LOCUS       sequence-479874-        8360 bp    DNA     circular SYN 07-AUG-2025
DEFINITION  Expresses TurboID-NPM1c fusion in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8360)
  AUTHORS   Kramer MH, Richardson SN, Li Y, Yin T, Helton NM, George DR, Cai M, 
            Ramakrishnan SM, Katerndahl CDS, Miller CA, Ley TJ
  TITLE     Overexpression of the signaling-coordinator GAB2 can play an 
            important role in Acute Myeloid Leukemia progression
  JOURNAL   J Clin Invest. 2025.
REFERENCE   2  (bases 1 to 8360)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Aug 7, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "J Clin 
            Invest. 2025."
COMMENT     Sequence Label: TurboID-NPM1c-MSCV-IRES-GFP
FEATURES             Location/Qualifiers
     source          1..8360
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(join(213..1004,1005..1073))
                     /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"
     promoter        complement(1074..1178)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     1246..1264
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(1302..1324)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     1424..1443
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     1637..1659
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     1787..1806
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     LTR             2006..2522
                     /label=5' LTR
                     /note="5' long terminal repeat from murine embryonic stem 
                     cell virus"
     misc_feature    2586..2927
                     /label=MESV Psi
                     /note="packaging signal of murine embryonic stem cell 
                     virus"
     CDS             2994..3410
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     3340..3362
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     3378..3394
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     CDS             3447..4406
                     /codon_start=1
                     /product="mutant of the E. coli biotin ligase BioID 
                     engineered for improved catalytic efficiency (Branon et 
                     al., 2018)"
                     /label=TurboID
                     /translation="MKDNTVPLKLIALLANGEFHSGEQLGETLGMSRAAINKHIQTLRD
                     WGVDVFTVPGKGYSLPEPIPLLNAKQILGQLDGGSVAVLPVVDSTNQYLLDRIGELKSG
                     DACIAEYQQAGRGSRGRKWFSPFGANLYLSMFWRLKRGPAAIGLGPVIGIVMAEALRKL
                     GADKVRVKWPNDLYLQDRKLAGILVELAGITGDAAQIVIGAGINVAMRRVEESVVNQGW
                     ITLQEAGINLDRNTLAATLIRELRAALELFEQEGLAPYLPRWEKLDNFINRPVKLIIGD
                     KEIFGISRGIDKQGALLLEQDGVIKPWMGGEISLRSAEK"
     misc_feature    5375..5927
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(5519..5536)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     5746..5765
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             join(5939..5941,5942..5944,5945..6658)
                     /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(6245..6264)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     6592..6613
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     LTR             6760..7274
                     /label=3' LTR
                     /note="3' long terminal repeat from murine embryonic stem 
                     cell virus"
     protein_bind    7436..7452
                     /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(7460..7466,7467..7484,7485..7490))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7505..7526
