LOCUS       sequence-327834-        8734 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express the fusion protein of human MSI2 and the hyperactive 
            catalytic domain of Drosophila ADAR.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8734)
  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 8734)
  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: MSI2-ADA
FEATURES             Location/Qualifiers
     source          1..8734
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    580..921
                     /label=MESV Psi
                     /note="packaging signal of murine embryonic stem cell 
                     virus"
     CDS             988..1404
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     1334..1356
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     CDS             3613..3660
                     /codon_start=1
                     /product="two tandem FLAG(R) epitope tags"
                     /label=2xFLAG
                     /translation="DYKDDDDKDYKDDDDK"
     misc_feature    3668..4233
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     misc_feature    3683..4235
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(3827..3844)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     4054..4073
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             join(4247..4249,4250..4252,4253..4966)
                     /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(4292..4313)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(4553..4572)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4900..4921
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     LTR             5128..5642
                     /label=3' LTR
                     /note="3' long terminal repeat from murine embryonic stem 
                     cell virus"
     protein_bind    5804..5820
                     /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(5828..5834,5835..5852,5853..5858))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5873..5894
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6011..6028)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6182..6770)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6262..6281)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6941..7732,7733..7801))
                     /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     7564..7583
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7802..7906)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     7974..7992
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(8030..8052)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     8152..8171
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     8365..8387
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     8515..8534
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     LTR             join(8734,1..516)
                     /label=5' LTR
                     /note="5' long terminal repeat from murine embryonic stem 
                     cell virus"
ORIGIN
        1 atgaaagacc ccacctgtag gtttggcaag ctagcttaag taacgccatt ttgcaaggca
       61 tggaaaatac ataactgaga atagagaagt tcagatcaag gttaggaaca gagagacagc
      121 agaatatggg ccaaacagga tatctgtggt aagcagttcc tgccccggct cagggccaag
      181 aacagatggt ccccagatgc ggtcccgccc tcagcagttt ctagagaacc atcagatgtt
      241 tccagggtgc cccaaggacc tgaaatgacc ctgtgcctta tttgaactaa ccaatcagtt
      301 cgcttctcgc ttctgttcgc gcgcttctgc tccccgagct caataaaaga gcccacaacc
      361 cctcactcgg cgcgccagtc ctccgataga ctgcgtcgcc cgggtacccg tattcccaat
      421 aaagcctctt gctgtttgca tccgaatcgt ggactcgctg atccttggga gggtctcctc
      481 agattgattg actgcccacc tcgggggtct ttcatttgga ggttccaccg agatttggag
      541 acccctgccc agggaccacc gacccccccg ccgggaggta agctggccag cggtcgtttc
      601 gtgtctgtct ctgtctttgt gcgtgtttgt gccggcatct aatgtttgcg cctgcgtctg
      661 tactagttag ctaactagct ctgtatctgg cggacccgtg gtggaactga cgagttctga
      721 acacccggcc gcaaccctgg gagacgtccc agggactttg ggggccgttt ttgtggcccg
      781 acctgaggaa gggagtcgat gtggaatccg accccgtcag gatatgtggt tctggtagga
      841 gacgagaacc taaaacagtt cccgcctccg tctgaatttt tgctttcggt ttggaaccga
      901 agccgcgcgt cttgtctgct gcagcgctgc agcatcgttc tgtgttgtct ctgtctgact
      961 gtgtttctgt atttgtctga aaattagggc cagactgtta ccactccctt aagtttgacc
     1021 ttaggtcact ggaaagatgt cgagcggatc gctcacaacc agtcggtaga tgtcaagaag
     1081 agacgttggg ttaccttctg ctctgcagaa tggccaacct ttaacgtcgg atggccgcga
     1141 gacggcacct ttaaccgaga cctcatcacc caggttaaga tcaaggtctt ttcacctggc
