LOCUS       sequence-414095-        7479 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Retroviral vector for cloning a gene of interest with a 
            glycine-serine linker and TurboID on the C-terminus.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7479)
  AUTHORS   Day RB, Hickman JA, Xu Z, Katerndahl CD, Ferraro F, Ramakrishnan SM,
            Erdmann-Gilmore P, Sprung RW, Mi Y, Townsend RR, Miller CA, Ley TJ
  TITLE     Proteogenomic analysis reveals cytoplasmic sequestration of RUNX1 by
            the acute myeloid leukemia-initiating CBFB::MYH11 oncofusion 
            protein.
  JOURNAL   J Clin Invest. 2023 Dec 7:e176311. doi: 10.1172/JCI176311.
  PUBMED    38061017
REFERENCE   2  (bases 1 to 7479)
  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: "J Clin 
            Invest. 2023 Dec 7:e176311. doi: 10.1172/JCI176311."
COMMENT     Sequence Label: MCS-TurboID MSCV-IRES-GFP
FEATURES             Location/Qualifiers
     source          1..7479
                     /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"
     primer_bind     1372..1388
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     CDS             1504..2463
                     /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    2471..3038
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     misc_feature    2488..3040
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(2632..2649)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     2859..2878
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             join(3052..3054,3055..3057,3058..3771)
                     /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(3358..3377)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3705..3726
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     LTR             3873..4387
                     /label=3' LTR
                     /note="3' long terminal repeat from murine embryonic stem 
                     cell virus"
     protein_bind    4549..4565
                     /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(4573..4579,4580..4597,4598..4603))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4618..4639
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4756..4773)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4927..5515)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5007..5026)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5686..6477,6478..6546))
                     /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(6547..6651)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6719..6737
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(6775..6797)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6897..6916
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     7110..7132
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     7260..7279
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     LTR             join(7479,1..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 aggaattccc gcggatgcat
     1441 gttaacaccg gtggatccgg tggaggtggt tctggcggag gcggctctgg aggaggagga
     1501 agtatgaaag acaatactgt gcctctgaag ctgatcgctc tcctggctaa tggcgagttc
     1561 catagtggcg aacagctggg agaaaccctg ggcatgtcca gggccgctat caacaagcac
     1621 attcagactc tgcgcgactg gggcgtggac gtgttcaccg tgcccggaaa gggctactct
     1681 ctgcccgagc ctatcccgct gctgaacgct aaacagattc tgggacagct ggacggcggg
     1741 agcgtggcag tcctgcctgt ggtcgactcc accaatcagt acctgctgga tcgaatcggc
     1801 gagctgaaga gtggggatgc ttgcattgca gaatatcagc aggcagggag aggaagcaga
     1861 gggaggaaat ggttctctcc ttttggagct aacctgtacc tgagtatgtt ttggcgcctg
     1921 aagcggggac cagcagcaat cggcctgggc ccggtcatcg gaattgtcat ggcagaagcg
     1981 ctgcgaaagc tgggagcaga caaggtgcga gtcaaatggc ccaatgacct gtatctgcag
     2041 gatagaaagc tggcaggcat cctggtggag ctggccggaa taacaggcga tgctgcacag
     2101 atcgtcattg gcgccgggat taacgtggct atgaggcgcg tggaggaaag cgtggtcaat
     2161 cagggctgga tcacactgca ggaagcaggg attaacctgg acaggaatac tctggccgct
     2221 acgctgatcc gagagctgcg ggcagccctg gaactgttcg agcaggaagg cctggctcca
     2281 tatctgccac ggtgggagaa gctggataac ttcatcaata gacccgtgaa gctgatcatt
     2341 ggggacaaag agattttcgg gattagccgg gggattgata aacagggagc cctgctgctg
     2401 gaacaggacg gagttatcaa accctggatg ggcggagaaa tcagtctgcg gtctgccgaa
     2461 aagtagcaat tccgcccccc ccccctaacg ttactggccg aagccgcttg gaataaggcc
     2521 ggtgtgcgtt tgtctatatg ttattttcca ccatattgcc gtcttttggc aatgtgaggg
     2581 cccggaaacc tggccctgtc ttcttgacga gcattcctag gggtctttcc cctctcgcca
     2641 aaggaatgca aggtctgttg aatgtcgtga aggaagcagt tcctctggaa gcttcttgaa
     2701 gacaaacaac gtctgtagcg accctttgca ggcagcggaa ccccccacct ggcgacaggt
     2761 gcctctgcgg ccaaaagcca cgtgtataag atacacctgc aaaggcggca caaccccagt
     2821 gccacgttgt gagttggata gttgtggaaa gagtcaaatg gctctcctca agcgtattca
     2881 acaaggggct gaaggatgcc cagaaggtac cccattgtat gggatctgat ctggggcctc
     2941 ggtgcacatg ctttacatgt gtttagtcga ggttaaaaaa acgtctaggc cccccgaacc
     3001 acggggacgt ggttttcctt tgaaaaacac gatgataata tggccacaac catggtgagc
     3061 aagggcgagg agctgttcac cggggtggtg cccatcctgg tcgagctgga cggcgacgta
