LOCUS       sequence-327843-        8674 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express the fusion protein of human MSI2 and the catalytic dead 
            domain of Drosophila ADAR.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8674)
  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 8674)
  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-DCD
FEATURES             Location/Qualifiers
     source          1..8674
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    252..268
                     /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(276..282,283..300,301..306))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    321..342
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(459..476)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(630..1218)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(710..729)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1389..2180,2181..2249))
                     /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     2012..2031
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2250..2354)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     2422..2440
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(2478..2500)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     2600..2619
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2813..2835
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     2963..2982
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     LTR             3182..3698
                     /label=5' LTR
                     /note="5' long terminal repeat from murine embryonic stem 
                     cell virus"
     misc_feature    3762..4103
                     /label=MESV Psi
                     /note="packaging signal of murine embryonic stem cell 
                     virus"
     CDS             4170..4586
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     4516..4538
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     CDS             6795..6842
                     /codon_start=1
                     /product="two tandem FLAG(R) epitope tags"
                     /label=2xFLAG
                     /translation="DYKDDDDKDYKDDDDK"
     misc_feature    6850..7415
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     misc_feature    6865..7417
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(7009..7026)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     7236..7255
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             join(7429..7431,7432..7434,7435..8148)
                     /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(7474..7495)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(7735..7754)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     8082..8103
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     LTR             join(8250..8674,1..90)
                     /label=3' LTR
                     /note="3' long terminal repeat from murine embryonic stem 
                     cell virus"
ORIGIN
        1 ctcttgcagt tgcatccgac ttgtggtctc gctgttcctt gggagggtct cctctgagtg
       61 attgactacc cgtcagcggg ggtctttcat gggtaacagt ttcttgaagt tggagaacaa
      121 cattctgagg gtaggagtcg aatattaagt aatcctgact caattagcca ctgttttgaa
      181 tccacatact ccaatactcc tgaaatagtt cattatggac agcgcagaaa gagctgggga
      241 gaattgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg aagcataaag
      301 tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt gcgctcactg
      361 cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg
      421 gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc
      481 tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc
      541 acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg
      601 aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat
      661 cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag
      721 gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga
      781 tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg
      841 tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt
      901 cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac
      961 gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc
     1021 ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt
     1081 ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc
     1141 ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc
     1201 agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg
     1261 aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag
     1321 atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg
     1381 tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt
     1441 tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca
     1501 tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca
     1561 gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc
     1621 tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt
     1681 ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg
     1741 gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc
     1801 aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg
     1861 ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga
     1921 tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga
     1981 ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta
     2041 aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg
     2101 ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact
     2161 ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata
     2221 agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt
     2281 tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa
     2341 ataggggttc cgcgcacatt tccccgaaaa gtgccacctg acgtctaaga aaccattatt
     2401 atcatgacat taacctataa aaataggcgt atcacgaggc cctttcgtct cgcgcgtttc
     2461 ggtgatgacg gtgaaaacct ctgacacatg cagctcccgg agacggtcac agcttgtctg
     2521 taagcggatg ccgggagcag acaagcccgt cagggcgcgt cagcgggtgt tggcgggtgt
     2581 cggggctggc ttaactatgc ggcatcagag cagattgtac tgagagtgca ccatatgcgg
     2641 tgtgaaatac cgcacagatg cgtaaggaga aaataccgca tcaggcgcca ttcgccattc
     2701 aggctgcgca actgttggga agggcgatcg gtgcgggcct cttcgctatt acgccagctg
     2761 gcgaaagggg gatgtgctgc aaggcgatta agttgggtaa cgccagggtt ttcccagtca
     2821 cgacgttgta aaacgacggc gcaaggaatg gtgcatgcaa ggagatggcg cccaacagtc
     2881 ccccggccac ggggcctgcc accataccca cgccgaaaca agcgctcatg agcccgaagt
     2941 ggcgagcccg atcttcccca tcggtgatgt cggcgatata ggcgccagca accgcacctg
     3001 tggcgccggt gatgccggcc acgatgcgtc cggcgtagag gcgattagtc caatttgtta
     3061 aagacaggat atcagtggtc caggctctag ttttgactca acaatatcac cagctgaagc
     3121 ctatagagta cgagccatag ataaaataaa agattttatt tagtctccag aaaaaggggg
     3181 gaatgaaaga ccccacctgt aggtttggca agctagctta agtaacgcca ttttgcaagg
     3241 catggaaaat acataactga gaatagagaa gttcagatca aggttaggaa cagagagaca
     3301 gcagaatatg ggccaaacag gatatctgtg gtaagcagtt cctgccccgg ctcagggcca
     3361 agaacagatg gtccccagat gcggtcccgc cctcagcagt ttctagagaa ccatcagatg
     3421 tttccagggt gccccaagga cctgaaatga ccctgtgcct tatttgaact aaccaatcag
     3481 ttcgcttctc gcttctgttc gcgcgcttct gctccccgag ctcaataaaa gagcccacaa
     3541 cccctcactc ggcgcgccag tcctccgata gactgcgtcg cccgggtacc cgtattccca
     3601 ataaagcctc ttgctgtttg catccgaatc gtggactcgc tgatccttgg gagggtctcc
     3661 tcagattgat tgactgccca cctcgggggt ctttcatttg gaggttccac cgagatttgg
     3721 agacccctgc ccagggacca ccgacccccc cgccgggagg taagctggcc agcggtcgtt
     3781 tcgtgtctgt ctctgtcttt gtgcgtgttt gtgccggcat ctaatgtttg cgcctgcgtc
     3841 tgtactagtt agctaactag ctctgtatct ggcggacccg tggtggaact gacgagttct
     3901 gaacacccgg ccgcaaccct gggagacgtc ccagggactt tgggggccgt ttttgtggcc
     3961 cgacctgagg aagggagtcg atgtggaatc cgaccccgtc aggatatgtg gttctggtag
     4021 gagacgagaa cctaaaacag ttcccgcctc cgtctgaatt tttgctttcg gtttggaacc
     4081 gaagccgcgc gtcttgtctg ctgcagcgct gcagcatcgt tctgtgttgt ctctgtctga
     4141 ctgtgtttct gtatttgtct gaaaattagg gccagactgt taccactccc ttaagtttga
     4201 ccttaggtca ctggaaagat gtcgagcgga tcgctcacaa ccagtcggta gatgtcaaga
     4261 agagacgttg ggttaccttc tgctctgcag aatggccaac ctttaacgtc ggatggccgc
     4321 gagacggcac ctttaaccga gacctcatca cccaggttaa gatcaaggtc ttttcacctg
     4381 gcccgcatgg acacccagac caggtcccct acatcgtgac ctgggaagcc ttggcttttg
     4441 acccccctcc ctgggtcaag ccctttgtac accctaagcc tccgcctcct cttcctccat
     4501 ccgccccgtc tctccccctt gaacctcctc gttcgacccc gcctcgatcc tccctttatc
     4561 cagccctcac tccttctcta ggcgccggaa ttagatctct cgagatggaa gccaacggca
     4621 gccagggcac ctccggctcc gccaatgaca gccaacacga ccccggaaag atgttcatcg
     4681 gcggcctcag ctggcagacc agccccgata gcctgaggga ctacttctcc aaattcggcg
     4741 aaattaggga gtgcatggtg atgagagacc ccacaaccaa gaggtccagg ggcttcggct
     4801 ttgtcacctt cgccgatccc gcttccgtgg acaaggtcct gggccagccc catcatgagc
     4861 tggactccaa gacaatcgac cccaaagtgg cctttcccag gagagcccag cccaagatgg
     4921 tgaccaggac caagaagatc ttcgtgggcg gcctgagcgc taacaccgtg gtggaggacg
     4981 tcaagcaata cttcgagcag ttcggaaagg tcgaagacgc catgctgatg ttcgacaaga
