LOCUS       sequence-327845-        7693 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express the Hyperactive catalytic domain of Drosophila ADAR and 
            linker at the 5' end of this domain, in the MSCV-IRES-GFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7693)
  AUTHORS   Nguyen DTT, Lu Y, Chu KL, Yang X, Park SM, Choo ZN, Chin CR, Prieto 
            C, Schurer A, Barin E, Savino AM, Gourkanti S, Patel P, Vu LP, 
            Leslie CS, Kharas MG
  TITLE     HyperTRIBE uncovers increased MUSASHI-2 RNA binding activity and 
            differential regulation in leukemic stem cells.
  JOURNAL   Nat Commun. 2020 Apr 24;11(1):2026. doi: 10.1038/s41467-020-15814-8.
  PUBMED    32332729
REFERENCE   2  (bases 1 to 7693)
  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: "Nat 
            Commun."; date: "2020-04-24"; volume: "11(1):2026. doi"; pages: " 
            10.1038/s41467-020-15814-8"
COMMENT     Sequence Label: HyperADAR control
FEATURES             Location/Qualifiers
     source          1..7693
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(join(528..1319,1320..1388))
                     /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     1151..1170
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1389..1493)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     1561..1579
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(1617..1639)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     1739..1758
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     1952..1974
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     2102..2121
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     LTR             2321..2837
                     /label=5' LTR
                     /note="5' long terminal repeat from murine embryonic stem 
                     cell virus"
     misc_feature    2901..3242
                     /label=MESV Psi
                     /note="packaging signal of murine embryonic stem cell 
                     virus"
     CDS             3309..3725
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     3655..3677
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     CDS             4953..5000
                     /codon_start=1
                     /product="two tandem FLAG(R) epitope tags"
                     /label=2xFLAG
                     /translation="DYKDDDDKDYKDDDDK"
     misc_feature    5008..5573
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     misc_feature    5023..5575
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(5167..5184)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     5394..5413
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             join(5587..5589,5590..5592,5593..6306)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     ANFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(5632..5653)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(5893..5912)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     6240..6261
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     LTR             6408..6922
                     /label=3' LTR
                     /note="3' long terminal repeat from murine embryonic stem 
                     cell virus"
     protein_bind    7084..7100
                     /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(7108..7114,7115..7132,7133..7138))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7153..7174
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(7291..7308)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(join(7462..7693,1..357))
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7542..7561)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta
       61 tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca
      121 gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag
      181 tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag
      241 ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt
      301 agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa
      361 gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg
      421 attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga
      481 agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta
      541 atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc
      601 cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg
      661 ataccgcgag acccacgctc accggctcca gatttatcag caataaacca gccagccgga
      721 agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt
      781 tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt
      841 gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc
      901 caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc
      961 ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca
     1021 gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag
     1081 tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg
     1141 tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa
     1201 cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa
     1261 cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga
     1321 gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga
     1381 atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg
     1441 agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt
     1501 ccccgaaaag tgccacctga cgtctaagaa accattatta tcatgacatt aacctataaa
     1561 aataggcgta tcacgaggcc ctttcgtctc gcgcgtttcg gtgatgacgg tgaaaacctc
     1621 tgacacatgc agctcccgga gacggtcaca gcttgtctgt aagcggatgc cgggagcaga
     1681 caagcccgtc agggcgcgtc agcgggtgtt ggcgggtgtc ggggctggct taactatgcg
     1741 gcatcagagc agattgtact gagagtgcac catatgcggt gtgaaatacc gcacagatgc
     1801 gtaaggagaa aataccgcat caggcgccat tcgccattca ggctgcgcaa ctgttgggaa
     1861 gggcgatcgg tgcgggcctc ttcgctatta cgccagctgg cgaaaggggg atgtgctgca
     1921 aggcgattaa gttgggtaac gccagggttt tcccagtcac gacgttgtaa aacgacggcg
     1981 caaggaatgg tgcatgcaag gagatggcgc ccaacagtcc cccggccacg gggcctgcca
     2041 ccatacccac gccgaaacaa gcgctcatga gcccgaagtg gcgagcccga tcttccccat
     2101 cggtgatgtc ggcgatatag gcgccagcaa ccgcacctgt ggcgccggtg atgccggcca
     2161 cgatgcgtcc ggcgtagagg cgattagtcc aatttgttaa agacaggata tcagtggtcc
     2221 aggctctagt tttgactcaa caatatcacc agctgaagcc tatagagtac gagccataga
     2281 taaaataaaa gattttattt agtctccaga aaaagggggg aatgaaagac cccacctgta
     2341 ggtttggcaa gctagcttaa gtaacgccat tttgcaaggc atggaaaata cataactgag
     2401 aatagagaag ttcagatcaa ggttaggaac agagagacag cagaatatgg gccaaacagg
     2461 atatctgtgg taagcagttc ctgccccggc tcagggccaa gaacagatgg tccccagatg
     2521 cggtcccgcc ctcagcagtt tctagagaac catcagatgt ttccagggtg ccccaaggac
     2581 ctgaaatgac cctgtgcctt atttgaacta accaatcagt tcgcttctcg cttctgttcg
     2641 cgcgcttctg ctccccgagc tcaataaaag agcccacaac ccctcactcg gcgcgccagt
     2701 cctccgatag actgcgtcgc ccgggtaccc gtattcccaa taaagcctct tgctgtttgc
     2761 atccgaatcg tggactcgct gatccttggg agggtctcct cagattgatt gactgcccac
     2821 ctcgggggtc tttcatttgg aggttccacc gagatttgga gacccctgcc cagggaccac
     2881 cgaccccccc gccgggaggt aagctggcca gcggtcgttt cgtgtctgtc tctgtctttg
     2941 tgcgtgtttg tgccggcatc taatgtttgc gcctgcgtct gtactagtta gctaactagc
     3001 tctgtatctg gcggacccgt ggtggaactg acgagttctg aacacccggc cgcaaccctg
     3061 ggagacgtcc cagggacttt gggggccgtt tttgtggccc gacctgagga agggagtcga
     3121 tgtggaatcc gaccccgtca ggatatgtgg ttctggtagg agacgagaac ctaaaacagt
     3181 tcccgcctcc gtctgaattt ttgctttcgg tttggaaccg aagccgcgcg tcttgtctgc
     3241 tgcagcgctg cagcatcgtt ctgtgttgtc tctgtctgac tgtgtttctg tatttgtctg
     3301 aaaattaggg ccagactgtt accactccct taagtttgac cttaggtcac tggaaagatg
     3361 tcgagcggat cgctcacaac cagtcggtag atgtcaagaa gagacgttgg gttaccttct
     3421 gctctgcaga atggccaacc tttaacgtcg gatggccgcg agacggcacc tttaaccgag
     3481 acctcatcac ccaggttaag atcaaggtct tttcacctgg cccgcatgga cacccagacc
     3541 aggtccccta catcgtgacc tgggaagcct tggcttttga cccccctccc tgggtcaagc
     3601 cctttgtaca ccctaagcct ccgcctcctc ttcctccatc cgccccgtct ctcccccttg
     3661 aacctcctcg ttcgaccccg cctcgatcct ccctttatcc agccctcact ccttctctag
     3721 gcgccggaat tagatctctc gagatgtatt cccccatggt ggtgccccag aagaacgtcc
     3781 ccctccccat tgacgacaaa agcagcagca tggagctccc ccagatccac gctgacacaa
     3841 ttggcaggct ggtgctggaa aaattcatgg aggtgatcaa aggacaagag gcctacagca
     3901 ggaggaaagt gctggctggc attgtgatga ccgagaatat gaacttctgc gaggccaagg
     3961 tgatcagcgt gtccaccggc accaaatgcg tctccggcga acacatgagc gtcaacggcg
     4021 ccgtgctgaa tgattcccac gccgagattg tctccagaag gtgcctcctc aagtacctct
     4081 acgcccagct ggacctgcag tgcaaccaag ctaccgccta ccagagcatc ttcgtcagaa
     4141 acaccgatgg ccagtacccc tataagctga agagcggagt gcacttccac ctctatatca
