LOCUS       sequence-309423-        7322 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  expresses mADAR2 shortest transcript, puromycin selection.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7322)
  AUTHORS   Abukar A, Wipplinger M, Hariharan A, Sun S, Ronner M, Sculco M, 
            Okonska A, Kresoja-Rakic J, Rehrauer H, Qi W, Beusechem VWV, 
            Felley-Bosco E
  TITLE     Double-Stranded RNA Structural Elements Holding the Key to 
            Translational Regulation in Cancer: The Case of Editing in 
            RNA-Binding Motif Protein 8A.
  JOURNAL   Cells. 2021 Dec 15;10(12). pii: cells10123543. doi: 
            10.3390/cells10123543.
  PUBMED    34944051
REFERENCE   2  (bases 1 to 7322)
  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: "Cells."; 
            date: "2021-12-15"; volume: "10(12). pii: cells10123543. doi"; 
            pages: " 10.3390/cells10123543"
COMMENT     Sequence Label: pCI-Puro_mouse_Adar2_Tv1
FEATURES             Location/Qualifiers
     source          1..7322
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        138..517
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        518..729
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     680..700
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     726..750
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     intron          890..1022
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     1005..1024
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     promoter        1067..1085
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     polyA_signal    3277..3398
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3288..3307)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3342..3361
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     rep_origin      3584..4039
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3671..3690)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3881..3902
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     enhancer        4156..4347
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4179..4199)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4197..4494
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     CDS             4506..5105
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(4506..4525)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     5002..5022
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    5171..5219
                     /note="synthetic polyadenylation signal"
     primer_bind     complement(5260..5279)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5379..5401
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5439..5457)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5525..5629
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5630..5698,5699..6490)
                     /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(5848..5867)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6661..7249
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7150..7169
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tcaatattgg ccattagcca tattattcat tggttatata gcataaatca atattggcta
       61 ttggccattg catacgttgt atctatatca taatatgtac atttatattg gctcatgtcc
      121 aatatgaccg ccatgttggc attgattatt gactagttat taatagtaat caattacggg
      181 gtcattagtt catagcccat atatggagtt ccgcgttaca taacttacgg taaatggccc
      241 gcctggctga ccgcccaacg acccccgccc attgacgtca ataatgacgt atgttcccat
      301 agtaacgcca atagggactt tccattgacg tcaatgggtg gagtatttac ggtaaactgc
      361 ccacttggca gtacatcaag tgtatcatat gccaagtccg ccccctattg acgtcaatga
      421 cggtaaatgg cccgcctggc attatgccca gtacatgacc ttacgggact ttcctacttg
      481 gcagtacatc tacgtattag tcatcgctat taccatggtg atgcggtttt ggcagtacac
      541 caatgggcgt ggatagcggt ttgactcacg gggatttcca agtctccacc ccattgacgt
      601 caatgggagt ttgttttggc accaaaatca acgggacttt ccaaaatgtc gtaacaactg
      661 cgatcgcccg ccccgttgac gcaaatgggc ggtaggcgtg tacggtggga ggtctatata
      721 agcagagctc gtttagtgaa ccgtcagatc actagaagct ttattgcggt agtttatcac