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(7643..7660)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(join(7814..8360,1..42))
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7894..7913)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt
       61 tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga
      121 ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc
      181 taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct
      241 atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata
      301 actacgatac gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca
      361 cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga
      421 agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga
      481 gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg
      541 gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga
      601 gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt
      661 gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct
      721 cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca
      781 ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat
      841 accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga
      901 aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc
      961 aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg
     1021 caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc
     1081 ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt
     1141 gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca
     1201 cctgacgtct aagaaaccat tattatcatg acattaacct ataaaaatag gcgtatcacg
     1261 aggccctttc gtctcgcgcg tttcggtgat gacggtgaaa acctctgaca catgcagctc
     1321 ccggagacgg tcacagcttg tctgtaagcg gatgccggga gcagacaagc ccgtcagggc
     1381 gcgtcagcgg gtgttggcgg gtgtcggggc tggcttaact atgcggcatc agagcagatt
     1441 gtactgagag tgcaccatat gcggtgtgaa ataccgcaca gatgcgtaag gagaaaatac
     1501 cgcatcaggc gccattcgcc attcaggctg cgcaactgtt gggaagggcg atcggtgcgg
     1561 gcctcttcgc tattacgcca gctggcgaaa gggggatgtg ctgcaaggcg attaagttgg
     1621 gtaacgccag ggttttccca gtcacgacgt tgtaaaacga cggcgcaagg aatggtgcat
     1681 gcaaggagat ggcgcccaac agtcccccgg ccacggggcc tgccaccata cccacgccga
     1741 aacaagcgct catgagcccg aagtggcgag cccgatcttc cccatcggtg atgtcggcga
     1801 tataggcgcc agcaaccgca cctgtggcgc cggtgatgcc ggccacgatg cgtccggcgt
     1861 agaggcgatt agtccaattt gttaaagaca ggatatcagt ggtccaggct ctagttttga
     1921 ctcaacaata tcaccagctg aagcctatag agtacgagcc atagataaaa taaaagattt
     1981 tatttagtct ccagaaaaag gggggaatga aagaccccac ctgtaggttt ggcaagctag
     2041 cttaagtaac gccattttgc aaggcatgga aaatacataa ctgagaatag agaagttcag
     2101 atcaaggtta ggaacagaga gacagcagaa tatgggccaa acaggatatc tgtggtaagc
     2161 agttcctgcc ccggctcagg gccaagaaca gatggtcccc agatgcggtc ccgccctcag
     2221 cagtttctag agaaccatca gatgtttcca gggtgcccca aggacctgaa atgaccctgt
     2281 gccttatttg aactaaccaa tcagttcgct tctcgcttct gttcgcgcgc ttctgctccc
     2341 cgagctcaat aaaagagccc acaacccctc actcggcgcg ccagtcctcc gatagactgc
     2401 gtcgcccggg tacccgtatt cccaataaag cctcttgctg tttgcatccg aatcgtggac
     2461 tcgctgatcc ttgggagggt ctcctcagat tgattgactg cccacctcgg gggtctttca
     2521 tttggaggtt ccaccgagat ttggagaccc ctgcccaggg accaccgacc cccccgccgg
     2581 gaggtaagct ggccagcggt cgtttcgtgt ctgtctctgt ctttgtgcgt gtttgtgccg
     2641 gcatctaatg tttgcgcctg cgtctgtact agttagctaa ctagctctgt atctggcgga