     1201 ccgcatggac acccagacca ggtcccctac atcgtgacct gggaagcctt ggcttttgac
     1261 ccccctccct gggtcaagcc ctttgtacac cctaagcctc cgcctcctct tcctccatcc
     1321 gccccgtctc tcccccttga acctcctcgt tcgaccccgc ctcgatcctc cctttatcca
     1381 gccctcactc cttctctagg cgccggaatt agatctctcg agatggaagc caacggcagc
     1441 cagggcacct ccggctccgc caatgacagc caacacgacc ccggaaagat gttcatcggc
     1501 ggcctcagct ggcagaccag ccccgatagc ctgagggact acttctccaa attcggcgaa
     1561 attagggagt gcatggtgat gagagacccc acaaccaaga ggtccagggg cttcggcttt
     1621 gtcaccttcg ccgatcccgc ttccgtggac aaggtcctgg gccagcccca tcatgagctg
     1681 gactccaaga caatcgaccc caaagtggcc tttcccagga gagcccagcc caagatggtg
     1741 accaggacca agaagatctt cgtgggcggc ctgagcgcta acaccgtggt ggaggacgtc
     1801 aagcaatact tcgagcagtt cggaaaggtc gaagacgcca tgctgatgtt cgacaagaca
     1861 accaataggc acagaggctt cggcttcgtc accttcgaga acgaggatgt cgtcgagaag
     1921 gtgtgcgaga ttcacttcca cgaaattaac aataagatgg tcgagtgcaa gaaggctcag
     1981 cctaaagagg tcatgttccc ccccggcaca agaggcagag ctaggggcct cccctataca
     2041 atggacgcct tcatgctggg catgggcatg ctgggctacc ccaactttgt ggctacatac
     2101 ggcaggggct atcccggctt cgccccctcc tatggctacc agttccccgg attccccgct
     2161 gctgcttacg gacccgtggc cgccgccgcc gtggctgccg ccagaggcag cggatccaat
     2221 cccgctaggc ctggcggctt tcccggcgct aatagccctg gacccgtcgc tgacctgtat
     2281 ggacccgcct cccaggacag cggcgtggga aactatattt ccgccgctag cccccaaccc
     2341 ggatccggat tcggccatgg cattgccggc cccctgatcg ctaccgcctt caccaatggc
     2401 taccactatt cccccatggt ggtgccccag aagaacgtcc ccctccccat tgacgacaaa
     2461 agcagcagca tggagctccc ccagatccac gctgacacaa ttggcaggct ggtgctggaa
     2521 aaattcatgg aggtgatcaa aggacaagag gcctacagca ggaggaaagt gctggctggc
     2581 attgtgatga ccgagaatat gaacttctgc gaggccaagg tgatcagcgt gtccaccggc
     2641 accaaatgcg tctccggcga acacatgagc gtcaacggcg ccgtgctgaa tgattcccac
     2701 gccgagattg tctccagaag gtgcctcctc aagtacctct acgcccagct ggacctgcag
     2761 tgcaaccaag ctaccgccta ccagagcatc ttcgtcagaa acaccgatgg ccagtacccc
     2821 tataagctga agagcggagt gcacttccac ctctatatca ataccgctcc ttgcggcgac
     2881 gctaggatct tcagccctca cgagaacgac acaggcgtgg acaagcatcc caacaggaaa
     2941 gccaggggcc agctgaggac caagatcgag agcggccagg gaaccatccc cgtgaagagc
     3001 agcgatggca tccagacctg ggatggagtc ctccagggcc agagactgct gaccatgtcc
     3061 tgtagcgaca agatcgccag gtggaatatc gtgggcattc aaggctccct cctgtccagc
     3121 atcattgaac ccgtgtacct gcactccatc gtgctgggct ccctgctgca ccccgaacac
     3181 atgtataggg ctgtgtgcgg cagaattgag aagagcatcc agggcctccc tcccccctac
     3241 cacctcaaca aacccaggct ggccctcgtc accagcgccg agcccagaaa ccaggccaaa
     3301 gcccccaact tcggcatcaa ctggaccatc ggagataccg agctggaggt ggtgaacagc
     3361 ctcaccggaa ggaccattgg cggccaggtg tccaggatca caaagcaggc cttcttcgtg
     3421 aaatacggct tcctcatggc caacctgcct ggcatcctgg tgagaaaagt caccacagat
     3481 tacggccaga caaaagccaa cgtgaaggac taccagattg ccaagctgga actgttcagc
     3541 gctttcaaga gagaagacct gggcagctgg ctgaagaaac ccatcgaaca ggatgagttc
     3601 ggactggctg aggactacaa agacgatgac gacaaggatt ataaagacga cgacgacaag
     3661 tgagaattcc gccccccccc taacgttact ggccgaagcc gcttggaata aggccggtgt
     3721 gcgtttgtct atatgttatt ttccaccata ttgccgtctt ttggcaatgt gagggcccgg
     3781 aaacctggcc ctgtcttctt gacgagcatt cctaggggtc tttcccctct cgccaaagga
     3841 atgcaaggtc tgttgaatgt cgtgaaggaa gcagttcctc tggaagcttc ttgaagacaa
     3901 acaacgtctg tagcgaccct ttgcaggcag cggaaccccc cacctggcga caggtgcctc
     3961 tgcggccaaa agccacgtgt ataagataca cctgcaaagg cggcacaacc ccagtgccac
     4021 gttgtgagtt ggatagttgt ggaaagagtc aaatggctct cctcaagcgt attcaacaag
     4081 gggctgaagg atgcccagaa ggtaccccat tgtatgggat ctgatctggg gcctcggtgc
     4141 acatgcttta catgtgttta gtcgaggtta aaaaaacgtc taggcccccc gaaccacggg
     4201 gacgtggttt tcctttgaaa aacacgatga taatatggcc acaaccatgg tgagcaaggg
     4261 cgaggagctg ttcaccgggg tggtgcccat cctggtcgag ctggacggcg acgtaaacgg
     4321 ccacaagttc agcgtgtctg gcgagggcga gggcgatgcc acctacggca agctgaccct
     4381 gaagttcatc tgcaccaccg gcaagctgcc cgtgccctgg cccaccctcg tgaccaccct
     4441 gacctacggc gtgcagtgct tcagccgcta ccccgaccac atgaagcagc acgacttctt
     4501 caagtccgcc atgcccgaag gctacgtcca ggagcgcacc atcttcttca aggacgacgg
     4561 caactacaag acccgcgccg aggtgaagtt cgagggcgac accctggtga accgcatcga
     4621 gctgaagggc atcgacttca aggaggacgg caacatcctg gggcacaagc tggagtacaa
     4681 ctacaacagc cacaacgtct atatcatggc cgacaagcag aagaacggca tcaaggcgaa
     4741 cttcaagatc cgccacaaca tcgaggacgg cagcgtgcag ctcgccgacc actaccagca
     4801 gaacaccccc atcggcgacg gccccgtgct gctgcccgac aaccactacc tgagcaccca
