LOCUS       sequence-482290-        8361 bp    DNA     circular SYN 06-AUG-2025
DEFINITION  Expresses mutant NPM1c-TurboID fusion in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8361)
  TITLE     Overexpression of the signaling coordinator GAB2: a missing link in 
            Acute Myeloid Leukemia progression
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 8361)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Aug 6, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     Sequence Label: NPM1c-TurboID-MSCV-IRES-GFP
FEATURES             Location/Qualifiers
     source          1..8361
                     /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"
     regulatory      3441..3450
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             4392..5351
                     /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    5359..5926
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     misc_feature    5376..5928
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(5520..5537)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     5747..5766
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             join(5940..5942,5943..5945,5946..6659)
                     /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(6246..6265)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     6593..6614
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     LTR             6761..7275
                     /label=3' LTR
                     /note="3' long terminal repeat from murine embryonic stem 
                     cell virus"
     protein_bind    7437..7453
                     /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(7461..7467,7468..7485,7486..7491))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7506..7527
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(7644..7661)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(join(7815..8361,1..42))
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7895..7914)
                     /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 taacgaattc gccaccatgg aagattcgat ggacatggac atgagccccc
     3481 tgaggcccca gaactatctt ttcggttgtg aactaaaggc cgacaaagat tatcacttta
     3541 aggtggataa tgatgaaaat gagcaccagt tatctttaag aacggtcagt ttaggggctg
     3601 gtgcaaagga tgagttgcac attgttgaag cagaggcaat gaattacgaa ggcagtccaa
     3661 ttaaagtaac actggcaact ttgaaaatgt ctgtacagcc aacggtttcc cttgggggct
     3721 ttgaaataac accaccagtg gtcttaaggt tgaagtgtgg ttcagggcca gtgcatatta
     3781 gtggacagca cttagtagct gtggaggaag atgcagagtc agaagatgaa gaggaggagg
     3841 atgtgaaact cttaagtata tctggaaagc ggtctgcccc tggaggtggt agcaaggttc
     3901 cacagaaaaa agtaaaactt gctgctgatg aagatgatga cgatgatgat gaagaggatg
     3961 atgatgaaga tgatgatgat gatgattttg atgatgagga agctgaagaa aaagcgccag
     4021 tgaagaaatc tatacgagat actccagcca aaaatgcaca aaagtcaaat cagaatggaa
     4081 aagactcaaa accatcatca acaccaagat caaaaggaca agaatccttc aagaaacagg
     4141 aaaaaactcc taaaacacca aaaggaccta gttctgtaga agacattaaa gcaaaaatgc
     4201 aagcaagtat agaaaaaggt ggttctcttc ccaaagtgga agccaaattc atcaattatg
     4261 tgaagaattg cttccggatg actgaccaag aggctattca agatctctgt ctggcagtgg
     4321 aggaagtctc tttaagaaaa ggatccggtg gaggtggttc tggcggaggc ggctctggag
     4381 gaggaggaag tatgaaagac aatactgtgc ctctgaagct gatcgctctc ctggctaatg
     4441 gcgagttcca tagtggcgaa cagctgggag aaaccctggg catgtccagg gccgctatca
     4501 acaagcacat tcagactctg cgcgactggg gcgtggacgt gttcaccgtg cccggaaagg
     4561 gctactctct gcccgagcct atcccgctgc tgaacgctaa acagattctg ggacagctgg
     4621 acggcgggag cgtggcagtc ctgcctgtgg tcgactccac caatcagtac ctgctggatc
     4681 gaatcggcga gctgaagagt ggggatgctt gcattgcaga atatcagcag gcagggagag
     4741 gaagcagagg gaggaaatgg ttctctcctt ttggagctaa cctgtacctg agtatgtttt
     4801 ggcgcctgaa gcggggacca gcagcaatcg gcctgggccc ggtcatcgga attgtcatgg
     4861 cagaagcgct gcgaaagctg ggagcagaca aggtgcgagt caaatggccc aatgacctgt
     4921 atctgcagga tagaaagctg gcaggcatcc tggtggagct ggccggaata acaggcgatg
     4981 ctgcacagat cgtcattggc gccgggatta acgtggctat gaggcgcgtg gaggaaagcg