      781 agttaaattg ctaacgcagt cagtgcttct gacacaacag tctcgaactt aagctgcagt
      841 gactctctta aggtagcctt gcagaagttg gtcgtgaggc actgggcagg taagtatcaa
      901 ggttacaaga caggtttaag gagaccaata gaaactgggc ttgtcgagac agagaagact
      961 cttgcgtttc tgataggcac ctattggtct tactgacatc cactttgcct ttctctccac
     1021 aggtgtccac tcccagttca attacagctc ttaaggctag agtacttaat acgactcact
     1081 ataggctagc atggatatag aagatgaaga gaatatgagt tccagcagca ctgatattaa
     1141 agaaaaccgc aatctggaca acatgccccc caaggacagc agcacacctg ggcctggcga
     1201 gggtattccg ctctccaacg ggggtggtgg tagcaccagc aggaagcggc ccctggagga
     1261 gggcagcaat ggtcactcca agtaccgcct gaagaagcga aggaaaacac cagggcctgt
     1321 tctgcccaag aacgccctga tgcagctgaa cgagatcaaa cctggcttac agtacatgct
     1381 gctgtcccag acaggacccg tgcatgcacc tctgtttgtc atgtctgtgg aggtaaacgg
     1441 gcaggttttt gagggctctg gccctacaaa gaaaaaggca aaactccatg ctgctgagaa
     1501 ggccctgagg tcttttgtcc agtttcccaa cgcctccgag gcccacctag ccatgggaag
     1561 gaccctctct gtgaacacag acttcacgtc tgaccaggct gacttccctg acacactctt
     1621 caatggcttc gagactccag acaagtcaga gccacccttc tacgtaggct ccaacgggga
     1681 tgactcgttc agctcaagtg gagatgtcag cctatcggcc tccccagtgc ctgccagcct
     1741 tacccagcct cctctgccca tcccaccacc attcccaccc ccaagtggga agaatcccgt
     1801 gatgatcttg aatgagctac gcccagggct gaagtatgac ttcctctctg agagtgggga
     1861 gagccacgcc aagagcttcg tcatgtccgt ggtggtagat ggccagttct ttgagggctc
     1921 aggaagaaac aagaagcttg ccaaggcccg ggctgcacag tctgccttgg ctactgtctt
     1981 caatttgcac ttggaccaaa caccatctcg ccagcctgtc ctcagtgagg gtcttcagtt
     2041 gcatttgcca caggtattgg cagatgctgt ctcccgcctg gtcctgggta agttcagtga
     2101 cctgacagac aacttttcct cccctcatgc acgaaggaaa gtgctctctg gagtagtgat
     2161 gaccacaggt acagatgtca aagatgccaa ggtgataagt gtttcgacag ggacgaagtg
     2221 tatcaacggt gaatacatga gtgaccgtgg cctcgcactc aatgactgcc acgcagagat
     2281 aatctcccga aggtccctgc tcaggtttct ttatgcacag ctcgagcttt atttaaataa
     2341 caaagaagac cagaaaaagt ccatatttca gaagtcagag cggggtgggt tccggctgaa
     2401 ggataccgtg cagttccacc tgtacatcag cacctcgccc tgcggagacg ccagaatatt
     2461 ctctccccac gagcccgtgc tagaggaacc agcagataga catccgaatc gcaaagcaag
     2521 gggacagcta cggacgaaaa tagagtctgg cgaggggaca atccctgtgc gctcaaatgc
     2581 cagcatccag acctgggacg gggtgctgca gggggaacgg ctgctcacca tgtcctgcag
     2641 tgacaagata gcacgctgga acgtggtggg catccagggg tccctgctca gcattttcgt
     2701 ggaacccatc tacttctcca gcatcatctt gggcagcctg taccacgggg accacctctc
     2761 cagggccatg taccagcgga tctccaacat agaggacctg ccaccgctct acactctcaa
     2821 caagcccctg ctcagcggca tcagcaatgc agaggcacgg cagccaggga aggcacccaa
     2881 cttcagtgtc aactggacag tgggtgacgc caccattgag gtcatcaatg ccacaacagg
     2941 gaaggatgag cttggccgcc catcccgcct gtgtaagcac gcgctgtact gtcgctggat
     3001 gcgtgtacac ggcaaggttc ccccccacct gctgcgcacc aagatcacta agcctaccac
     3061 ataccacgag tccaagctgg cagcgaggga gtaccaggct gccaaggccc gtctgttcac
     3121 tgccttcatc aaggcggggc tgggcgcctg ggtggagaag cccacagagc aggaccagtt
     3181 ctccttcact ccctgagaat tcacgcgtgg tacctctaga gtcgacccgg gcggccgctt
     3241 ccctttagtg agggttaatg cttcgagcag acatgataag atacattgat gagtttggac
     3301 aaaccacaac tagaatgcag tgaaaaaaat gctttatttg tgaaatttgt gatgctattg
     3361 ctttatttgt aaccattata agctgcaata aacaagttaa caacaacaat tgcattcatt
     3421 ttatgtttca ggttcagggg gagatgtggg aggtttttta aagcaagtaa aacctctaca
     3481 aatgtggtaa aatccgataa ggatcgatcc gggctggcgt aatagcgaag aggcccgcac
     3541 cgatcgccct tcccaacagt tgcgcagcct gaatggcgaa tggacgcgcc ctgtagcggc
     3601 gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact tgccagcgcc
     3661 ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc cggctttccc
     3721 cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagtgcttt acggcacctc
     3781 gaccccaaaa aacttgatta gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg
     3841 gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt gttccaaact
     3901 ggaacaacac tcaaccctat ctcggtctat tcttttgatt tataagggat tttgccgatt
     3961 tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa ttttaacaaa