     4861 gtccgccctg agcaaagacc ccaacgagaa gcgcgatcac atggtcctgc tggagttcgt
     4921 gaccgccgcc gggatcactc tcggcatgga cgagctgtac aagtaatgaa ttaattaaga
     4981 attatcaagc ttatcgatac cgtcsayskr crrscwrkmy wakyratgaa ttaattaaga
     5041 attatcaagc ttatcgatac cgtcgacctg cagccaagct tatcgataaa ataaaagatt
     5101 ttatttagtc tccagaaaaa ggggggaatg aaagacccca cctgtaggtt tggcaagcta
     5161 gcttaagtaa cgccattttg caaggcatgg aaaatacata actgagaata gagaagttca
     5221 gatcaaggtt aggaacagag agacagcaga atatgggcca aacaggatat ctgtggtaag
     5281 cagttcctgc cccggctcag ggccaagaac agatggtccc cagatgcggt cccgccctca
     5341 gcagtttcta gagaaccatc agatgtttcc agggtgcccc aaggacctga aatgaccctg
     5401 tgccttattt gaactaacca atcagttcgc ttctcgcttc tgttcgcgcg cttctgctcc
     5461 ccgagctcaa taaaagagcc cacaacccct cactcggcgc gccagtcctc cgatagactg
     5521 cgtcgcccgg gtacccgtgt atccaataaa ccctcttgca gttgcatccg acttgtggtc
     5581 tcgctgttcc ttgggagggt ctcctctgag tgattgacta cccgtcagcg ggggtctttc
     5641 atgggtaaca gtttcttgaa gttggagaac aacattctga gggtaggagt cgaatattaa
     5701 gtaatcctga ctcaattagc cactgttttg aatccacata ctccaatact cctgaaatag
     5761 ttcattatgg acagcgcaga aagagctggg gagaattgtg aaattgttat ccgctcacaa
     5821 ttccacacaa catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga
     5881 gctaactcac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt
     5941 gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct
     6001 cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat
     6061 cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga
     6121 acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt
     6181 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt
     6241 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc
     6301 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa
     6361 gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct
     6421 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta
     6481 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg
     6541 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc
     6601 ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta
     6661 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg
     6721 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt
     6781 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg
     6841 tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta
     6901 aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg
     6961 aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg
     7021 tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc
     7081 gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg
     7141 agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg
     7201 aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag
     7261 gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat
     7321 caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc
     7381 cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc
     7441 ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa
     7501 ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac
     7561 gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt
     7621 cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc
     7681 gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa
     7741 caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca
     7801 tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat
     7861 acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa
     7921 aagtgccacc tgacgtctaa gaaaccatta ttatcatgac attaacctat aaaaataggc
     7981 gtatcacgag gccctttcgt ctcgcgcgtt tcggtgatga cggtgaaaac ctctgacaca
     8041 tgcagctccc ggagacggtc acagcttgtc tgtaagcgga tgccgggagc agacaagccc
     8101 gtcagggcgc gtcagcgggt gttggcgggt gtcggggctg gcttaactat gcggcatcag
     8161 agcagattgt actgagagtg caccatatgc ggtgtgaaat accgcacaga tgcgtaagga
     8221 gaaaataccg catcaggcgc cattcgccat tcaggctgcg caactgttgg gaagggcgat
     8281 cggtgcgggc ctcttcgcta ttacgccagc tggcgaaagg gggatgtgct gcaaggcgat
     8341 taagttgggt aacgccaggg ttttcccagt cacgacgttg taaaacgacg gcgcaaggaa
     8401 tggtgcatgc aaggagatgg cgcccaacag tcccccggcc acggggcctg ccaccatacc
     8461 cacgccgaaa caagcgctca tgagcccgaa gtggcgagcc cgatcttccc catcggtgat
     8521 gtcggcgata taggcgccag caaccgcacc tgtggcgccg gtgatgccgg ccacgatgcg
     8581 tccggcgtag aggcgattag tccaatttgt taaagacagg atatcagtgg tccaggctct
     8641 agttttgact caacaatatc accagctgaa gcctatagag tacgagccat agataaaata
     8701 aaagatttta tttagtctcc agaaaaaggg ggga
//