     2701 cccgtggtgg aactgacgag ttctgaacac ccggccgcaa ccctgggaga cgtcccaggg
     2761 actttggggg ccgtttttgt ggcccgacct gaggaaggga gtcgatgtgg aatccgaccc
     2821 cgtcaggata tgtggttctg gtaggagacg agaacctaaa acagttcccg cctccgtctg
     2881 aatttttgct ttcggtttgg aaccgaagcc gcgcgtcttg tctgctgcag cgctgcagca
     2941 tcgttctgtg ttgtctctgt ctgactgtgt ttctgtattt gtctgaaaat tagggccaga
     3001 ctgttaccac tcccttaagt ttgaccttag gtcactggaa agatgtcgag cggatcgctc
     3061 acaaccagtc ggtagatgtc aagaagagac gttgggttac cttctgctct gcagaatggc
     3121 caacctttaa cgtcggatgg ccgcgagacg gcacctttaa ccgagacctc atcacccagg
     3181 ttaagatcaa ggtcttttca cctggcccgc atggacaccc agaccaggtc ccctacatcg
     3241 tgacctggga agccttggct tttgaccccc ctccctgggt caagcccttt gtacacccta
     3301 agcctccgcc tcctcttcct ccatccgccc cgtctctccc ccttgaacct cctcgttcga
     3361 ccccgcctcg atcctccctt tatccagccc tcactccttc tctaggcgcc ggaattagat
     3421 ctctcgaggt taaccaattc gccaccatga aagacaatac tgtgcctctg aagctgatcg
     3481 ctctcctggc taatggcgag ttccatagtg gcgaacagct gggagaaacc ctgggcatgt
     3541 ccagggccgc tatcaacaag cacattcaga ctctgcgcga ctggggcgtg gacgtgttca
     3601 ccgtgcccgg aaagggctac tctctgcccg agcctatccc gctgctgaac gctaaacaga
     3661 ttctgggaca gctggacggc gggagcgtgg cagtcctgcc tgtggtcgac tccaccaatc
     3721 agtacctgct ggatcgaatc ggcgagctga agagtgggga tgcttgcatt gcagaatatc
     3781 agcaggcagg gagaggaagc agagggagga aatggttctc tccttttgga gctaacctgt
     3841 acctgagtat gttttggcgc ctgaagcggg gaccagcagc aatcggcctg ggcccggtca
     3901 tcggaattgt catggcagaa gcgctgcgaa agctgggagc agacaaggtg cgagtcaaat
     3961 ggcccaatga cctgtatctg caggatagaa agctggcagg catcctggtg gagctggccg
     4021 gaataacagg cgatgctgca cagatcgtca ttggcgccgg gattaacgtg gctatgaggc
     4081 gcgtggagga aagcgtggtc aatcagggct ggatcacact gcaggaagca gggattaacc
     4141 tggacaggaa tactctggcc gctacgctga tccgagagct gcgggcagcc ctggaactgt
     4201 tcgagcagga aggcctggct ccatatctgc cacggtggga gaagctggat aacttcatca
     4261 atagacccgt gaagctgatc attggggaca aagagatttt cgggattagc cgggggattg
     4321 ataaacaggg agccctgctg ctggaacagg acggagttat caaaccctgg atgggcggag
     4381 aaatcagtct gcggtctgcc gaaaagggtg gaggtggttc tggcggaggc ggctctggag
     4441 gaggaggaag tggatccatg gaagattcga tggacatgga catgagcccc ctgaggcccc
     4501 agaactatct tttcggttgt gaactaaagg ccgacaaaga ttatcacttt aaggtggata
     4561 atgatgaaaa tgagcaccag ttatctttaa gaacggtcag tttaggggct ggtgcaaagg
     4621 atgagttgca cattgttgaa gcagaggcaa tgaattacga aggcagtcca attaaagtaa
     4681 cactggcaac tttgaaaatg tctgtacagc caacggtttc ccttgggggc tttgaaataa
     4741 caccaccagt ggtcttaagg ttgaagtgtg gttcagggcc agtgcatatt agtggacagc
     4801 acttagtagc tgtggaggaa gatgcagagt cagaagatga agaggaggag gatgtgaaac
     4861 tcttaagtat atctggaaag cggtctgccc ctggaggtgg tagcaaggtt ccacagaaaa
     4921 aagtaaaact tgctgctgat gaagatgatg acgatgatga tgaagaggat gatgatgaag
     4981 atgatgatga tgatgatttt gatgatgagg aagctgaaga aaaagcgcca gtgaagaaat
     5041 ctatacgaga tactccagcc aaaaatgcac aaaagtcaaa tcagaatgga aaagactcaa
     5101 aaccatcatc aacaccaaga tcaaaaggac aagaatcctt caagaaacag gaaaaaactc
     5161 ctaaaacacc aaaaggacct agttctgtag aagacattaa agcaaaaatg caagcaagta
     5221 tagaaaaagg tggttctctt cccaaagtgg aagccaaatt catcaattat gtgaagaatt
     5281 gcttccggat gactgaccaa gaggctattc aagatctctg tctggcagtg gaggaagtct
     5341 ctttaagaaa ataggaattc gccccccccc cctaacgtta ctggccgaag ccgcttggaa
     5401 taaggccggt gtgcgtttgt ctatatgtta ttttccacca tattgccgtc ttttggcaat
