaggaa atataaattg taaattacaa caggttccct ttatgcgaag
     4501 ggatggccaa gcctttgtct caagaagaat ccaccctcat tgaaagagca acggctacaa
     4561 tcaacagcat ccccatctct gaagactaca gcgtcgccag cgcagctctc tctagcgacg
     4621 gccgcatctt cactggtgtc aatgtatatc attttactgg gggaccttgc gcagaactcg
     4681 tggtgctggg cactgctgct gctgcggcag ctggcaacct gacttgtatc gtcgcgatcg
     4741 gaaatgagaa caggggcatc ttgagcccct gcggacggtg ccgacaggtt cttctcgatc
     4801 tgcatcctgg gatcaaagcc atagtgaagg acagtgatgg acagccgacg gcagttggga
     4861 ttcgtgaatt gctgccctct ggttatgtgt gggagggcta acccttatac gcaagggagt
     4921 agatgccgac cgaacaagag ctgatttcga gaacgcctca gccagcaact cgcgcgagcc
     4981 tagcaaggca aatgcgagag aacggcctta cgcttggtgg cacagttctc gtccacagtt
     5041 cgctaagctc gctcggctgg gtcgcgggag ggccggtcgc agtgattcag gcccttctgg
     5101 attgtgttgg tccccagggc acgattgtca tgcccacgca ctcgggtgat ctgactgatc
     5161 ccgcagattg gagatcgccg cccgtgcctg ccgattgggt gcagatcagc ctcgaggcca
     5221 gctagcttga acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca
     5281 aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc
     5341 aatgtatctt atcatgtctg gatcgggctt cctcgctcac tgactcgctg cgctcggtcg
     5401 ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat
     5461 caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta
     5521 aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa
     5581 atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc
     5641 cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt
     5701 ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca
     5761 gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg
     5821 accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat
     5881 cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta
     5941 cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct
     6001 gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac
     6061 aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa
     6121 aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa
     6181 actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt
     6241 taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca
     6301 gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca
     6361 tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc
     6421 ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa
     6481 accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc
     6541 agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca
     6601 acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat
     6661 tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag
     6721 cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac
     6781 tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt
     6841 ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt
     6901 gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc
     6961 tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat
     7021 ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca
     7081 gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga
     7141 cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg
     7201 gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg
     7261 ttccgcgcac atttccccga aaagtgccac ctgacgtcta agaaaccatt attatcatga
     7321 cattaaccta taaaaatagg cgtatcacga ggccctttcg tc
//
