LOCUS       sequence-194903-        7362 bp    DNA     circular SYN 14-APR-2025
DEFINITION  identification of cell-specific targets of RNA binding proteins.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7362)
  AUTHORS   McMahon AC, Rahman R, Jin H, Shen JL, Fieldsend A, Luo W, Rosbash M
  TITLE     TRIBE: Hijacking an RNA-Editing Enzyme to Identify Cell-Specific 
            Targets of RNA-Binding Proteins.
  JOURNAL   Cell. 2016 Mar 30. pii: S0092-8674(16)30263-X. doi: 
            10.1016/j.cell.2016.03.007.
  PUBMED    27040499
REFERENCE   2  (bases 1 to 7362)
  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: "Cell. 2016 
            Mar 30. pii: S0092-8674(16)30263-X. doi: 
            10.1016/j.cell.2016.03.007."
COMMENT     Sequence Label: pMT-Hrp48-ADARcd-V5
FEATURES             Location/Qualifiers
     source          1..7362
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..427
                     /label=MT promoter
                     /note="Drosophila metallothionein promoter"
     CDS             3120..3161
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             3171..3188
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(3211..3228)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3230..3364
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3508..3524)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3508..3524)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3521..3543)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3532..3548
                     /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(3556..3562,3563..3580,3581..3586))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3601..3622
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        3807..4086
                     /label=copia promoter
                     /note="strong promoter from the Drosophila transposable 
                     element copia (Sinclair et al., 1986)"
     CDS             4103..4501
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     polyA_signal    4830..4964
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4972..4989)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5143..5731)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5223..5242)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5902..6693,6694..6762))
                     /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     6525..6544
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6763..6867)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(6991..7013)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     7113..7132
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     7326..7348
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     7340..7357
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     7341..7357
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 aattcgttgc aggacaggat gtggtgcccg atgtgactag ctctttgctg caggccgtcc
       61 tatcctctgg ttccgataag agacccagaa ctccggcccc ccaccgccca ccgccacccc
      121 catacatatg tggtacgcaa gtaagagtgc ctgcgcatgc cccatgtgcc ccaccaagag
      181 ctttgcatcc catacaagtc cccaaagtgg agaaccgaac caattcttcg cgggcagaac
      241 aaaagcttct gcacacgtct ccactcgaat ttggagccgg ccggcgtgtg caaaagaggt
      301 gaatcgaacg aaagacccgt gtgtaaagcc gcgtttccaa aatgtataaa accgagagca
      361 tctggccaat gtgcatcagt tgtggtcagc agcaaaatca agtgaatcat ctcagtgcaa
      421 ctaaaggggg gatctagatc ggggtacctc gaaggccgcc atatgctagc aatggaggaa
      481 gacgagaggg gcaaactttt tgtgggcggt ctatcctggg agacgacgca ggagaacctg
      541 tcgcgctact tctgccgctt cggggacatc attgactgtg tggtgatgaa gaacaacgag
      601 agcggcaggt cgcgcggctt tggcttcgtt accttcgccg atcccaccaa cgtcaaccac
      661 gtgctgcaga acggaccgca cacgctcgac ggtcgcacca tcgaccccaa gccgtgcaat
      721 ccgcgcactc tgcagaaacc gaagaagggc ggcggctaca aggtcttcct gggtggcctg
      781 ccctcgaacg tcaccgagac cgatctgcgg accttcttta accgctacgg caaggtcacc