     5041 tggtcaatca gggctggatc acactgcagg aagcagggat taacctggac aggaatactc
     5101 tggccgctac gctgatccga gagctgcggg cagccctgga actgttcgag caggaaggcc
     5161 tggctccata tctgccacgg tgggagaagc tggataactt catcaataga cccgtgaagc
     5221 tgatcattgg ggacaaagag attttcggga ttagccgggg gattgataaa cagggagccc
     5281 tgctgctgga acaggacgga gttatcaaac cctggatggg cggagaaatc agtctgcggt
     5341 ctgccgaaaa gtagcaattc cgcccccccc ccctaacgtt actggccgaa gccgcttgga
     5401 ataaggccgg tgtgcgtttg tctatatgtt attttccacc atattgccgt cttttggcaa
     5461 tgtgagggcc cggaaacctg gccctgtctt cttgacgagc attcctaggg gtctttcccc
     5521 tctcgccaaa ggaatgcaag gtctgttgaa tgtcgtgaag gaagcagttc ctctggaagc
     5581 ttcttgaaga caaacaacgt ctgtagcgac cctttgcagg cagcggaacc ccccacctgg
     5641 cgacaggtgc ctctgcggcc aaaagccacg tgtataagat acacctgcaa aggcggcaca
     5701 accccagtgc cacgttgtga gttggatagt tgtggaaaga gtcaaatggc tctcctcaag
     5761 cgtattcaac aaggggctga aggatgccca gaaggtaccc cattgtatgg gatctgatct
     5821 ggggcctcgg tgcacatgct ttacatgtgt ttagtcgagg ttaaaaaaac gtctaggccc
     5881 cccgaaccac ggggacgtgg ttttcctttg aaaaacacga tgataatatg gccacaacca
     5941 tggtgagcaa gggcgaggag ctgttcaccg gggtggtgcc catcctggtc gagctggacg
     6001 gcgacgtaaa cggccacaag ttcagcgtgt ctggcgaggg cgagggcgat gccacctacg
     6061 gcaagctgac cctgaagttc atctgcacca ccggcaagct gcccgtgccc tggcccaccc
     6121 tcgtgaccac cctgacctac ggcgtgcagt gcttcagccg ctaccccgac cacatgaagc
     6181 agcacgactt cttcaagtcc gccatgcccg aaggctacgt ccaggagcgc accatcttct
     6241 tcaaggacga cggcaactac aagacccgcg ccgaggtgaa gttcgagggc gacaccctgg
     6301 tgaaccgcat cgagctgaag ggcatcgact tcaaggagga cggcaacatc ctggggcaca
     6361 agctggagta caactacaac agccacaacg tctatatcat ggccgacaag cagaagaacg
     6421 gcatcaaggc gaacttcaag atccgccaca acatcgagga cggcagcgtg cagctcgccg
     6481 accactacca gcagaacacc cccatcggcg acggccccgt gctgctgccc gacaaccact
     6541 acctgagcac ccagtccgcc ctgagcaaag accccaacga gaagcgcgat cacatggtcc
     6601 tgctggagtt cgtgaccgcc gccgggatca ctctcggcat ggacgagctg tacaagtaat
     6661 gaattaatta agaattatca agcttatcga taccgtcgac ctgcagccaa gcttatcgat
     6721 aaaataaaag attttattta gtctccagaa aaagggggga atgaaagacc ccacctgtag
     6781 gtttggcaag ctagcttaag taacgccatt ttgcaaggca tggaaaatac ataactgaga
     6841 atagagaagt tcagatcaag gttaggaaca gagagacagc agaatatggg ccaaacagga
     6901 tatctgtggt aagcagttcc tgccccggct cagggccaag aacagatggt ccccagatgc
     6961 ggtcccgccc tcagcagttt ctagagaacc atcagatgtt tccagggtgc cccaaggacc
     7021 tgaaatgacc ctgtgcctta tttgaactaa ccaatcagtt cgcttctcgc ttctgttcgc
     7081 gcgcttctgc tccccgagct caataaaaga gcccacaacc cctcactcgg cgcgccagtc
     7141 ctccgataga ctgcgtcgcc cgggtacccg tgtatccaat aaaccctctt gcagttgcat
     7201 ccgacttgtg gtctcgctgt tccttgggag ggtctcctct gagtgattga ctacccgtca
     7261 gcgggggtct ttcatgggta acagtttctt gaagttggag aacaacattc tgagggtagg
     7321 agtcgaatat taagtaatcc tgactcaatt agccactgtt ttgaatccac atactccaat
     7381 actcctgaaa tagttcatta tggacagcgc agaaagagct ggggagaatt gtgaaattgt
     7441 tatccgctca caattccaca caacatacga gccggaagca taaagtgtaa agcctggggt
     7501 gcctaatgag tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg
     7561 ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg
     7621 cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg
     7681 cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat
     7741 aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc
     7801 gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc
     7861 tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga
     7921 agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt
     7981 ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg
     8041 taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc
     8101 gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg
     8161 gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc
     8221 ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg
     8281 ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc
     8341 gctggtagcg gtggtttttt t
//