     3121 aacggccaca agttcagcgt gtctggcgag ggcgagggcg atgccaccta cggcaagctg
     3181 accctgaagt tcatctgcac caccggcaag ctgcccgtgc cctggcccac cctcgtgacc
     3241 accctgacct acggcgtgca gtgcttcagc cgctaccccg accacatgaa gcagcacgac
     3301 ttcttcaagt ccgccatgcc cgaaggctac gtccaggagc gcaccatctt cttcaaggac
     3361 gacggcaact acaagacccg cgccgaggtg aagttcgagg gcgacaccct ggtgaaccgc
     3421 atcgagctga agggcatcga cttcaaggag gacggcaaca tcctggggca caagctggag
     3481 tacaactaca acagccacaa cgtctatatc atggccgaca agcagaagaa cggcatcaag
     3541 gcgaacttca agatccgcca caacatcgag gacggcagcg tgcagctcgc cgaccactac
     3601 cagcagaaca cccccatcgg cgacggcccc gtgctgctgc ccgacaacca ctacctgagc
     3661 acccagtccg ccctgagcaa agaccccaac gagaagcgcg atcacatggt cctgctggag
     3721 ttcgtgaccg ccgccgggat cactctcggc atggacgagc tgtacaagta atgaattaat
     3781 taagaattat caagcttatc gataccgtcg acctgcagcc aagcttatcg ataaaataaa
     3841 agattttatt tagtctccag aaaaaggggg gaatgaaaga ccccacctgt aggtttggca
     3901 agctagctta agtaacgcca ttttgcaagg catggaaaat acataactga gaatagagaa
     3961 gttcagatca aggttaggaa cagagagaca gcagaatatg ggccaaacag gatatctgtg
     4021 gtaagcagtt cctgccccgg ctcagggcca agaacagatg gtccccagat gcggtcccgc
     4081 cctcagcagt ttctagagaa ccatcagatg tttccagggt gccccaagga cctgaaatga
     4141 ccctgtgcct tatttgaact aaccaatcag ttcgcttctc gcttctgttc gcgcgcttct
     4201 gctccccgag ctcaataaaa gagcccacaa cccctcactc ggcgcgccag tcctccgata
     4261 gactgcgtcg cccgggtacc cgtgtatcca ataaaccctc ttgcagttgc atccgacttg
     4321 tggtctcgct gttccttggg agggtctcct ctgagtgatt gactacccgt cagcgggggt
     4381 ctttcatggg taacagtttc ttgaagttgg agaacaacat tctgagggta ggagtcgaat
     4441 attaagtaat cctgactcaa ttagccactg ttttgaatcc acatactcca atactcctga
     4501 aatagttcat tatggacagc gcagaaagag ctggggagaa ttgtgaaatt gttatccgct
     4561 cacaattcca cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg
     4621 agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct
     4681 gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg
     4741 gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc
     4801 ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg
     4861 aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct
     4921 ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca
     4981 gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct
     5041 cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc
     5101 gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt
     5161 tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc
     5221 cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc
     5281 cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg
     5341 gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc
     5401 agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag
     5461 cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga
     5521 tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat
     5581 tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag
     5641 ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat
     5701 cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc
     5761 cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat
     5821 accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag
     5881 ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg
     5941 ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc
     6001 tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca
     6061 acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg
     6121 tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc
     6181 actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta
     6241 ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc
     6301 aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg
     6361 ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc
     6421 cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc
     6481 aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat
     6541 actcatactc ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag
     6601 cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc
     6661 ccgaaaagtg ccacctgacg tctaagaaac cattattatc atgacattaa cctataaaaa
     6721 taggcgtatc acgaggccct ttcgtctcgc gcgtttcggt gatgacggtg aaaacctctg
     6781 acacatgcag ctcccggaga cggtcacagc ttgtctgtaa gcggatgccg ggagcagaca
     6841 agcccgtcag ggcgcgtcag cgggtgttgg cgggtgtcgg ggctggctta actatgcggc
     6901 atcagagcag attgtactga gagtgcacca tatgcggtgt gaaataccgc acagatgcgt
     6961 aaggagaaaa taccgcatca ggcgccattc gccattcagg ctgcgcaact gttgggaagg
     7021 gcgatcggtg cgggcctctt cgctattacg ccagctggcg aaagggggat gtgctgcaag
     7081 gcgattaagt tgggtaacgc cagggttttc ccagtcacga cgttgtaaaa cgacggcgca
     7141 aggaatggtg catgcaagga gatggcgccc aacagtcccc cggccacggg gcctgccacc
     7201 atacccacgc cgaaacaagc gctcatgagc ccgaagtggc gagcccgatc ttccccatcg
     7261 gtgatgtcgg cgatataggc gccagcaacc gcacctgtgg cgccggtgat gccggccacg
     7321 atgcgtccgg cgtagaggcg attagtccaa tttgttaaag acaggatatc agtggtccag
     7381 gctctagttt tgactcaaca atatcaccag ctgaagccta tagagtacga gccatagata
     7441 aaataaaaga ttttatttag tctccagaaa aagggggga
//