     4201 ataccgctcc ttgcggcgac gctaggatct tcagccctca cgagaacgac acaggcgtgg
     4261 acaagcatcc caacaggaaa gccaggggcc agctgaggac caagatcgag agcggccagg
     4321 gaaccatccc cgtgaagagc agcgatggca tccagacctg ggatggagtc ctccagggcc
     4381 agagactgct gaccatgtcc tgtagcgaca agatcgccag gtggaatatc gtgggcattc
     4441 aaggctccct cctgtccagc atcattgaac ccgtgtacct gcactccatc gtgctgggct
     4501 ccctgctgca ccccgaacac atgtataggg ctgtgtgcgg cagaattgag aagagcatcc
     4561 agggcctccc tcccccctac cacctcaaca aacccaggct ggccctcgtc accagcgccg
     4621 agcccagaaa ccaggccaaa gcccccaact tcggcatcaa ctggaccatc ggagataccg
     4681 agctggaggt ggtgaacagc ctcaccggaa ggaccattgg cggccaggtg tccaggatca
     4741 caaagcaggc cttcttcgtg aaatacggct tcctcatggc caacctgcct ggcatcctgg
     4801 tgagaaaagt caccacagat tacggccaga caaaagccaa cgtgaaggac taccagattg
     4861 ccaagctgga actgttcagc gctttcaaga gagaagacct gggcagctgg ctgaagaaac
     4921 ccatcgaaca ggatgagttc ggactggctg aggactacaa agacgatgac gacaaggatt
     4981 ataaagacga cgacgacaag tgagaattcc gccccccccc taacgttact ggccgaagcc
     5041 gcttggaata aggccggtgt gcgtttgtct atatgttatt ttccaccata ttgccgtctt
     5101 ttggcaatgt gagggcccgg aaacctggcc ctgtcttctt gacgagcatt cctaggggtc
     5161 tttcccctct cgccaaagga atgcaaggtc tgttgaatgt cgtgaaggaa gcagttcctc
     5221 tggaagcttc ttgaagacaa acaacgtctg tagcgaccct ttgcaggcag cggaaccccc
     5281 cacctggcga caggtgcctc tgcggccaaa agccacgtgt ataagataca cctgcaaagg
     5341 cggcacaacc ccagtgccac gttgtgagtt ggatagttgt ggaaagagtc aaatggctct
     5401 cctcaagcgt attcaacaag gggctgaagg atgcccagaa ggtaccccat tgtatgggat
     5461 ctgatctggg gcctcggtgc acatgcttta catgtgttta gtcgaggtta aaaaaacgtc
     5521 taggcccccc gaaccacggg gacgtggttt tcctttgaaa aacacgatga taatatggcc
     5581 acaaccatgg tgagcaaggg cgaggagctg ttcaccgggg tggtgcccat cctggtcgag
     5641 ctggacggcg acgtaaacgg ccacaagttc agcgtgtctg gcgagggcga gggcgatgcc
     5701 acctacggca agctgaccct gaagttcatc tgcaccaccg gcaagctgcc cgtgccctgg
     5761 cccaccctcg tgaccaccct gacctacggc gtgcagtgct tcagccgcta ccccgaccac
     5821 atgaagcagc acgacttctt caagtccgcc atgcccgaag gctacgtcca ggagcgcacc
     5881 atcttcttca aggacgacgg caactacaag acccgcgccg aggtgaagtt cgagggcgac
     5941 accctggtga accgcatcga gctgaagggc atcgacttca aggaggacgg caacatcctg
     6001 gggcacaagc tggagtacaa ctacaacagc cacaacgtct atatcatggc cgacaagcag
     6061 aagaacggca tcaaggcgaa cttcaagatc cgccacaaca tcgaggacgg cagcgtgcag
     6121 ctcgccgacc actaccagca gaacaccccc atcggcgacg gccccgtgct gctgcccgac
     6181 aaccactacc tgagcaccca gtccgccctg agcaaagacc ccaacgagaa gcgcgatcac
     6241 atggtcctgc tggagttcgt gaccgccgcc gggatcactc tcggcatgga cgagctgtac
     6301 aagtaatgaa ttaattaaga attatcaagc ttatcgatac cgtcgacctg cagccaagct
     6361 tatcgataaa ataaaagatt ttatttagtc tccagaaaaa ggggggaatg aaagacccca
     6421 cctgtaggtt tggcaagcta gcttaagtaa cgccattttg caaggcatgg aaaatacata
     6481 actgagaata gagaagttca gatcaaggtt aggaacagag agacagcaga atatgggcca
     6541 aacaggatat ctgtggtaag cagttcctgc cccggctcag ggccaagaac agatggtccc
     6601 cagatgcggt cccgccctca gcagtttcta gagaaccatc agatgtttcc agggtgcccc
     6661 aaggacctga aatgaccctg tgccttattt gaactaacca atcagttcgc ttctcgcttc
     6721 tgttcgcgcg cttctgctcc ccgagctcaa taaaagagcc cacaacccct cactcggcgc
     6781 gccagtcctc cgatagactg cgtcgcccgg gtacccgtgt atccaataaa ccctcttgca
     6841 gttgcatccg acttgtggtc tcgctgttcc ttgggagggt ctcctctgag tgattgacta
     6901 cccgtcagcg ggggtctttc atgggtaaca gtttcttgaa gttggagaac aacattctga
     6961 gggtaggagt cgaatattaa gtaatcctga ctcaattagc cactgttttg aatccacata
     7021 ctccaatact cctgaaatag ttcattatgg acagcgcaga aagagctggg gagaattgtg
     7081 aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc
     7141 ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt
     7201 ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg
     7261 cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt
     7321 tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc
     7381 aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa
     7441 aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa
     7501 tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc
     7561 ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc
     7621 cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag
     7681 ttcggtgtag gtc
//