     5041 caaccaatag gcacagaggc ttcggcttcg tcaccttcga gaacgaggat gtcgtcgaga
     5101 aggtgtgcga gattcacttc cacgaaatta acaataagat ggtcgagtgc aagaaggctc
     5161 agcctaaaga ggtcatgttc ccccccggca caagaggcag agctaggggc ctcccctata
     5221 caatggacgc cttcatgctg ggcatgggca tgctgggcta ccccaacttt gtggctacat
     5281 acggcagggg ctatcccggc ttcgccccct cctatggcta ccagttcccc ggattccccg
     5341 ctgctgctta cggacccgtg gccgccgccg ccgtggctgc cgccagaggc agcggatcca
     5401 atcccgctag gcctggcggc tttcccggcg ctaatagccc tggacccgtc gctgacctgt
     5461 atggacccgc ctcccaggac agcggcgtgg gaaactatat ttccgccgct agcccccaac
     5521 ccggatccgg attcggccat ggcattgccg gccccctgat cgctaccgcc ttcaccaatg
     5581 gctaccacta ttcccccatg gtggtgcccc agaagaacgt ccccctcccc attgacgaca
     5641 aaagcagcag catggagctc ccccagatcc acgctgacac aattggcagg ctggtgctgg
     5701 aaaaattcat ggaggtgatc aaaggacaag aggcctacag caggaggaaa gtgctggctg
     5761 gcattgtgat gaccgagaat atgaacttct gcgaggccaa ggtgatcagc gtgtccaccg
     5821 gcaccaaatg cgtctccggc gaacacatga gcgtcaacgg cgccgtgctg aatgattccc
     5881 acgccgccat tgtctccaga aggtgcctcc tcaagtacct ctacgcccag ctggacctgc
     5941 agtgcaacca agctaccgcc taccagagca tcttcgtcag aaacaccgat ggccagtacc
     6001 cctataagct gaagagcgga gtgcacttcc acctctatat caataccgct ccttgcggcg
     6061 acgctaggat cttcagccct cacgagagcg acacaggcgt ggacaagcat cccaacagga
     6121 aagccagggg ccagctgagg accaagatcg agagcggcca gggaaccatc cccgtgaaga
     6181 gcagcgatgg catccagacc tgggatggag tcctccaggg ccagagactg ctgaccatgt
     6241 cctgtagcga caagatcgcc aggtggaata tcgtgggcat tcaaggctcc ctcctgtcca
     6301 gcatcattga acccgtgtac ctgcactcca tcgtgctggg ctccctgctg caccccgaac
     6361 acatgtatag ggctgtgtgc ggcagaattg agaagagcat ccagggcctc cctcccccct
     6421 accacctcaa caaacccagg ctggccctcg tcaccagcgc cgagcccaga aaccaggcca
     6481 aagcccccaa cttcggcatc aactggacca tcggagatac cgagctggag gtggtgaaca
     6541 gcctcaccgg aaggaccatt ggcggccagg tgtccaggat cacaaagcag gccttcttcg
     6601 tgaaatacgg cttcctcatg gccaacctgc ctggcatcct ggtgagaaaa gtcaccacag
     6661 attacggcca gacaaaagcc aacgtgaagg actaccagat tgccaagctg gaactgttca
     6721 gcgctttcaa gagagaagac ctgggcagct ggctgaagaa acccatcgaa caggatgagt
     6781 tcggactggc tgaggactac aaagacgatg acgacaagga ttataaagac gacgacgaca
     6841 agtgagaatt ccgccccccc cctaacgtta ctggccgaag ccgcttggaa taaggccggt
     6901 gtgcgtttgt ctatatgtta ttttccacca tattgccgtc ttttggcaat gtgagggccc
     6961 ggaaacctgg ccctgtcttc ttgacgagca ttcctagggg tctttcccct ctcgccaaag
     7021 gaatgcaagg tctgttgaat gtcgtgaagg aagcagttcc tctggaagct tcttgaagac
     7081 aaacaacgtc tgtagcgacc ctttgcaggc agcggaaccc cccacctggc gacaggtgcc
     7141 tctgcggcca aaagccacgt gtataagata cacctgcaaa ggcggcacaa ccccagtgcc
     7201 acgttgtgag ttggatagtt gtggaaagag tcaaatggct ctcctcaagc gtattcaaca
     7261 aggggctgaa ggatgcccag aaggtacccc attgtatggg atctgatctg gggcctcggt
     7321 gcacatgctt tacatgtgtt tagtcgaggt taaaaaaacg tctaggcccc ccgaaccacg
     7381 gggacgtggt tttcctttga aaaacacgat gataatatgg ccacaaccat ggtgagcaag
     7441 ggcgaggagc tgttcaccgg ggtggtgccc atcctggtcg agctggacgg cgacgtaaac
     7501 ggccacaagt tcagcgtgtc tggcgagggc gagggcgatg ccacctacgg caagctgacc
     7561 ctgaagttca tctgcaccac cggcaagctg cccgtgccct ggcccaccct cgtgaccacc
     7621 ctgacctacg gcgtgcagtg cttcagccgc taccccgacc acatgaagca gcacgacttc
     7681 ttcaagtccg ccatgcccga aggctacgtc caggagcgca ccatcttctt caaggacgac
     7741 ggcaactaca agacccgcgc cgaggtgaag ttcgagggcg acaccctggt gaaccgcatc
     7801 gagctgaagg gcatcgactt caaggaggac ggcaacatcc tggggcacaa gctggagtac
     7861 aactacaaca gccacaacgt ctatatcatg gccgacaagc agaagaacgg catcaaggcg
     7921 aacttcaaga tccgccacaa catcgaggac ggcagcgtgc agctcgccga ccactaccag
     7981 cagaacaccc ccatcggcga cggccccgtg ctgctgcccg acaaccacta cctgagcacc
     8041 cagtccgccc tgagcaaaga ccccaacgag aagcgcgatc acatggtcct gctggagttc
     8101 gtgaccgccg ccgggatcac tctcggcatg gacgagctgt acaagtaatg aattaattaa
     8161 gaattatcaa gcttatcgat accgtcgacc tgcagccaag cttatcgata aaataaaaga
     8221 ttttatttag tctccagaaa aaggggggaa tgaaagaccc cacctgtagg tttggcaagc
     8281 tagcttaagt aacgccattt tgcaaggcat ggaaaataca taactgagaa tagagaagtt
     8341 cagatcaagg ttaggaacag agagacagca gaatatgggc caaacaggat atctgtggta
     8401 agcagttcct gccccggctc agggccaaga acagatggtc cccagatgcg gtcccgccct
     8461 cagcagtttc tagagaacca tcagatgttt ccagggtgcc ccaaggacct gaaatgaccc
     8521 tgtgccttat ttgaactaac caatcagttc gcttctcgct tctgttcgcg cgcttctgct
     8581 ccccgagctc aataaaagag cccacaaccc ctcactcggc gcgccagtcc tccgatagac
     8641 tgcgtcgccc gggtacccgt gtatccaata aacc
//