     5461 gtgagggccc ggaaacctgg ccctgtcttc ttgacgagca ttcctagggg tctttcccct
     5521 ctcgccaaag gaatgcaagg tctgttgaat gtcgtgaagg aagcagttcc tctggaagct
     5581 tcttgaagac aaacaacgtc tgtagcgacc ctttgcaggc agcggaaccc cccacctggc
     5641 gacaggtgcc tctgcggcca aaagccacgt gtataagata cacctgcaaa ggcggcacaa
     5701 ccccagtgcc acgttgtgag ttggatagtt gtggaaagag tcaaatggct ctcctcaagc
     5761 gtattcaaca aggggctgaa ggatgcccag aaggtacccc attgtatggg atctgatctg
     5821 gggcctcggt gcacatgctt tacatgtgtt tagtcgaggt taaaaaaacg tctaggcccc
     5881 ccgaaccacg gggacgtggt tttcctttga aaaacacgat gataatatgg ccacaaccat
     5941 ggtgagcaag ggcgaggagc tgttcaccgg ggtggtgccc atcctggtcg agctggacgg
     6001 cgacgtaaac ggccacaagt tcagcgtgtc tggcgagggc gagggcgatg ccacctacgg
     6061 caagctgacc ctgaagttca tctgcaccac cggcaagctg cccgtgccct ggcccaccct
     6121 cgtgaccacc ctgacctacg gcgtgcagtg cttcagccgc taccccgacc acatgaagca
     6181 gcacgacttc ttcaagtccg ccatgcccga aggctacgtc caggagcgca ccatcttctt
     6241 caaggacgac ggcaactaca agacccgcgc cgaggtgaag ttcgagggcg acaccctggt
     6301 gaaccgcatc gagctgaagg gcatcgactt caaggaggac ggcaacatcc tggggcacaa
     6361 gctggagtac aactacaaca gccacaacgt ctatatcatg gccgacaagc agaagaacgg
     6421 catcaaggcg aacttcaaga tccgccacaa catcgaggac ggcagcgtgc agctcgccga
     6481 ccactaccag cagaacaccc ccatcggcga cggccccgtg ctgctgcccg acaaccacta
     6541 cctgagcacc cagtccgccc tgagcaaaga ccccaacgag aagcgcgatc acatggtcct
     6601 gctggagttc gtgaccgccg ccgggatcac tctcggcatg gacgagctgt acaagtaatg
     6661 aattaattaa gaattatcaa gcttatcgat accgtcgacc tgcagccaag cttatcgata
     6721 aaataaaaga ttttatttag tctccagaaa aaggggggaa tgaaagaccc cacctgtagg
     6781 tttggcaagc tagcttaagt aacgccattt tgcaaggcat ggaaaataca taactgagaa
     6841 tagagaagtt cagatcaagg ttaggaacag agagacagca gaatatgggc caaacaggat
     6901 atctgtggta agcagttcct gccccggctc agggccaaga acagatggtc cccagatgcg
     6961 gtcccgccct cagcagtttc tagagaacca tcagatgttt ccagggtgcc ccaaggacct
     7021 gaaatgaccc tgtgccttat ttgaactaac caatcagttc gcttctcgct tctgttcgcg
     7081 cgcttctgct ccccgagctc aataaaagag cccacaaccc ctcactcggc gcgccagtcc
     7141 tccgatagac tgcgtcgccc gggtacccgt gtatccaata aaccctcttg cagttgcatc
     7201 cgacttgtgg tctcgctgtt ccttgggagg gtctcctctg agtgattgac tacccgtcag
     7261 cgggggtctt tcatgggtaa cagtttcttg aagttggaga acaacattct gagggtagga
     7321 gtcgaatatt aagtaatcct gactcaatta gccactgttt tgaatccaca tactccaata
     7381 ctcctgaaat agttcattat ggacagcgca gaaagagctg gggagaattg tgaaattgtt
     7441 atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg
     7501 cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg
     7561 gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc
     7621 gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc
     7681 ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata
     7741 acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg
     7801 cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct
     7861 caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa
     7921 gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc
     7981 tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt
     8041 aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg
     8101 ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg
     8161 cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct
     8221 tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc
     8281 tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg
     8341 ctggtagcgg tggttttttt
//