      841 gaggtggtta tcatgtacga ccaggagaag aagaagtccc gcggcttcgg ctttctctcc
      901 ttcgaggagg agtcctccgt tgagcacgtg accaacgagc ggtacatcaa tctgaatggc
      961 aagcaggtcg aaatcaagaa ggccgagcct cgtgatggat ctggcggcca aaactccaac
     1021 aacagtaccg tgggaggcgc ctatggcaag cttggtaacg agtgcagcca ctggggaccg
     1081 caccatgctc ccatcaacat gatgcagggc cagaatggcc agatgggtgg accgccgctg
     1141 aatatgccca ttggagcgcc gaatatgatg cctggctatc agggttgggg cacctcgccg
     1201 cagcagcaac aatacggcta cggcaacagt ggcccaggat cgtaccaggg atggggagct
     1261 ccaccaggac cccagggacc accaccgcag tggtcgaact acgctggacc tcagcagacg
     1321 cagggctacg gcggatacga catgtataac tcgacgtcga ccggagctcc ttcgggacca
     1381 tcgggcggcg gcagctggaa ctcgtggaac atgccaccta actctgccgg acccactggg
     1441 gcaccaggag ccggagcggg caccgccact gacatgtact cgcgtgctca agcctgggcg
     1501 acgggcggtc cctcgaccac tggaccagtg ggcggcatgc cccggaccgg acccggtaat
     1561 tcggcctcca agtctggctc tgagtacgac tacggcggct atggatccgg gtacgactac
     1621 gactacagca actatgtgaa gcaggagggc gcctcgaact acggagcagg gccgcgatca
     1681 gcgtacggca acgacagctc cacgcagcca ccctatgcaa cctcgcaggc tgtagcggcc
     1741 gccaccgcgg tggactacag tccaatggtg gtgccacaga agaacgtacc cctgccaatc
     1801 gacgacaagt cgtcatcgat ggagttgcct cagatacacg cggatacgat tggtcggttg
     1861 gtcttagaaa agttcatgga agtaatcaag ggccaggagg cttactcgcg tcgaaaggta
     1921 ttagcgggca ttgtaatgac tgaaaacatg aatttttgtg aagccaaagt tatttcagtt
     1981 tcgacgggca ccaagtgtgt cagcggtgag catatgagtg tgaacggagc tgtcctaaat
     2041 gattcccatg ctgaaatagt ctccaggcgt tgtctcctca aatatttata tgcacagctg
     2101 gaccttcagt gcaatcaggc cacagcatat cagtcgattt tcgtgaggaa tactgatggg
     2161 caataccctt ataaactaaa atccggggta catttccatt tgtatataaa tacagcacct
     2221 tgtggggatg cacggatatt tagtcctcac gaaaacgaca ctggtgttga taaacatcca
     2281 aatagaaaag ctcgaggcca gctgcgcacc aaaatcgagt ccggtgaggg gacgattcca
     2341 gtgaaaagca gtgatggtat ccagacgtgg gatggcgtat tgcaggtaac ccgattgggc
     2401 ggttgctatt agatcgtggt aaaccatcaa cattacaaca tttatgactt ttgtgcaggg
     2461 ccaacgcctg ctaacaatgt cgtgctcgga taaaattgcc cgttggaaca tcgtgggcat
     2521 ccaaggctca ctattgtctt ccataattga accggtgtac ctgcattcga ttgtgctggg
     2581 aagcctgttg catccggagc acatgtaccg cgcagtttgc ggccgaattg agaagtccat
     2641 tcaaggcctg ccaccgccgt accatctgaa caagccgcgc ctggcactag tcacttcggc
     2701 cgagccgcgg aatcaggcca aggctcccaa cttcgggatc aattggacca ttggggacac
     2761 cgagctggaa gtggtgaact cgctaaccgg tcgaaccatt ggcggccagg tgtctcgcat
     2821 cacaaagcag gcgtttttcg ttaagtatgg atttctaatg gcaaacttgc ccggtatttt
     2881 agtccgcaaa gtaaccactg actatgggca aaccaaagct aacgttaagg actatcaggt
     2941 gagtgtttat atgctggatc atttgaggat atgcatcaaa ttgtttgcca catctgttct
     3001 tgtagatcgc aaagctagaa ttgttctctg cattcaagcg agaagacctt ggcagctggc
     3061 tgaagaaacc cattgaacaa gacgagttcg gtcttgccga atctagaggg cccttcgaag
     3121 gtaagcctat ccctaaccct ctcctcggtc tcgattctac gcgtaccggt catcatcacc
     3181 atcaccattg agtttaaacc cgctgatcag cctcgactgt gccttctaag atccagacat
     3241 gataagatac attgatgagt ttggacaaac cacaactaga atgcagtgaa aaaaatgctt
     3301 tatttgtgaa atttgtgatg ctattgcttt atttgtaacc attataagct gcaataaaca
     3361 agttaacaac aacaattgca ttcattttat gtttcaggtt cagggggagg tgtgggaggt
     3421 tttttaaagc aagtaaaacc tctacaaatg tggtatggct gattatgatc agtcgacctg
     3481 caggcatgca agcttggcgt aatcatggtc atagctgttt cctgtgtgaa attgttatcc
     3541 gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta
     3601 atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa
     3661 cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat
     3721 tgggcgctct tccgctatcc ggtgcgtggt ggtttcatgc ttctgggaac ggcaaatggg
     3781 tttaggattg ggaacccctc atcatctgtt ggaatatact attcaaccta caaaagtaac
     3841 gttaaacaac actactttat atttgatatg aatggccaca ccttttatgc cataaaacat
     3901 attgtaagag aataccactc tttttattcc ttctttcctt cttgtacgtt ttttgctgta
     3961 agtaggtcgt ggtgctggtg ttgcagttga aataacttaa aatataaatc ataaaactca