     4021 atattaacgc ttacaatttc ctgatgcggt attttctcct tacgcatctg tgcggtattt
     4081 cacaccgcat acgcggatct gcgcagcacc atggcctgaa ataacctctg aaagaggaac
     4141 ttggttaggt accttctgag gcggaaagaa ccagctgtgg aatgtgtgtc agttagggtg
     4201 tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc
     4261 agcaaccagg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca
     4321 tctcaattag tcagcaacca tagtcccgcc cctaactccg cccatcccgc ccctaactcc
     4381 gcccagttcc gcccattctc cgccccatgg ctgactaatt ttttttattt atgcagaggc
     4441 cgaggccgcc tcggcctctg agctattcca gaagtagtga ggaggctttt ttggaggagc
     4501 ttaccatgac cgagtacaag cccacggtgc gcctcgccac ccgcgacgac gtccccaggg
     4561 ccgtacgcac cctcgccgcc gcgttcgccg actaccccgc cacgcgccac accgtcgatc
     4621 cggaccgcca catcgagcgg gtcaccgagc tgcaagaact cttcctcacg cgcgtcgggc
     4681 tcgacatcgg caaggtgtgg gtcgcggacg acggcgccgc ggtggcggtc tggaccacgc
     4741 cggagagcgt cgaagcgggg gcggtgttcg ccgagatcgg cccgcgcatg gccgagttga
     4801 gcggttcccg gctggccgcg cagcaacaga tggaaggcct cctggcgccg caccggccca
     4861 aggagcccgc gtggttcctg gccaccgtcg gcgtctcgcc cgaccaccag ggcaagggtc
     4921 tgggcagcgc cgtcgtgctc cccggagtgg aggcggccga gcgcgccggg gtgcccgcct
     4981 tcctggagac ctccgcgccc cgcaacctcc ccttctacga gcggctcggc ttcaccgtca
     5041 ccgccgacgt cgaggtgccc gaaggaccgc gcacctggtg catgacccgc aagcccggtg
     5101 cctgacgccc gccccacgac ccgcagcgcc cgaccgaaag gagcgcacga ccccatgcat
     5161 cgatggccgc aataaaatat ctttattttc attacatctg tgtgttggtt ttttgtgtga
     5221 atcgatagcg ataaggatcc gcgtatggtg cactctcagt acaatctgct ctgatgccgc
     5281 atagttaagc cagccccgac acccgccaac acccgctgac gcgccctgac gggcttgtct
     5341 gctcccggca tccgcttaca gacaagctgt gaccgtctcc gggagctgca tgtgtcagag
     5401 gttttcaccg tcatcaccga aacgcgcgag acgaaagggc ctcgtgatac gcctattttt
     5461 ataggttaat gtcatgataa taatggtttc ttagacgtca ggtggcactt ttcggggaaa
     5521 tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat
     5581 gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagta tgagtattca
     5641 acatttccgt gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca
     5701 cccagaaacg ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac gagtgggtta
     5761 catcgaactg gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgttt
     5821 tccaatgatg agcactttta aagttctgct atgtggcgcg gtattatccc gtattgacgc
     5881 cgggcaagag caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtactc
     5941 accagtcaca gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc
     6001 cataaccatg agtgataaca ctgcggccaa cttacttctg acaacgatcg gaggaccgaa
     6061 ggagctaacc gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga
     6121 accggagctg aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgtagcaat
     6181 ggcaacaacg ttgcgcaaac tattaactgg cgaactactt actctagctt cccggcaaca
     6241 attaatagac tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc
     6301 ggctggctgg tttattgctg ataaatctgg agccggtgag cgtgggtctc gcggtatcat
     6361 tgcagcactg gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag
     6421 tcaggcaact atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa
     6481 gcattggtaa ctgtcagacc aagtttactc atatatactt tagattgatt taaaacttca
     6541 tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc
     6601 ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc
     6661 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
     6721 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
     6781 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
     6841 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
     6901 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
     6961 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
     7021 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
     7081 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
     7141 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
     7201 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
     7261 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgg ctcgacagat
     7321 ct
//