     4021 aacataaact tgactattta tttatttatt aagaaaggaa atataaattg taaattacaa
     4081 caggttccct ttatgcgaag ggatggccaa gcctttgtct caagaagaat ccaccctcat
     4141 tgaaagagca acggctacaa tcaacagcat ccccatctct gaagactaca gcgtcgccag
     4201 cgcagctctc tctagcgacg gccgcatctt cactggtgtc aatgtatatc attttactgg
     4261 gggaccttgc gcagaactcg tggtgctggg cactgctgct gctgcggcag ctggcaacct
     4321 gacttgtatc gtcgcgatcg gaaatgagaa caggggcatc ttgagcccct gcggacggtg
     4381 ccgacaggtt cttctcgatc tgcatcctgg gatcaaagcc atagtgaagg acagtgatgg
     4441 acagccgacg gcagttggga ttcgtgaatt gctgccctct ggttatgtgt gggagggcta
     4501 acccttatac gcaagggagt agatgccgac cgaacaagag ctgatttcga gaacgcctca
     4561 gccagcaact cgcgcgagcc tagcaaggca aatgcgagag aacggcctta cgcttggtgg
     4621 cacagttctc gtccacagtt cgctaagctc gctcggctgg gtcgcgggag ggccggtcgc
     4681 agtgattcag gcccttctgg attgtgttgg tccccagggc acgattgtca tgcccacgca
     4741 ctcgggtgat ctgactgatc ccgcagattg gagatcgccg cccgtgcctg ccgattgggt
     4801 gcagatcagc ctcgaggcca gctagcttga acttgtttat tgcagcttat aatggttaca
     4861 aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt
     4921 gtggtttgtc caaactcatc aatgtatctt atcatgtctg gatcgggctt cctcgctcac
     4981 tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt
     5041 aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca
     5101 gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc
     5161 ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact
     5221 ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct
     5281 gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag
     5341 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca
     5401 cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa
     5461 cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc
     5521 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag
     5581 aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg
     5641 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca
     5701 gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc
     5761 tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag
     5821 gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata
     5881 tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat
     5941 ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg
     6001 ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc
     6061 tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc
     6121 aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc
     6181 gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc
     6241 gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc
     6301 ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa
     6361 gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat
     6421 gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata
     6481 gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca
     6541 tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag
     6601 gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc
     6661 agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc
     6721 aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata
     6781 ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta
     6841 gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgtcta
     6901 agaaaccatt attatcatga cattaaccta taaaaatagg cgtatcacga ggccctttcg
     6961 tctcgcgcgt ttcggtgatg acggtgaaaa cctctgacac atgcagctcc cggagacggt
     7021 cacagcttgt ctgtaagcgg atgccgggag cagacaagcc cgtcagggcg cgtcagcggg
     7081 tgttggcggg tgtcggggct ggcttaacta tgcggcatca gagcagattg tactgagagt
     7141 gcaccatatg cggtgtgaaa taccgcacag atgcgtaagg agaaaatacc gcatcaggcg
     7201 ccattcgcca ttcaggctgc gcaactgttg ggaagggcga tcggtgcggg cctcttcgct
     7261 attacgccag ctggcgaaag ggggatgtgc tgcaaggcga ttaagttggg taacgccagg
     7321 gttttcccag tcacgacgtt gtaaaacgac ggccagtgaa tt
//
